To evaluate the apoptosis positivity, the expression of Bcl-2, bax proteinsin 30 patients with squamous cell cervix carcinoma before and after radiotherapy. Methods: By usingimmuno-histochemical and TDT-dUTP nick end ...To evaluate the apoptosis positivity, the expression of Bcl-2, bax proteinsin 30 patients with squamous cell cervix carcinoma before and after radiotherapy. Methods: By usingimmuno-histochemical and TDT-dUTP nick end labelling techniques, 30 cases of squamous cell cervicalcarcinoma were analyzed. Results: The apoptosis positivity before and after irradiation was 76.7%and 100% respectively, with the difference being significant (P 【 0.05); The positive rates of Bcl-2protein before and after irradiation were 73.3% and 46.7% respectively, with the difference beingsignificant (P 【 0.05); The positive rates of bax protein before and after irradiation were 86% and100% respectively, with the difference being significant (P 【 0.05). Conclusion: bax and Bcl-2protein play an important role in apoptosis induced by fractionated radiation therapy. Apoptosisinduced by irradiation is contributed to upregulation of bax protein or downregulation of Bcl-2protein.展开更多
Objective: To investigate the expressions of cyclooxygenase (COX)-2 and matrix metalloproteinase (MMP)-9 in cervical carcinoma and their clinical significance. Methods: Immunohistochemistry SP method was used to detec...Objective: To investigate the expressions of cyclooxygenase (COX)-2 and matrix metalloproteinase (MMP)-9 in cervical carcinoma and their clinical significance. Methods: Immunohistochemistry SP method was used to detect the expres- sions of COX-2 and MMP-9 in 72 cases of invasive carcinoma of cervix (ICC) and 16 cases of normal cervical epithelium remote from tumor (NCE). The relationships between the expressions of COX-2, MMP-9 in ICC and some characteristics relating to clinical pathology of cervical carcinoma such as histological grading, lymph node metastasis, stromal invasion and FIGO stage were analyzed statistically. Results: The rates of the positive expressions of COX-2 and MMP-9 in ICC were significantly higher than those in NCE. COX-2: 88.9% (64/72) in group ICC and 12.5% (2/16) in group NCE, P = 0.000; MMP-9: 94.4% (68/72) in group ICC and 43.8% (7/16) in group NCE, P = 0.000. The expression of COX-2 was positively correlated with lymph node metastasis (r = 0.296, P = 0.012) and stromal invasion (r = 0.257, P = 0.029). The expression of MMP-9 was positively correlated with FIGO stage (r = 0.329, P = 0.005) and histological grading (r = 0.351, P = 0.003). The expression of COX-2 was positively correlated with the expression of MMP-9 in ICC (r = 0.297, P = 0.011). Conclusion: The overexpressions of COX-2 and MMP-9 were closely related to the invasion and growth of cervical carcinoma. The tissue with the overexpression of COX-2 had strong invasion ability. COX-2 and MMP-9 had synergistic effect on proliferation, invasion and metastasis of cancer cells. Detecting the coexpression of COX-2 and MMP-9 may be of value in further understanding the biological behavior and predicting the prognosis of cervical carcinoma.展开更多
Objective: The purpose of the study is to investigate the effects of up-regulation of Raf kinase inhibitor protein (RKIP) on the chemosensitivity of cervical cancer Hela cells. Methods: Eukaryotic expression plasm...Objective: The purpose of the study is to investigate the effects of up-regulation of Raf kinase inhibitor protein (RKIP) on the chemosensitivity of cervical cancer Hela cells. Methods: Eukaryotic expression plasmid pcDNA3.1(±)-ssRKIP containing human overall length RKIPcDNA was transfected into cervical cancer Hela cell by lipofectin assay, establishing a stable cell line containing a target gene by G418. Expression of RKIP in Hela cells was measured by Western blot analysis. After treatment with cisplatin of different concentrations and intervals of time, the effect of RKIP on the proliferation of Hela cells was evaluated by MTT method. The flow cytometry was used to investigate whether the RKIP could inhibit apoptosis in Hela cells induced by cisplatin. Results: The expression of RKIP in Hela cells transfected with pcDNA3.1-ssRKIP was increased obviously. After different concentrations of cisplatin treatment cells for 24, 48 and 72 h, the growth inhibition rate in Hela cells transfected with pcDNA3.1-ssRKIP was significantly higher than in control cells (P 〈 0.05). With 5 pg/mL cisplatin treatment for 24 h, pcDNA3.1-ssRKIP-transfected Hela cells had an obviously higher percentage of apoptosis (23.2 ± 0.24)% than non-transfected cells (12.4 ± 0.31)% and empty vector-transfected cells (13.4 ± 0.47)%. Without treatment of cisplatin, the percentage of apoptosis for Hela cells transfected with pcDNA3.1-ssRKIP was (5.7 ± 0.12)%, which was still higher than those of the non-transfected cells (2.9 ± 0.21)% and empty vector-transfected cells (3 ± 0.08)%. Conclusion: Higher expres- sion of RKIP gene can improve chemosensitivitv of cervical cancer Hela cells to cisplatin.展开更多
In searching of differentially expressed genes in human uterine leiomyomas, differential display was used with twelve pairs of primers to compare human uterine leiomyomas with matched myometrium. False positives were ...In searching of differentially expressed genes in human uterine leiomyomas, differential display was used with twelve pairs of primers to compare human uterine leiomyomas with matched myometrium. False positives were eliminated by reverse Northern analysis. Positives were confirmed by Northern blot analysis. RESULTS: Four of 69 cDNA fragments (3 up-regulated named L1, L2 and L3 and 1 down-regulated named M1 in leiomyoma) were confirmed by Northern analysis. Sequence comparison and Northern analysis proved that L1 is exactly the human ribosomal protein S19. It was present ubiquitously in 13 tissues tested but in various levels and even in different size. L1 was highly expressed in parotidean cystadenocarcinoma, pancreatic cancer and breast cancer examined. No mutations have been found in human uterine leiomyomas (n=6). CONCLUSIONS: hRPS19 overexpression might be a universal signal in rapid cell growth tissues.展开更多
The aim of this study is to investigate cyclin E, pl6ink4a and ki67 as possible diagnostic biomarkers for cervical preneoplasia using cervical exfoliated-cell specimens, and evaluate the significance for screening pat...The aim of this study is to investigate cyclin E, pl6ink4a and ki67 as possible diagnostic biomarkers for cervical preneoplasia using cervical exfoliated-cell specimens, and evaluate the significance for screening patients at high risk of de- veloping cervical carcinoma. The expression of cyclin E, p16ink4a and ki67 was examinated in 78 cervical exfoliated epithe- lial specimens diagnosed as atypical squamous cells of undetermined significance (ASCIIS) (12 cases), cervical intraepithe- lial neoplasia (CIN) of type 1 (17 cases), CIN2-3(38 cases) and invasive carcinoma (11 cases) using immunohistochemical analysis, and simultaneously, the DNA status of human papillomavirus (HPV) type 16/18 was detected by polymerase chain reaction (PCR) using type specific primers, cyclin E, pl6ink4a and ki67 were all overexpressed in CINs and invasive carci- noma, compared with little expression in ASCUS ( P < 0.005). Overexpression of cyclin E was observed in CIN1 (94.1 % , x2 = 21.16, P < 0.01), and pl6ink4a and ki67 were overexpressed in invasive carcinoma (100% and 90.9% respective- ly) . The degree of pl6ink4a and ki67 expression correlated well with that of epithelial lesions ( P < 0.005). HPV16/18 infec- tion was assessed in CINs and invasive carcinoma samples, and revealed a significant relationship with the degree of cervical epithelial lession. The expression level of pl6ink4a and ki67 seemed more closely associated with HPV16 infection than that of cyclin E ( r s = 1.0 vs rs = 0 . 4 ) . Only 1 case in CIN, and 4 cases in CIN2-3 of HPV18 positive samples were detected. Therefore no statistical significance was found by statistical analysis. Overexpression of cyclin E, p16ink4a and ki67 in CINs and invasive carcinoma cells demonstrates the potential use of cyclin E, p16ink4a and ki67 as diagnostic biomarkers for HPV-related cervical neoplastic lesions. In addition, this technique can be used for screening patients at high risk of devel- oping cervical carcinoma.展开更多
To clone and identify the proteins involved in regulating the transcription of hTERT and study the role of genes in both hTERT transcription and telomerase activity. Methods The full cDNA of COUP-TFII was clon...To clone and identify the proteins involved in regulating the transcription of hTERT and study the role of genes in both hTERT transcription and telomerase activity. Methods The full cDNA of COUP-TFII was cloned from HeLa cDNA library by hTERT promoter-based yeast one-hybrid assay and then in-frame inserted into His-tag fusion expression vector pEK318. The His-tag COUP-TFII fusion proteins were purified by Ni-NTA chromatography. The interaction of COUP-TFII with hTERT promoter in vitro was identified by electrophoretic mobility shift assay and Footprint. The role of COUP-TFII in both hTERT transcription and telomerase activity were probed through Luciferase reporter assay, Northern blot, and TRAP-PCR ELISA. Results COUP-TFII could firmly bind to the downstream E-box and the other two binding sites in hTERT promoter. Luciferase reporter assay indicated COUP-TFII could suppress hTERT promoter activity and stable introduction of COUP-TFII into HeLa cells also decreased both endogenous hTERT transcription and telomerase activity. Conclusion The human COUP-TFII can firmly bind to hTERT promoter, and inhibit telomerase activity through decreasing hTERT transcription. It will greatly facilitate understanding of telomerase regulation in normal and cancer cells展开更多
Objective: To investigate the effects of lithium on cognitive function and metabolism of Amyloid-beta Protein Precursor (APP) and tau phosphorylation in rats chronically exposed to aluminum. Methods: Twenty-four chron...Objective: To investigate the effects of lithium on cognitive function and metabolism of Amyloid-beta Protein Precursor (APP) and tau phosphorylation in rats chronically exposed to aluminum. Methods: Twenty-four chronically aluminum-exposed rats were randomly divided into 2 groups: a lithium-treatment group and a non-treatment group (n=12 per group). Lithium chloride was administered to the lithium-treatment group via gastric gavage daily for 6 weeks (200 mg/kg·d LiCl), while the non-treatment group was administered the same volume of sodium chloride by the same means. An additional control group (n=12) received no intervention. Memory function was evaluated by the Morris water maze test. Aβ was measured by immunohistochemical staining, while total APP, phosphorylated-tau protein, CDK5 and PP2A were determined by Western Blotting. Results: (1) Compared to the non-treatment group, the lithium-treatment group had a significantly shorter mean escape latency and a lower proportion of random navigation pattern in the spatial probe test (P<0.05). After the platform was taken away, the rats in the lithium-treatment group crossed the platform quadrant significantly more and stayed longer in the platform quadrant than those in the non-treatment group (P<0.05). (2) The number of Aβ positive neurons in the hippocampus and cortex was significantly less in the lithium-treatment group than in the non-treatment group (P<0.05), but the content of APP was not different between groups (P=0.730). (3) Phosphorylation of tau protein decreased significantly in the lithium-treatment group than that in the non-treatment group (P<0.05). The content of CDK5 in the lithium-treatment group was significantly less than that in the non-treatment group in the cortex and hippocampus, while there was no difference in the content of PP2A between the 2 groups. The expression of CDK5 was significantly correlated with phosphorylated tau (r=0.871, P=0.024) in the lithium-treatment group. Conclusion: Lithium may improve memory function in rats chronically exposed to aluminum by decreasing both the production of Aβ and tau phosphorylation, with the latter results from inhibiting expression of CDK5.展开更多
Objective: To investigate the proteornic differences between the high-sensitivity (HS) group and low-sensitivity (LS) group of cervical cancer treated by radiotherapy and confirm the radiotherapy sensitivity asso...Objective: To investigate the proteornic differences between the high-sensitivity (HS) group and low-sensitivity (LS) group of cervical cancer treated by radiotherapy and confirm the radiotherapy sensitivity associated proteins in early cervical cancer. Methods: The fresh carcinoma tissues were collected from 10 untreated cervical cancer patients and preserved in the -80 ℃ refrigeratory. The tissues were classified into two groups: high sensitivity group (HS) and low sensitivity group (LS), according to their response to radiotherapy. In the first part of our experiment, protein separating was performed by using two-dimensional gel electrophoresis (2-DE) with Amersham 18 cm linear pH 3-10 immobilized pH gradient (IPG) strips. The images of the gels were acquired by the scanner and then analyzed by using PD-quest7.3 software to find the differentially expression protein-spots in each group. Then the differentially expressed protein-spots was incised from the gels and digested by trypsin. The peptide mass fingerprintings (PMF) was acquired by matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and the proteins were identified by data searching in the Mascot-database. Part of differentially expression proteins were assayed by Western Blot. Results: Most of the gels were clear and successfully analyzed by PD-quest7.3 software. Most of the protein-spots concentrated on the area of 20-100Kda (Mw) and pH4-8. The average number of the protein-spots was 754 ± 64 in HS group and 777 ±48个 in LS group. The match rate was 87.6% between two groups. Five high expression proteins were found in HS group which were low expression in LS group, 3 high expression protein were found in LS group which were low expression in HS group. Reselts of Western Blot were in coincidence to proteomic result. Conclusion: The 2-DE gels image of HS group and LS group with early cervical cancer tissues treated by radiotherapy are successfully acquired. Some differentially expression proteins between the two groups are further confirmed by immunohistochemical assay.展开更多
Objective This study aimed to combine tumor abnormal protein(TAP) and high-sensitivity C-reactive protein(hs-CRP) level detection to diagnose endometrial cancer in patients with endometrial thickness less than 8 mm, a...Objective This study aimed to combine tumor abnormal protein(TAP) and high-sensitivity C-reactive protein(hs-CRP) level detection to diagnose endometrial cancer in patients with endometrial thickness less than 8 mm, and to provide a reference for clinical screening and diagnosis. Methods Clinical data from 19 cases of endometrial cancer, diagnosed on the basis of pathological findings, were collected from September 2014 to December 2015. The inclusion criteria were as follows: the patients were first diagnosed with endometrial thickness less than 8 mm and were all in menopause. Perimenopausal patients(n = 26) with uterine fibroids seen during the same period were selected as a control group. Serum TAP and hs-CRP levels of the patients in the two groups were simultaneously determined on admission. Results We found that both TAP and hs-CRP levels in the experimental group were higher than those in the control group [(182.95 ± 72.14) μm^2 vs.(133.19 ± 55.18) μm^2, P = 0.019;(7.52 ± 19.03) mg/L vs.(1.66 ± 2.31) mg/L, P = 0.136]. The sensitivity of TAP for the diagnosis of endometrial cancer was 73.68%, the specificity was 69.23%, and the Youden index was 0.4291. The diagnostic sensitivity and specificity of hs-CRP was 15.79% and 100%, respectively, and the Youden index was 0.1579. After plotting the receiver operating characteristics curves, the optimal cut-off value for TAP in diagnosing endometrial cancer was found to be 160.662 μm^2 and that for hs-CRP was 1.07 mg/L. Conclusion For patients suspected of having endometrial cancer with endometrial thickness less than 8 mm, combined detection of TAP and hs-CRP levels can be used as a screening tool and can provide new ideas regarding clinical diagnosis and treatment.展开更多
Objective: To retrospectively analyze the clinical data of 275 cases of cervical cancer (CC) in our hospital, and investigate the clinicopathological parameters of cervical cancer based on the expression of p16INK4A p...Objective: To retrospectively analyze the clinical data of 275 cases of cervical cancer (CC) in our hospital, and investigate the clinicopathological parameters of cervical cancer based on the expression of p16INK4A proteins. Methods: The clinical information of 275 patients with cervical cancer were retrospectively analyzed between 2006 and 2011, including the patients' age, clinical FIGO stage, differentiation, histologic grade, infiltration depth, treatment, pathological diagnosis after surgery, and results of following-up.Immunohistochemistry was also done on sections of confirmed cancer specimens without prior chemotherapy/radiotherapy. Results: Among various clinicopathological parameters, the median age was 48 years old. The grade was significantly associated with histological type, HPV infection and with lymph node invasion. FIGO stage was strongly correlated to the infiltration depth and lymph node metastasis. P16INK4A expression was significantly correlated with histologic grade. However, there were no differences between p16INK4A staining and patient's age, histopathology and lymph node metastases. Conclusion: The incidence of cervical cancer becomes increasingly younger. Additionally, p16INK4A can function as a diagnostic marker of cervical carcinomas.展开更多
To investigate the effects of early nutritional intervention on the serum insulin-like growth factor-1 (IGF1),insulin-like growth factor binding protein 3 (IGFBP3), intestinal development, and catch-up growth of intra...To investigate the effects of early nutritional intervention on the serum insulin-like growth factor-1 (IGF1),insulin-like growth factor binding protein 3 (IGFBP3), intestinal development, and catch-up growth of intrauterine growth retardation (IUGR) rats by giving the IUGR new born rats different protein level diet. Methods IUGR rat model was built by starvation of pregnant female rats. Twenty-four IUGR pups and 8 normal pups were divided randomly into 4 groups: normal control group (C group); IUGR control group(S group), IUGR low-protein diet group (SL group), and IUGR high-protein diet group (SH group). Detected the serum IGF1, IGFBP3, body weight, body length, intestinal weight length, intestinal villi height (VH), crypt depth (CD), villi absorbing area (VSA), mucous thickness (MT), and disaccharidase at the 4th week. Results (1) The SH group showed the fastest catch-up growth, serum IGF1, IGFBP3, VH, and VSA were significantly higher than those of normal control group and IUGR control group. The intestinal weight and length, and the activities of lactase and saccharase of the SH group also reached the normal control group level. (2) The SL group kept on small size, the serum IGF1, IGFBP3, and most of intestinal histological indexes were all significantly lower than other groups. (3) IGF-1, IGFBP3 were positively correlated to intestinal VH, VSA, saccharase, body weight and length. Conclusions The serum IGF1 was a sensitive index to the catch-up growth. The early nutritional intervention of high-protein diet after birth is helpful for the catch-up growth of IUGR through promoting the intestinal development and the ab-sorption of nutrition展开更多
The objective was to investigate the expression of bone morphogenetic protein (BMP) family members in the mouse uterus during the estrous cycle by real-time polymerase chain reaction (PCR) and immunohistochemistry...The objective was to investigate the expression of bone morphogenetic protein (BMP) family members in the mouse uterus during the estrous cycle by real-time polymerase chain reaction (PCR) and immunohistochemistry. Uterine samples from Swiss ICR mice were collected and dissected free of surrounding tissue. One uterine horn was snap frozen in liquid nitrogen immediately after collection and stored at -80 ℃for RNA extraction, and the other was fixed in 40 mg/ml paraformaldehyde at room temperature for immunolocalization of BMP2 protein. Real-time PCR analysis showed that the expression level of Bmp2 was significantly higher at proestrus than at estrus and metestrus (P〈0.05). The relative abundance of Bmp4 exhibited significant fluctuations, but there were no statistically significant differences between the expression levels of Bmp2 and Bmp4 (P〉0.05). The expression levels of Bmprla and Bmpr2 remained unchanged during estrous cycles. However, the level of Bmprlb mRNA decreased significantly at estrus (P〈0.05), increasing subsequently at metestrus. Furthermore, the level of Bmprlb mRNA was significantly lower than those of Bmprla and Bmpr2 mRNA at the corresponding stages (P〈0.05). All three receptor-regulated Smads (R-Smads) detected were differentially expressed in the mouse uterus and the expression levels of Smadl and Smad5 were significantly higher than that of Smad8 (P〈0.05). In addition, the expression level of Smad4 did not change substantially throughout the estrous cycle. Immunohistochemical experiments revealed that BMP2 protein was differentially expressed and localized mainly in the uterine luminal and glandular epithelial cells throughout the estrous cycle. In conclusion, our results provide information about the variation in the mRNA levels of Bmp2 and Bmp4 and related components of the BMP signaling pathway. The data provide quantitative and useful information about the roles of endometrial BMP proposed and demonstrated by others, such as the degradation and remodeling of the endometrium.展开更多
OBJECTIVE:To observe the effects of Compound Danshen Tablets(CDST) on spatial cognition and expression of brain b-amyloid precursor protein(β-APP) in a rat model of Alzheimer's disease.METHODS:The rat model of Al...OBJECTIVE:To observe the effects of Compound Danshen Tablets(CDST) on spatial cognition and expression of brain b-amyloid precursor protein(β-APP) in a rat model of Alzheimer's disease.METHODS:The rat model of Alzheimer's disease(AD) was established using D-galactose to cause subacute aging combined with Meynert nucleus damage.Rat behavior was monitored using the Morris water maze,and the expression of β-APP in rat brain tissue was detected via immunohistochemistry.RESULTS:CDST significantly improved spatial cognition and decreased β-APP expression in the cortex and hippocampus(P<0.05,P<0.01).CONCLUSIONS:CDST can significantly improve spatial cognition in a rat model of AD.This observation is possibly related to a reduction in β-APP ex-pression in the rat brain.展开更多
Objective It has been shown that there are extensive interactions between the central nervous system and the immune system.The present study focused on the effects of lipopolysaccharide(LPS)on memory retrieval,to ex...Objective It has been shown that there are extensive interactions between the central nervous system and the immune system.The present study focused on the effects of lipopolysaccharide(LPS)on memory retrieval,to explore the interaction between immune activation and memory.Methods C57BL/6J mice(8 weeks old)were first trained in the Morris water maze to reach asymptotic performance.Then mice were tested 24 h after the last training session and LPS was administered(1.25 mg/kg,i.p.)4 h prior to the testing.The retrieval of spatial memory was tested by probe trial,and the time spent in the target quadrant and the number of platform location crosses were recorded.ELISA was performed to detect interleukin-1β(IL-1β)protein level in the hippocampus of mice tested in the water maze.Results Although LPS induced overt sickness behavior and a significant increase in the level of IL-1β in the hippocampus of mice,there was no significant difference in the time spent in the target quadrant or in the number of platform location crosses between LPS-treated and control groups in the probe trial testing.Conclusion Immune activation induced by LPS does not impair the retrieval of spatial memory.展开更多
Maternal stress during pregnancy is prevailing worldwide, which exposes fetuses to intrauterine hyper glucocorticoids(GC), programming offspring to obesity and metabolic diseases. Despite the importance of brown adipo...Maternal stress during pregnancy is prevailing worldwide, which exposes fetuses to intrauterine hyper glucocorticoids(GC), programming offspring to obesity and metabolic diseases. Despite the importance of brown adipose tissue(BAT) in maintaining long-term metabolic health, impacts of prenatal hyper GC on postnatal BAT thermogenesis and underlying regulations remain poorly defined. Pregnant mice were administrated with synthetic GC dexamethasone(DEX) at levels comparable to fetal GC exposure of stressed mothers. Prenatal GC exposure dose-dependently reduced BAT thermogenic activity, contributing to lower body temperature and higher mortality of neonates;such difference was abolished under thermoneutrality, underscoring BAT deficiency was the major contributor to adverse changes in postnatal thermogenesis due to excessive GC. Prenatal GC exposure highly activated Redd1 expression and reduced Ppargc1 a transcription from the alternative promoter(Ppargc1 a-AP) in neonatal BAT. During brown adipocyte differentiation, ectopic Redd1 expression reduced Ppargc1 a-AP expression and mitochondrial biogenesis;and the inhibitory effects of GC on mitochondrial biogenesis and Ppargc1 a-AP expression were blocked by Redd1 ablation. Redd1 reduced protein kinase A phosphorylation and suppressed cyclic adenosine monophosphate(c AMP)-responsive element-binding protein(CREB) binding to the c AMP regulatory element(CRE) in Ppargc1 a-AP promoter, leading to Ppargc1 a-AP inactivation. In summary, excessive maternal GC exposure during pregnancy dysregulates Redd1-Ppargc1 a-AP axis, which impairs fetal BAT development, hampering postnatal thermogenic adaptation and metabolic health of offspring.展开更多
OBJECTIVE:To investigate the effect of Chinese medicines using the warming Yang and removing blood stasis method on levels of matrix metalloproteinases(MMPs)/tissue inhibitor metalloproteinases(TIMPs) secreted by cult...OBJECTIVE:To investigate the effect of Chinese medicines using the warming Yang and removing blood stasis method on levels of matrix metalloproteinases(MMPs)/tissue inhibitor metalloproteinases(TIMPs) secreted by cultured endometrial cells from patients with endometriosis.METHODS:Ectopic and eutopic endometrial cells obtaind from 15 endometriosis patients were cultured in vitro,and divided randomly into five groups:high dose;moderate dose;low dose;nemestran;blank control.The three dose groups were treated with a decoction prepared according to the principle of warming Yang and removing blood stasis;nemestran and 0.9%NaCI were administered to the nemestran group and balnk control group,respectively.Eutopic endometrial cells obtaind from 10 hysteromyoma patients were cultured in vitro,as the normal control group,0.9%NaCI were administered to the normal control group.Cell culture supernatants were collected and levels of matrix metalloproteinase-1(MMP-1),matrix metalloproteinase-2(MMP-2),matrix metalloproteinase-9(MMP-9),tissue inhibitor metalloproteinase-1(TIMP-1) and tissue inhibitor metalloproteinase-2(TIMP-2) detected by enzyme-linked immuno sorbent assay(ELISA).RESULTS:Compared with the normal control group,levels of MMP-1,MMP-2,and MMP-9 in eutopic and ectopic endometrium cell supernatants in the blank control group were increased,whereas levels of TIMP-1 and TIMP-2 were decreased(P <0.05).Compared with the blank control group,levels of MMP-1 and MMP-2 in ectopic and eutopic endometrium cell supernatants cultured in low-dose,middle-dose,and high-dose groups were decreased,whereas levels of TIMP-1 and TIMP-2 were increased significantly(P < 0.05).CONCLUSION:The warming Yang and removing blood stasis method affects expression of MMPs andTIMPs.展开更多
文摘To evaluate the apoptosis positivity, the expression of Bcl-2, bax proteinsin 30 patients with squamous cell cervix carcinoma before and after radiotherapy. Methods: By usingimmuno-histochemical and TDT-dUTP nick end labelling techniques, 30 cases of squamous cell cervicalcarcinoma were analyzed. Results: The apoptosis positivity before and after irradiation was 76.7%and 100% respectively, with the difference being significant (P 【 0.05); The positive rates of Bcl-2protein before and after irradiation were 73.3% and 46.7% respectively, with the difference beingsignificant (P 【 0.05); The positive rates of bax protein before and after irradiation were 86% and100% respectively, with the difference being significant (P 【 0.05). Conclusion: bax and Bcl-2protein play an important role in apoptosis induced by fractionated radiation therapy. Apoptosisinduced by irradiation is contributed to upregulation of bax protein or downregulation of Bcl-2protein.
文摘Objective: To investigate the expressions of cyclooxygenase (COX)-2 and matrix metalloproteinase (MMP)-9 in cervical carcinoma and their clinical significance. Methods: Immunohistochemistry SP method was used to detect the expres- sions of COX-2 and MMP-9 in 72 cases of invasive carcinoma of cervix (ICC) and 16 cases of normal cervical epithelium remote from tumor (NCE). The relationships between the expressions of COX-2, MMP-9 in ICC and some characteristics relating to clinical pathology of cervical carcinoma such as histological grading, lymph node metastasis, stromal invasion and FIGO stage were analyzed statistically. Results: The rates of the positive expressions of COX-2 and MMP-9 in ICC were significantly higher than those in NCE. COX-2: 88.9% (64/72) in group ICC and 12.5% (2/16) in group NCE, P = 0.000; MMP-9: 94.4% (68/72) in group ICC and 43.8% (7/16) in group NCE, P = 0.000. The expression of COX-2 was positively correlated with lymph node metastasis (r = 0.296, P = 0.012) and stromal invasion (r = 0.257, P = 0.029). The expression of MMP-9 was positively correlated with FIGO stage (r = 0.329, P = 0.005) and histological grading (r = 0.351, P = 0.003). The expression of COX-2 was positively correlated with the expression of MMP-9 in ICC (r = 0.297, P = 0.011). Conclusion: The overexpressions of COX-2 and MMP-9 were closely related to the invasion and growth of cervical carcinoma. The tissue with the overexpression of COX-2 had strong invasion ability. COX-2 and MMP-9 had synergistic effect on proliferation, invasion and metastasis of cancer cells. Detecting the coexpression of COX-2 and MMP-9 may be of value in further understanding the biological behavior and predicting the prognosis of cervical carcinoma.
基金Supported by a grant from the Qingdao Public Sphere Sci-technical Support Project(No.09-1-1-13-nsh)
文摘Objective: The purpose of the study is to investigate the effects of up-regulation of Raf kinase inhibitor protein (RKIP) on the chemosensitivity of cervical cancer Hela cells. Methods: Eukaryotic expression plasmid pcDNA3.1(±)-ssRKIP containing human overall length RKIPcDNA was transfected into cervical cancer Hela cell by lipofectin assay, establishing a stable cell line containing a target gene by G418. Expression of RKIP in Hela cells was measured by Western blot analysis. After treatment with cisplatin of different concentrations and intervals of time, the effect of RKIP on the proliferation of Hela cells was evaluated by MTT method. The flow cytometry was used to investigate whether the RKIP could inhibit apoptosis in Hela cells induced by cisplatin. Results: The expression of RKIP in Hela cells transfected with pcDNA3.1-ssRKIP was increased obviously. After different concentrations of cisplatin treatment cells for 24, 48 and 72 h, the growth inhibition rate in Hela cells transfected with pcDNA3.1-ssRKIP was significantly higher than in control cells (P 〈 0.05). With 5 pg/mL cisplatin treatment for 24 h, pcDNA3.1-ssRKIP-transfected Hela cells had an obviously higher percentage of apoptosis (23.2 ± 0.24)% than non-transfected cells (12.4 ± 0.31)% and empty vector-transfected cells (13.4 ± 0.47)%. Without treatment of cisplatin, the percentage of apoptosis for Hela cells transfected with pcDNA3.1-ssRKIP was (5.7 ± 0.12)%, which was still higher than those of the non-transfected cells (2.9 ± 0.21)% and empty vector-transfected cells (3 ± 0.08)%. Conclusion: Higher expres- sion of RKIP gene can improve chemosensitivitv of cervical cancer Hela cells to cisplatin.
文摘In searching of differentially expressed genes in human uterine leiomyomas, differential display was used with twelve pairs of primers to compare human uterine leiomyomas with matched myometrium. False positives were eliminated by reverse Northern analysis. Positives were confirmed by Northern blot analysis. RESULTS: Four of 69 cDNA fragments (3 up-regulated named L1, L2 and L3 and 1 down-regulated named M1 in leiomyoma) were confirmed by Northern analysis. Sequence comparison and Northern analysis proved that L1 is exactly the human ribosomal protein S19. It was present ubiquitously in 13 tissues tested but in various levels and even in different size. L1 was highly expressed in parotidean cystadenocarcinoma, pancreatic cancer and breast cancer examined. No mutations have been found in human uterine leiomyomas (n=6). CONCLUSIONS: hRPS19 overexpression might be a universal signal in rapid cell growth tissues.
文摘The aim of this study is to investigate cyclin E, pl6ink4a and ki67 as possible diagnostic biomarkers for cervical preneoplasia using cervical exfoliated-cell specimens, and evaluate the significance for screening patients at high risk of de- veloping cervical carcinoma. The expression of cyclin E, p16ink4a and ki67 was examinated in 78 cervical exfoliated epithe- lial specimens diagnosed as atypical squamous cells of undetermined significance (ASCIIS) (12 cases), cervical intraepithe- lial neoplasia (CIN) of type 1 (17 cases), CIN2-3(38 cases) and invasive carcinoma (11 cases) using immunohistochemical analysis, and simultaneously, the DNA status of human papillomavirus (HPV) type 16/18 was detected by polymerase chain reaction (PCR) using type specific primers, cyclin E, pl6ink4a and ki67 were all overexpressed in CINs and invasive carci- noma, compared with little expression in ASCUS ( P < 0.005). Overexpression of cyclin E was observed in CIN1 (94.1 % , x2 = 21.16, P < 0.01), and pl6ink4a and ki67 were overexpressed in invasive carcinoma (100% and 90.9% respective- ly) . The degree of pl6ink4a and ki67 expression correlated well with that of epithelial lesions ( P < 0.005). HPV16/18 infec- tion was assessed in CINs and invasive carcinoma samples, and revealed a significant relationship with the degree of cervical epithelial lession. The expression level of pl6ink4a and ki67 seemed more closely associated with HPV16 infection than that of cyclin E ( r s = 1.0 vs rs = 0 . 4 ) . Only 1 case in CIN, and 4 cases in CIN2-3 of HPV18 positive samples were detected. Therefore no statistical significance was found by statistical analysis. Overexpression of cyclin E, p16ink4a and ki67 in CINs and invasive carcinoma cells demonstrates the potential use of cyclin E, p16ink4a and ki67 as diagnostic biomarkers for HPV-related cervical neoplastic lesions. In addition, this technique can be used for screening patients at high risk of devel- oping cervical carcinoma.
文摘To clone and identify the proteins involved in regulating the transcription of hTERT and study the role of genes in both hTERT transcription and telomerase activity. Methods The full cDNA of COUP-TFII was cloned from HeLa cDNA library by hTERT promoter-based yeast one-hybrid assay and then in-frame inserted into His-tag fusion expression vector pEK318. The His-tag COUP-TFII fusion proteins were purified by Ni-NTA chromatography. The interaction of COUP-TFII with hTERT promoter in vitro was identified by electrophoretic mobility shift assay and Footprint. The role of COUP-TFII in both hTERT transcription and telomerase activity were probed through Luciferase reporter assay, Northern blot, and TRAP-PCR ELISA. Results COUP-TFII could firmly bind to the downstream E-box and the other two binding sites in hTERT promoter. Luciferase reporter assay indicated COUP-TFII could suppress hTERT promoter activity and stable introduction of COUP-TFII into HeLa cells also decreased both endogenous hTERT transcription and telomerase activity. Conclusion The human COUP-TFII can firmly bind to hTERT promoter, and inhibit telomerase activity through decreasing hTERT transcription. It will greatly facilitate understanding of telomerase regulation in normal and cancer cells
文摘Objective: To investigate the effects of lithium on cognitive function and metabolism of Amyloid-beta Protein Precursor (APP) and tau phosphorylation in rats chronically exposed to aluminum. Methods: Twenty-four chronically aluminum-exposed rats were randomly divided into 2 groups: a lithium-treatment group and a non-treatment group (n=12 per group). Lithium chloride was administered to the lithium-treatment group via gastric gavage daily for 6 weeks (200 mg/kg·d LiCl), while the non-treatment group was administered the same volume of sodium chloride by the same means. An additional control group (n=12) received no intervention. Memory function was evaluated by the Morris water maze test. Aβ was measured by immunohistochemical staining, while total APP, phosphorylated-tau protein, CDK5 and PP2A were determined by Western Blotting. Results: (1) Compared to the non-treatment group, the lithium-treatment group had a significantly shorter mean escape latency and a lower proportion of random navigation pattern in the spatial probe test (P<0.05). After the platform was taken away, the rats in the lithium-treatment group crossed the platform quadrant significantly more and stayed longer in the platform quadrant than those in the non-treatment group (P<0.05). (2) The number of Aβ positive neurons in the hippocampus and cortex was significantly less in the lithium-treatment group than in the non-treatment group (P<0.05), but the content of APP was not different between groups (P=0.730). (3) Phosphorylation of tau protein decreased significantly in the lithium-treatment group than that in the non-treatment group (P<0.05). The content of CDK5 in the lithium-treatment group was significantly less than that in the non-treatment group in the cortex and hippocampus, while there was no difference in the content of PP2A between the 2 groups. The expression of CDK5 was significantly correlated with phosphorylated tau (r=0.871, P=0.024) in the lithium-treatment group. Conclusion: Lithium may improve memory function in rats chronically exposed to aluminum by decreasing both the production of Aβ and tau phosphorylation, with the latter results from inhibiting expression of CDK5.
基金Supported by grants from the Hunan Natural Science foundation (No.06JJ4199)the Hunan Science Technology Foundation (No.2007SK3010)
文摘Objective: To investigate the proteornic differences between the high-sensitivity (HS) group and low-sensitivity (LS) group of cervical cancer treated by radiotherapy and confirm the radiotherapy sensitivity associated proteins in early cervical cancer. Methods: The fresh carcinoma tissues were collected from 10 untreated cervical cancer patients and preserved in the -80 ℃ refrigeratory. The tissues were classified into two groups: high sensitivity group (HS) and low sensitivity group (LS), according to their response to radiotherapy. In the first part of our experiment, protein separating was performed by using two-dimensional gel electrophoresis (2-DE) with Amersham 18 cm linear pH 3-10 immobilized pH gradient (IPG) strips. The images of the gels were acquired by the scanner and then analyzed by using PD-quest7.3 software to find the differentially expression protein-spots in each group. Then the differentially expressed protein-spots was incised from the gels and digested by trypsin. The peptide mass fingerprintings (PMF) was acquired by matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and the proteins were identified by data searching in the Mascot-database. Part of differentially expression proteins were assayed by Western Blot. Results: Most of the gels were clear and successfully analyzed by PD-quest7.3 software. Most of the protein-spots concentrated on the area of 20-100Kda (Mw) and pH4-8. The average number of the protein-spots was 754 ± 64 in HS group and 777 ±48个 in LS group. The match rate was 87.6% between two groups. Five high expression proteins were found in HS group which were low expression in LS group, 3 high expression protein were found in LS group which were low expression in HS group. Reselts of Western Blot were in coincidence to proteomic result. Conclusion: The 2-DE gels image of HS group and LS group with early cervical cancer tissues treated by radiotherapy are successfully acquired. Some differentially expression proteins between the two groups are further confirmed by immunohistochemical assay.
基金Supported by a grant from the Medical and Health Technology Development Program in Shandong Province(No.2015WS0407)
文摘Objective This study aimed to combine tumor abnormal protein(TAP) and high-sensitivity C-reactive protein(hs-CRP) level detection to diagnose endometrial cancer in patients with endometrial thickness less than 8 mm, and to provide a reference for clinical screening and diagnosis. Methods Clinical data from 19 cases of endometrial cancer, diagnosed on the basis of pathological findings, were collected from September 2014 to December 2015. The inclusion criteria were as follows: the patients were first diagnosed with endometrial thickness less than 8 mm and were all in menopause. Perimenopausal patients(n = 26) with uterine fibroids seen during the same period were selected as a control group. Serum TAP and hs-CRP levels of the patients in the two groups were simultaneously determined on admission. Results We found that both TAP and hs-CRP levels in the experimental group were higher than those in the control group [(182.95 ± 72.14) μm^2 vs.(133.19 ± 55.18) μm^2, P = 0.019;(7.52 ± 19.03) mg/L vs.(1.66 ± 2.31) mg/L, P = 0.136]. The sensitivity of TAP for the diagnosis of endometrial cancer was 73.68%, the specificity was 69.23%, and the Youden index was 0.4291. The diagnostic sensitivity and specificity of hs-CRP was 15.79% and 100%, respectively, and the Youden index was 0.1579. After plotting the receiver operating characteristics curves, the optimal cut-off value for TAP in diagnosing endometrial cancer was found to be 160.662 μm^2 and that for hs-CRP was 1.07 mg/L. Conclusion For patients suspected of having endometrial cancer with endometrial thickness less than 8 mm, combined detection of TAP and hs-CRP levels can be used as a screening tool and can provide new ideas regarding clinical diagnosis and treatment.
基金Supported by Research Project for Traditional Chinese Medicine of Shaanxi Administration of Traditional Chinese Medicine (jc87)Program for Changjiang Scholars and Innovative Research Team in University (PCSIRT: 1171)
文摘Objective: To retrospectively analyze the clinical data of 275 cases of cervical cancer (CC) in our hospital, and investigate the clinicopathological parameters of cervical cancer based on the expression of p16INK4A proteins. Methods: The clinical information of 275 patients with cervical cancer were retrospectively analyzed between 2006 and 2011, including the patients' age, clinical FIGO stage, differentiation, histologic grade, infiltration depth, treatment, pathological diagnosis after surgery, and results of following-up.Immunohistochemistry was also done on sections of confirmed cancer specimens without prior chemotherapy/radiotherapy. Results: Among various clinicopathological parameters, the median age was 48 years old. The grade was significantly associated with histological type, HPV infection and with lymph node invasion. FIGO stage was strongly correlated to the infiltration depth and lymph node metastasis. P16INK4A expression was significantly correlated with histologic grade. However, there were no differences between p16INK4A staining and patient's age, histopathology and lymph node metastases. Conclusion: The incidence of cervical cancer becomes increasingly younger. Additionally, p16INK4A can function as a diagnostic marker of cervical carcinomas.
文摘To investigate the effects of early nutritional intervention on the serum insulin-like growth factor-1 (IGF1),insulin-like growth factor binding protein 3 (IGFBP3), intestinal development, and catch-up growth of intrauterine growth retardation (IUGR) rats by giving the IUGR new born rats different protein level diet. Methods IUGR rat model was built by starvation of pregnant female rats. Twenty-four IUGR pups and 8 normal pups were divided randomly into 4 groups: normal control group (C group); IUGR control group(S group), IUGR low-protein diet group (SL group), and IUGR high-protein diet group (SH group). Detected the serum IGF1, IGFBP3, body weight, body length, intestinal weight length, intestinal villi height (VH), crypt depth (CD), villi absorbing area (VSA), mucous thickness (MT), and disaccharidase at the 4th week. Results (1) The SH group showed the fastest catch-up growth, serum IGF1, IGFBP3, VH, and VSA were significantly higher than those of normal control group and IUGR control group. The intestinal weight and length, and the activities of lactase and saccharase of the SH group also reached the normal control group level. (2) The SL group kept on small size, the serum IGF1, IGFBP3, and most of intestinal histological indexes were all significantly lower than other groups. (3) IGF-1, IGFBP3 were positively correlated to intestinal VH, VSA, saccharase, body weight and length. Conclusions The serum IGF1 was a sensitive index to the catch-up growth. The early nutritional intervention of high-protein diet after birth is helpful for the catch-up growth of IUGR through promoting the intestinal development and the ab-sorption of nutrition
基金Project supported by the National Natural Science Foundation of China(No.31172206)the Grant-in-Aid for Innovative Training of Doctoral Students in Jiangsu Province(No.CXLX13-287)
文摘The objective was to investigate the expression of bone morphogenetic protein (BMP) family members in the mouse uterus during the estrous cycle by real-time polymerase chain reaction (PCR) and immunohistochemistry. Uterine samples from Swiss ICR mice were collected and dissected free of surrounding tissue. One uterine horn was snap frozen in liquid nitrogen immediately after collection and stored at -80 ℃for RNA extraction, and the other was fixed in 40 mg/ml paraformaldehyde at room temperature for immunolocalization of BMP2 protein. Real-time PCR analysis showed that the expression level of Bmp2 was significantly higher at proestrus than at estrus and metestrus (P〈0.05). The relative abundance of Bmp4 exhibited significant fluctuations, but there were no statistically significant differences between the expression levels of Bmp2 and Bmp4 (P〉0.05). The expression levels of Bmprla and Bmpr2 remained unchanged during estrous cycles. However, the level of Bmprlb mRNA decreased significantly at estrus (P〈0.05), increasing subsequently at metestrus. Furthermore, the level of Bmprlb mRNA was significantly lower than those of Bmprla and Bmpr2 mRNA at the corresponding stages (P〈0.05). All three receptor-regulated Smads (R-Smads) detected were differentially expressed in the mouse uterus and the expression levels of Smadl and Smad5 were significantly higher than that of Smad8 (P〈0.05). In addition, the expression level of Smad4 did not change substantially throughout the estrous cycle. Immunohistochemical experiments revealed that BMP2 protein was differentially expressed and localized mainly in the uterine luminal and glandular epithelial cells throughout the estrous cycle. In conclusion, our results provide information about the variation in the mRNA levels of Bmp2 and Bmp4 and related components of the BMP signaling pathway. The data provide quantitative and useful information about the roles of endometrial BMP proposed and demonstrated by others, such as the degradation and remodeling of the endometrium.
基金Supported by the National Science and Technology "12th Five-years" Major Special-purpose Foundation (No 2011ZX09201-201-01)
文摘OBJECTIVE:To observe the effects of Compound Danshen Tablets(CDST) on spatial cognition and expression of brain b-amyloid precursor protein(β-APP) in a rat model of Alzheimer's disease.METHODS:The rat model of Alzheimer's disease(AD) was established using D-galactose to cause subacute aging combined with Meynert nucleus damage.Rat behavior was monitored using the Morris water maze,and the expression of β-APP in rat brain tissue was detected via immunohistochemistry.RESULTS:CDST significantly improved spatial cognition and decreased β-APP expression in the cortex and hippocampus(P<0.05,P<0.01).CONCLUSIONS:CDST can significantly improve spatial cognition in a rat model of AD.This observation is possibly related to a reduction in β-APP ex-pression in the rat brain.
基金supported by the National Natural Science Foundation of China(No.30700213)National Basic Research Development Program of China(No.2007CB947804)
文摘Objective It has been shown that there are extensive interactions between the central nervous system and the immune system.The present study focused on the effects of lipopolysaccharide(LPS)on memory retrieval,to explore the interaction between immune activation and memory.Methods C57BL/6J mice(8 weeks old)were first trained in the Morris water maze to reach asymptotic performance.Then mice were tested 24 h after the last training session and LPS was administered(1.25 mg/kg,i.p.)4 h prior to the testing.The retrieval of spatial memory was tested by probe trial,and the time spent in the target quadrant and the number of platform location crosses were recorded.ELISA was performed to detect interleukin-1β(IL-1β)protein level in the hippocampus of mice tested in the water maze.Results Although LPS induced overt sickness behavior and a significant increase in the level of IL-1β in the hippocampus of mice,there was no significant difference in the time spent in the target quadrant or in the number of platform location crosses between LPS-treated and control groups in the probe trial testing.Conclusion Immune activation induced by LPS does not impair the retrieval of spatial memory.
基金This work was supported by the National Institute of Health(NIH R01-HD067449)。
文摘Maternal stress during pregnancy is prevailing worldwide, which exposes fetuses to intrauterine hyper glucocorticoids(GC), programming offspring to obesity and metabolic diseases. Despite the importance of brown adipose tissue(BAT) in maintaining long-term metabolic health, impacts of prenatal hyper GC on postnatal BAT thermogenesis and underlying regulations remain poorly defined. Pregnant mice were administrated with synthetic GC dexamethasone(DEX) at levels comparable to fetal GC exposure of stressed mothers. Prenatal GC exposure dose-dependently reduced BAT thermogenic activity, contributing to lower body temperature and higher mortality of neonates;such difference was abolished under thermoneutrality, underscoring BAT deficiency was the major contributor to adverse changes in postnatal thermogenesis due to excessive GC. Prenatal GC exposure highly activated Redd1 expression and reduced Ppargc1 a transcription from the alternative promoter(Ppargc1 a-AP) in neonatal BAT. During brown adipocyte differentiation, ectopic Redd1 expression reduced Ppargc1 a-AP expression and mitochondrial biogenesis;and the inhibitory effects of GC on mitochondrial biogenesis and Ppargc1 a-AP expression were blocked by Redd1 ablation. Redd1 reduced protein kinase A phosphorylation and suppressed cyclic adenosine monophosphate(c AMP)-responsive element-binding protein(CREB) binding to the c AMP regulatory element(CRE) in Ppargc1 a-AP promoter, leading to Ppargc1 a-AP inactivation. In summary, excessive maternal GC exposure during pregnancy dysregulates Redd1-Ppargc1 a-AP axis, which impairs fetal BAT development, hampering postnatal thermogenic adaptation and metabolic health of offspring.
基金Supported by the Innovation Fundation of Chinese Medicine research,Guangzhou University of Chinese Medicine!Effect of Chinese Medicines in the Warming Yang and Removing Blood Stasis Method on the Biological Characteristics of Cultured Endometrial Cells from Patients with Endometriosis,No.K0080053)the Natural Science Foundation of Heibei Province of China(Effect of Dang Gui Si Ni Jia Wu Zhu Yu Sheng Jiang Huang Jiu Decoction via NF-KBp65signaling pathway on the regulating endometrial Biological Characteristics of endometriosis.No.2015CFB590)
文摘OBJECTIVE:To investigate the effect of Chinese medicines using the warming Yang and removing blood stasis method on levels of matrix metalloproteinases(MMPs)/tissue inhibitor metalloproteinases(TIMPs) secreted by cultured endometrial cells from patients with endometriosis.METHODS:Ectopic and eutopic endometrial cells obtaind from 15 endometriosis patients were cultured in vitro,and divided randomly into five groups:high dose;moderate dose;low dose;nemestran;blank control.The three dose groups were treated with a decoction prepared according to the principle of warming Yang and removing blood stasis;nemestran and 0.9%NaCI were administered to the nemestran group and balnk control group,respectively.Eutopic endometrial cells obtaind from 10 hysteromyoma patients were cultured in vitro,as the normal control group,0.9%NaCI were administered to the normal control group.Cell culture supernatants were collected and levels of matrix metalloproteinase-1(MMP-1),matrix metalloproteinase-2(MMP-2),matrix metalloproteinase-9(MMP-9),tissue inhibitor metalloproteinase-1(TIMP-1) and tissue inhibitor metalloproteinase-2(TIMP-2) detected by enzyme-linked immuno sorbent assay(ELISA).RESULTS:Compared with the normal control group,levels of MMP-1,MMP-2,and MMP-9 in eutopic and ectopic endometrium cell supernatants in the blank control group were increased,whereas levels of TIMP-1 and TIMP-2 were decreased(P <0.05).Compared with the blank control group,levels of MMP-1 and MMP-2 in ectopic and eutopic endometrium cell supernatants cultured in low-dose,middle-dose,and high-dose groups were decreased,whereas levels of TIMP-1 and TIMP-2 were increased significantly(P < 0.05).CONCLUSION:The warming Yang and removing blood stasis method affects expression of MMPs andTIMPs.