达格列净对Ⅱ型心肾综合征患者心肾功能及短期预后的影响

目的 观察在常规治疗基础上,加用达格列净对Ⅱ型心肾综合征(cardiorenal syndrome, CRS)患者的血清肌酐(serum creatinine, SCr)、血胱抑素C(cystatin C, Cys-C)、可溶性生长激素表达基因2蛋白(growth stimulation expressed gene 2, sST2)和N末端B型利钠肽原(n-terminal pro b-type natriuretic peptide, NT-proBNP)的影响。方法 选取Ⅱ型CRS患者94例,随机分为对照组48例(常规治疗)和观察组46例(常规治疗的基础上加用达格列净10 mg),在基线与治疗24周后检测心肾功能指标,并观察短期预后以及不良反应。结果 与对照组比较,观察组患者的SCr、Cys-C、sST2和NT-proBNP值均明显降低(P<0.05),观察组主要不良心血管事件发生率更低(P<0.05)。结论 在常规治疗基础上加用达格列净10 mg可显著降低Ⅱ型CRS患者的SCr、Cys-C、sST2和NT-proBNP水平及改善短期预后,且sST2可用来判断Ⅱ型CRS患者心肾功能,有助于指导诊疗。

Study on the Developmental Changes of Muscular GHR mRNA Expression in Sheep

[ Objective] The study aimed to explore the expression of muscular growth hormone receptor gene （GHR） in sheep at the early stage of growth and development. [Method] The GHR mRNA expression levels in longissimus dorsal muscles of male Kazak sheep and Xinjiang fine wool sheep with different ages were quantitatively analyzed by real time PCR. [ Result] Sheep GHR mRNA expression level in longissimus dorsal muscle increased firstly followed by decline, and then kept steady until the end of the experiment, with the expression peak appearing on postnatal day 30. The GHR mRNA expression level of Kazak sheep was extremely lower than that of Xingjiang fine wool sheep from 2 to 90 days old （ P 〈0.01 ）. E Conclusionl Both age and breed had great effects on the expression of muscular GHR gene in sheep.

生理妊娠(正常妊娠)

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Correlation between expression of gastrin, somatostatin and cell apoptosis regulation gene bcl-2/bax in large intestine carcinoma

AIM: To explore the correlation between expression of somatostatin (SS), gastrin (GAS) and cell apoptosis regulation gene bcl-2/bax in large intestine carcinoma.METHODS: Sixty-two large intestine cancer tissue samples were randomly and retrospectively selected from patients with large intestine carcinoma. Immunohistochemical staining for bcl-2, bax, GAS, SS was performed according to the standard streptavidin-biotin-peroxidase (S-P) method.According to the semi-quantitative integral evaluation, SS and GAS were divided into three groups as follows. Scores1-3 were defined as the low expression group, 4-8 as the intermediate expression group, 9-16 as the high expression group. Bax and bcl-2 protein expressions in different GAS and SS expression groups of large intestine carcinoma were assessed.RESULTS: The positive expression rate of bax had a prominent difference between SS and GAS high, intermediate and low expression groups (P＜0.05, x2ss = 9.246; P＜0.05,x2GAS = 6.981). The positive expression rate of bax in SS high (80.0%, 8/10) and intermediate (76.5%, 13/17)expression groups was higher than that in low expression group (40.0%, 14/35) (P＜0.05, x2high vs low = 5.242; P＜0.05,x2middle vs low = 6.097). The positive expression rate of bax in GAS high expression group (27.3%, 3/8) was lower than that in low expression group (69.4%, 25/36) (P＜0.05,x2 = 4.594). However, bax expression in GAS intermediate expression group (46.7%, 7/15) was lower than that in low expression group, but not statistically significant. The positive expression rate of bcl-2 had a prominent difference between SS and GAS high, intermediate and low expression groups (P＜0.05, x2ss = 7.178; P＜0.05, x2GAS = 13.831). The positive expression rate of bcl-2 in GAS high (90.9%, 10/11)and intermediate (86.7%, 13/15) expression groups was higher than that in low expression group (44.4%, 16/36)(P＜0.05,x2high vs low = 5.600; P＜0.05, x2 middle vs low = 7.695).However, the positive expression rate of bcl-2 in SS high (40.0%, 4/10) and intermediate (47.1%, 8/9) expression groups was lower than that in low expression group (77.1%, 27/35)(P＜0.05, x2 high vs low = 4.710; P＜0.05, x2 middle vs low = 4.706).There was a significant positive correlation between the integral ratio of GAS to SS and the integral of bcl-2 (P＜0.01,r=0.340). However, there was a negative correlation between the integral ratio of GAS to the SS and bax the integral of (P＜0.05, r = -0.299).CONCLUSION: The regulation and control of gastrin,somatostatin in cell apoptosis of large intestine carcinoma may be directly related to the abnormal expression of bcl-2, bax.

Effect of electro-acupuncture at Foot-Yangming Meridian on somatostatin and expression of somatostatin receptor genes in rabbits with gastric ulcer

AIM： To discuss the protective effect of electroacupuncture at the Foot-Yangming Meridian on gastric mucosal lesion, somatostatin （SS） and the expression of SS receptor genes （SSR1mRNA） in rabbits with gastric ulcer and to further explore the relative specificity of meridians and viscera at gene expression level. METHODS： Forty rabbits were randomly divided into control group （A）, gastric ulcer model group （B）, FootYangming Meridian group （C）, Foot-Shaoyang Meridian group （D） and Foot-Taiyang Meridian group （E）. The gastric ulcer model was prepared by infusing alcohol into stomach. Groups C-E were treated with electroacupuncture at points along the above meridians using meridian stimulating instruments for 7 days respectively. By the end of treatment, the index of gastric ulcer was determined, the amount of epidermal growth factor（EGF） and somatostatin was measured by radioimmunoassay （PJA）. SS-R1mRNA expression in gastric mucosa was determined by RT-PCR. RESULTS： The value of EGF in model group was obviously lower（73.6±14.8 vs 91.3±14.9 pg/mL, P〈 0.01） than that in control group. The index of gastric ulcer, content of SS and expression of SSRtmRNA in gastric mucosa were significantly higher than those in control group （24.88 ± 6.29 vs 8.50 ± 2.98 scores, P〈 0.01; 2978.6 ± 587.6 vs 1852.4 ± 361.7 mIU/mL, P〈 0.01; 2.56±0.25 vs 1.04±0.36, P〈0.01） . The value of EGF in Foot-Yangming Meridian group was higher than that in model group（92.2±6.7 vs 73.6±14.8 pg/mL, P〈 0.01）. The index of gastric ulcer, content of SS and expression of SS-R1mRNA in gastric mucosa were significantlylower than those in control group（10.88±3.23 vs 24.88±6.29 scores, P〈0.01; 1800.2±488 vs 2978.6±587.6 mIU/mL, P 〈 0.01; 1.07±0.08 vs 2.56±0.25mIU/mL, P〈0.01）. Compared to the model group, the content of SS and expression of SSRlmRNA in gastric mucosa in Foot-Shaoyang Meridian group decreased （2441.0±488. vs 2978.6± 587.6 mIU/mL, P〈 0.05; 1.73±0.16 vs 2.56±0.25 mIU/mL, P〈0.01）. But the above parameters in Foot-Taiyang Meridian group did not improve and were significantly different from those in Footoyangming Meridian group （P〈0.05） CONCLUSION： Electro-acupuncture at Foot-Yangming Meridian can protect gastric mocusa against injury. The mechanism may be releted to the regulation of brain-gut peptides and the expression of SSRtmRNA.

Correlation between the expressions of gastrin, somatostatin and cyclin and cyclin-depend kinase in colorectal cancer

AIM： To explore the correlation between the expressions of gastrin （GAS）, somatostatin （SS） and cyclin, cyclin- dependent kinase （CDK） in colorectal cancer, and to detect the specific regulatory sites where gastrointestinal hormone regulates cell proliferati6n. METHODS： Seventy-nine resected large intestine carcinomatous specimens were randomly selected. Immunohistochemical staining for GAS, SS, cyclin D1, cyclin E, cyclin A, cyclin B1, CDK2 and CDK4 was performed according to the standard streptavidinbiotin-peroxidase （S-P） method. According to the semiquantitative integral evaluation, SS and GAS were divided into high, middle and low groups. Cyclin D1, cyclin E, cyclin A, cydin B1, CDK2, CDK4 expressions in the three GAS and SS groups were assessed. RESULTS： The positive expression rate of cyclin D1 was significantly higher in high （78.6%, 11/14） and middle GAS groups （73.9%, 17/23） than in low GAS group （45.2%, 19/42） （P〈0.05, X^2 high vs low = 4.691; P〈0.05, X^2 middle vs low = 4.945）. The positive expression rate of cyclin A was significantly higher in high （100%, 14/14） and middle GAS groups （82.6%, 19/23） than in low GAS group （54.8%, 23/42） （P〈0.01, X2high vs low = 9.586; P〈0.05, X^2 middle vs low = 5.040）. The positive expression rate of CDK2 was significantly higher in high （92.9%, 13/14） and middle GAS groups （87.0%, 20/23） than in low GAS group （50.0%, 21142） （P〈0.01, X^2 high vs low = 8.086; P〈0.01,X^2 middle low = 8.715）. The positive expression rate of CDK4 was significantly higher in high （78.6%, 11/14）and middle GAS groups （78.3%, 18/23） than in low GAS group （42.9%, 18/42） （P〈0.05, X^2 high vs low= 5.364; P〈0.01, X^2 middle vs low = 7.539）. The positive expression rate of cyclin E was prominently higher in low SS group （53.3%, 24/45） than in high （9.1%, 1/11） and middle （21.7%, 5/23） SS groups （P〈0.05, X^2 high vs low = 5.325; P〈0.05, X^2 middle vs low = 6.212）. The positive expression rate of CDK2 was significantly higher in low SS group （77.8%, 35/45） than in high SS group （27.3%, 3/11） （P〈0.01, X^2 high vs low = 8.151）. There was a significant positive correlation between the integral ratio of GAS to SS and the semi-quantitative integral of cyclin D1, cyclin E, cyclin A, CDK2, CDK4 （P〈0.05, 0% = 0.252; P〈0.01, E^rs = 0.387; P〈0.01,A^rs = 0.466; P〈0.01, K2^rs = 0.519; P〈0.01, K4^rs = 0.434）. CONCLUSION： The regulation and control of gastrin, SS in colorectal cancer cell growth may be directly related to the abnormal expressions of cyclins D1, A, E, and CDK2, CDK4. The regulatory site of GAS in the cell cycle of colorectal carcinoma may be at the G2, S and G2 phases. The regulatory site of SS may be at the entrance of S phase.

Effects of intra-gastric beta-casomorphin-7 on somatostatin and gastrin gene expression in rat gastric mucosa

AIM： To investigate the in vivo effect of beta-casomorphin-7on the regulation of gastric somatostatin and gastrin messenger RNA in rat gastric mucosa.METHODS： Somatostatin and gastrin mRNA were quantified by RT-PCR and in situ hybridization （ISH）in 24 rats. The rats were divided into three treatment groups： basal diet ＋ physiological saline （n = 8）, basal diet ＋ beta-casomorphin-7 （7.5 × 10^-7 mol） （n = 8）,and basal diet ＋ poly-Gly-7 （containing equal mol of N with 7.5 × 10^-7 mol beta-casomorphin-7） （n = 8）.After oral administration for 30 days, rats were killed by exsanguinations.RESULTS： After intra-gastric administration of betacasomorphin-7 for 30 d, gastrin mRNA increased by 52.8% （P 〈 0.05, n = 8）, and somatostatin mRNA levels decreased by 30.7% compared with the controls （P 〈0.01, n = 8）. No significant differences in the expression of the two genes were observed in the poly-Gly-treated group, although gastrin mRNA expression was elevated by 35.6% as against the control group （P = 0.15, n =8）. The long-term oral administration of a casomorphin solution significantly decreased the even gray of D-cells,but did not lower the number of D-cells both in the antrum and fundus. Interestingly, the number of G-cells increased in the antrum and fundus, but its average density was augmented only in the antrum.CONCLUSION： Beta-casomorphin-7 is capable of modulating gene expression of the regulatory peptides from G and D cells. Data from in situ hybridization studies indicate that beta-casomorphin-7 affects gastrin gene expression indirectly by means of the paracrine action of somatostatin, and depends on its intrinsic molecular function.

Expression of Porcine Growth Hormone Gene in CHO Cell

An expressive plasmid pSMTPCH was constructed from porcine growth hormone gene,sheep metallothionein promoter (MT-011)and the vector,pUC19. The linear pSMTPGH and circular pSV2-dhfr were cotransfected into CHO-dhfr cell by calcium phosphate coprecipitation. Positive clones made up 74% of total clones, which were identified with ELISA. The expression of pSMTPGH was induced by 0.5 μM of Cd￣++. The clone 1-C-3 was found to secrete hGH at the level of 3800 μg/10￣6 cells/24 hrs in media containing 10 μMTX. After 20 generations in culture, the clone was still stable with hGH expression.The molecular weight of secreted protein was the same as that of the natural pGH, 22KD;the identity was further supported by Western blot.