Expression of lung resistance protein in patients with gastric carcinoma and its clinical significance

INTRODUCTION The efficacy of chemotherapy in the treatment of cance patients is often hampered by the presence or appearance of multidrug resistance(MDR) of tumor cells.

Positive Selection Analysis of VP1 Genes of Worldwide Human Enterovirus 71 Viruses

Human enterovirus 71 viruses have been long circulating throughout the world. In this study, we performed a positive selection analysis of the VP1 genes of capsid proteins from Enterovirus 71 viruses. Our results showed that although most sites were under negative or neutral evolution, four positions of the VP1 genes were under positive selection pressure. This might account for the spread and frequent outbreaks of the viruses and the enhanced neurovirulence. In particular, position 98 might be involved in neutralizing antibodies, modulating the virus-receptor interaction and enhancing the virulence of the viruses. Moreover, both positions 145 and 241 might correlate to determine the receptor specificity. However, these positions did not display much difference in amino acid polymorphism. In addition, no position in the VP1 genes of viruses isolated from China was under positive selection.

Gastric mucosal damage in water immersion stress:Mechanism and prevention with GHRP-6

AIM:To investigate the mechanism of gastric mucosal demage induced by water immersion restraint stress(WRS) and its prevention by growth hormone releasing peptide-6(GHRP-6).METHODS:Male Wistar rats were subjected to conscious or unconscious(anesthetized) WRS,simple restraint(SR),free swimming(FS),non-water fluid immersion,immersion without water contact,or rats were placed in a cage surrounded by sand.To explore the sensitivity structures that influence the stress reaction besides skin stimuli,a group the rats had their eyes occluded.Cervical bilateral trunk vagotomy or atropine injection was performed in some rats to assess the parasympathetic role in mucosal damage.Gastric mucosal lesions,acid output and heart rate variability were measured.Plasma renin,endothelin-1 and thromboxane B2 and gastric heat shock protein 70 were also assayed.GHRP-6 was injected [intraperitoneal(IP) or intracerebroventricular(ICV)] 2 h before the onset of stress to observe its potential prevention of the mucosal lesion.RESULTS:WRS for 6 h induced serious gastric mucosal lesion [lesion area,WRS 81.8 ± 6.4 mm 2 vs normal control 0.0 ± 0.0 mm 2,P < 0.01],decreased the heart rate,and increased the heart rate variability and gastric acid secretion,suggesting an increase in vagal nervecarrying stimuli.The mucosal injury was inversely correlated with water temperature(lesion area,WRS at 35 ℃ 56.4 ± 5.2 mm 2 vs WRS at 23 ℃ 81.8 ± 6.4 mm 2,P < 0.01) and was consciousness-dependent.The injury could not be prevented by eye occlusion,but could be prevented by avoiding contact of the rat body with the water by dressing it in an impermeable plastic suit.When water was replaced by vegetable oil or liquid paraffin,there were gastric lesions in the same grade of water immersion.When rat were placed in a cage surrounded by sand,there were no gastric lesions.All these data point to a remarkable importance of cutenuous information transmitted to the high neural center that by vagal nerves reaching the gastric mucosa.FS alone also induced serious gastric injury,but SR could not induce gastric injury.Bilateral vagotomy or atropine prevented the WRS-induced mucosal lesion,indicating that increased outflow from the vagal center is a decisive factor in WRS-induced gastric injury.The mucosal lesions were prevented by prior injection of GHRP-6 via IP did,but not via ICV,suggesting that the protection is peripheral,although a sudden injection is not equivalent to a physiological release and uptake,which eventually may affect the vagal center.CONCLUSION:From the central nervous system,vagal nerves carry the cutaneous stimuli brought about by the immersion restraint,an experimental model for inducing acute gastric erosions.GHRP-6 prevents the occurrence of these lesions.

Seven-senescence-associated gene signature predicts overall survival for Asian patients with hepatocellular carcinoma

BACKGROUND Cellular senescence is a recognized barrier for progression of chronic liver diseases to hepatocellular carcinoma(HCC). The expression of a cluster of genes is altered in response to environmental factors during senescence. However, it is questionable whether these genes could serve as biomarkers for HCC patients.AIM To develop a signature of senescence-associated genes(SAGs) that predicts patients' overall survival(OS) to improve prognosis prediction of HCC.METHODS SAGs were identified using two senescent cell models. Univariate COX regression analysis was performed to screen the candidate genes significantly associated with OS of HCC in a discovery cohort(GSE14520) for the least absolute shrinkage and selection operator modelling. Prognostic value of this seven-gene signature was evaluated using two independent cohorts retrieved from the GEO(GSE14520) and the Cancer Genome Atlas datasets, respectively.Time-dependent receiver operating characteristic(ROC) curve analysis was conducted to compare the predictive accuracy of the seven-SAG signature and serum α-fetoprotein(AFP).RESULTS A total of 42 SAGs were screened and seven of them, including KIF18 B, CEP55,CIT, MCM7, CDC45, EZH2, and MCM5, were used to construct a prognostic formula. All seven genes were significantly downregulated in senescent cells andupregulated in HCC tissues. Survival analysis indicated that our seven-SAG signature was strongly associated with OS, especially in Asian populations, both in discovery and validation cohorts. Moreover, time-dependent ROC curve analysis suggested the seven-gene signature had a better predictive accuracy than serum AFP in predicting HCC patients' 1-, 3-, and 5-year OS.CONCLUSION We developed a seven-SAG signature, which could predict OS of Asian HCC patients. This risk model provides new clinical evidence for the accurate diagnosis and targeted treatment of HCC.

Effects of polygonatum sibiricum polysaccharide on learning and memory in a scopolamine-induced mouse model of dementia

BACKGROUND： Learning and memory processes are accompanied by complex neuropathological and biochemical changes. Free radicals play an important role in learning and memory damage. OBJECTIVE： To observe the effects of polygonatum sibiricum polysaccharide （PSP） in comparison with vitamin 12 on inhibiting free radical damage, as well as improving the degree of cerebral ischemia and learning and memory in a scopolamine-induced mouse model of dementia. DESIGN： Randomized controlled animal study. SETTINGS： Department of Pharmacology, Taishan Medical College; Shandong Jewim Pharmaceutical Co., Ltd. MATERIALS： A total of 105 healthy Kunming mice, comprising 90 males and 15 females that were clean grade, were provided by the Animal Center of Taishan Medical College. PSP （extracted and purified by Huangjing, Taishan） was provided by the Department of Traditional Chinese Medicine, Taishan Medical College （purity of 79.6% by using a phenol-concentrated sulphate acid method）, and hydrogen bromine acid scopolamine injection solution （SCO） by Shanghai Hefeng Pharmaceutical Co., Ltd. METHODS： This study was performed at the Pharmacological Laboratory of Taishan Medical College from March to June 2007. （1） A total of 75 healthy Kunming male mice of clean grade were randomly divided into a normal control group, positive control group, and low-dosage and high-dosage PSP groups, with 15 mice in each group. Mice in both the low-dosage and high-dosage PSP groups were intragastrically administered 0.5 g/kg and 2.0 g/kg PSP, respectively. Mice in the positive control group were intragastrically administered 0.5 g/kg vitamin 12. In addition, mice in both the normal control group and model group were intragastrically administered the same volume of saline, respectively, once a day for 7 consecutive days. One hour after the final administration on day 6, mice in the positive control group, model group, low-dosage and high-dosage PSP groups were subcutaneously injected with 3.0 mg/kg SCO, while mice in the normal control group were subcutaneously injected with the same volume of distilled water. Ten minutes later, the step test was employed to measure memory. The training was performed 5 times, with 30-minute intervals between 2 sets. If the mice remained on the platform （latent period） for 30 minutes, they were determined to have learned the task. An eligible percentage was then recorded. Twenty-four hours later, the number of error responses from each mouse was recorded in a 5-minute period, based on the above-mentioned parameters. Mice were sacrificed under anesthesia. The activities of glutathione hyperoxide enzyme （GSH-Px）, superoxide dismutase （SOD）, and the content of malondialdehyde （MDA） were assayed using an UV spectrophotometer. （2） The remaining 30 healthy Kunming mice of both genders were randomly divided into 3 groups, including control group, low-dosage PSP group, and high-dosage PSP group, with 10 mice in each group. Mice in both the low-dosage and high-dosage PSP groups were intragastrically administered 0.5 g/kg and 2.0 g/kg PSP, respectively, while the mice in the control group were perfused with the same volume of saline. Forty minutes later, the mice under superficial anesthesia were decapitated, and the number and duration of mouth-opening breaths of the isolated mouse head were immediately recorded. MAIN OUTCOME MEASURE： （1） Numbers of error responses within 5 minutes on the platform. （2） GSH-Px and SOD activity, as well as MDA content in mouse brain tissue. （3） Numbers and duration of mouth-opening breaths of the isolated mouse head. RESULTS： Of the 105 Kunming experimental mice, two mice died due to electric shock during the step-down test, therefore, a total of 103 mice were involved in the final analysis. （1） Effects of PSP on learning in mice： The eligible percentage in the high-dosage PSP group was higher than the control group at the 3rd and 5th training sessions （P 〈 0.05）. （2） Effects of PSP on memory in mice： The number of errors in the step-down test in the model group was higher than in the normal control group （P 〈 0.01）. Compared to the model group, the number of errors in the step-down test was lower in both the low-dosage and high-dosage PSP groups （P 〈 0.01）. （3） Effects of PSP on amount of GSH-Px, SOD, and MDA in mouse brain tissue： SOD and GSH-Px activity was higher in both the low-dosage and high-dosage PSP groups than in the model group. MDA content was lower in the high-dosage PSP group, compared to the model group. GSH-Px activity in the brain tissue of the high-dosage PSP group was similar to the positive control group （P 〉 0.05）. （4） Effects of PSP on acute cerebral ischemia in mice： The low-dosage PSP, and in particular the high-dosage PSP, prolonged the number and duration of mouth-opening breaths of the isolated mouse head （P 〈 0.05, 0.01）. CONCLUSION： PSP can improve learning and memory in a scopolamine-induced mouse model of dementia by reducing the damaging effects of cerebral ischemia and anti-oxidation. In addition, the effects are dose-dependent and are similar to those provided by vitamin E.

PCR Detection and Sequence Analysis of Duck Circovirus in Sick Muscovy Ducks

在从福建省的病了的鸭的鸭 circovirus (DuCV ) 感染被调查。有传染 serositis 的 43 只病了的俄国鸭的肝样品在福建省从 12 个鸭农场被收集。基于 DuCV 的出版序列，二份教材为 DuCV 的察觉被设计，四份教材被设计放大盖住 DuCV 的完全的染色体的四重叠碎片。特定的聚合酶链反应产品从积极样品被放大。碎片然后被克隆进 pMD18-T 向量并且定序，并且 DuCV 的 FJ0601 isolate 的完整的长度染色体组的顺序被获得。聚合酶链反应分析证明积极因为 circovirus 从 12 个农场是 79% 和 10 的鸭的比例是积极的。顺序分析证明 DuCV-FJ0601 的完全的染色体是 1988 bp 并且拥有了对包括了茎环结构和议员蛋白质主题的家庭 Circoviridae 普通的特征。相同分析证明 DuCV 的那 FJ0601 isolate 有到所有四个台湾的身份孤立的 97.3%&#8764;97.5% 核苷酸序列(TC1/2002， TC2/2002， TC3/2002， TC4/2002 ) ，到美洲(33753-52 )isolate 的 82.9% 身份和到德国的 82.3% 身份孤立。然而，有 Clustal W 的种系发生的分析证明 DuCV 的那 FJ0601 isolate 在有台湾的一个普通分支上孤立，并且德国和美洲孤立属于另外的分支。

The effects of Shuxuetong on the pathology of cerebral ischemia-reperfusion injury and GABA and TNF-α expression in gerbil models

Objective To study the protective effect and mechanism of Shuxuetong on gerbil brain tissue from the area of ischemia-reperfusion. Methods Cerebral ischemia-reperfusion animal model by transient clipping bilateral common carotid arteries in gerbils. Pathological changes in the hippocampal tissue were observed at different reperfusion time (12h, 3d, 7d). The expression levels of GABA and TNF-α in the hippocampal CA_1 subfield were observed using immunohistochemitry at 12 h, 3 d after reperfusion. The difference of above indices among false operation group, ischemia-reperfusion group and treatment group were compared. Results The injuries of pathology to hippocampal area in isehemiareperfusion group were more serious than treatment group. The expression levels of GABA in treatment group were significantly increased compared with ischemia-reperfusion group, but the expression levels of TNF-α between the two groups have no difference. Conclusion Shuxuetong has protective effect on brain tissue of ischemia-reperfusion by enhancing the expression of GABA in the hippocampal tissue.

Effects of Salvia miltiorrhiza Bge.f.alba on neuronal regeneration following cerebral ischemia/reperfusion

BACKGROUND： Subsequent to cerebral ischemic injury, endogenous neural stem cells are activated, but ischemia-induced neuronal loss is not compensated by ischemic injury-induced neural regeneration. Salvia （S.） miltiorrhiza Bge.f.alba （Baihua Danshen, a Chinese herbal medicine） could enhance learning and memory functions, as well as promote neural regeneration. OBJECTIVE： To observe the effects of S. miltiorrhiza Bge.f.alba on recovery from cerebral ischemia-reperfusion injury, and the influence on neuronal regeneration and differentiation. DESIGN, TIME AND SETTING： Randomized, controlled, animal experiments were performed at the Experimental Animal Center and Neurobiology Laboratory of Taishan Medical College in September of 2006. MATERIALS： S. miltiorrhiza Bge.f.alba was provided by Taishan Medical College Botanic Garden, Taian, China; dl-3n-butylphthalide （NBP） soft capsule was purchased from NBP Pharmaceutical, Shijiazhuang, China; mouse anti-bromodeoxyuridine antibody, rabbit anti-NF200 antibody, and bromodeoxyuridine were purchased from Sigma, Louis, MO, USA; Annexin V-fluorescein isothiocyanate/PI apoptosis kit was purchased from Nanjing Comissariado Biological Technology Development, Nanjing, China. METHODS： Adult Sprague Dawley rats were randomly assigned to sham surgery, model （cerebral ischemia and reperfusion, without administration）, S. miltiorrhiza Bge.f.alba, and NBP groups. Following establishment of the cerebral ischemia/reperfusion model, S. miltiorrhiza Bge.f.alba or NBP （1 mL/100 g） was respectively perfused at 30 minutes following cerebral ischemia/reperfusion. MAIN OUTCOME MEASURES： Alterations in cerebral blood flow before and after ischemia/reperfusion, NF200- and bromodeoxyuridine-double positive cells in striatum of affected tissues, as well as neuronal apoptosis rate at days 5 and 7 following cerebral ischemia/reperfusion. RESULTS： Subsequent to cerebral ischemia reperfusion, cerebral blood flow was reduced. Following treatment with S. miltiorrhiza Bge.f.alba, cerebral blood flow significantly increased （P 〈 0.05）. NBP treatment was inferior to S. miltiorrhiza Bge.f.alba with regard to stabilization of cerebral blood flow （P 〈 0.05）. S. miltiorrhiza Bge.f.alba significantly increased the number of newly formed neurons in rats following cerebral ischemia （P 〈 0.05） and significantly reduced neuronal apoptosis （P 〈 0.05）, with no significant difference compared with NBP treatment （P 〉 0.05）. CONCLUSION： S. miltiorrhiza Bge.f.alba significantly increased cerebral blood flow, reduced neuronal apoptosis, and promoted neuronal regeneration in rats with cerebral ischemia/reperfusion impairment.

Synergistic effects of ginseng stem and leaf-extracted ginsenoside and choline on improving learning and memory in rats Association verification experiment in animals with multiple learning and memory disorders

BACKGROUND： Ginsenoside extracted from the stem and leaf of ginseng （GSL） and choline have both been shown to improve learning and memory functions; however, further studies are needed to understand the synergistic effects of a combination of both. OBJECTIVE： To verify the combined improved synergistic effects of GSL and choline on learning and memory disorders in rats. DESIGN： Control observation. SETTING： Taishan Medical College. MATERIALS： A total of 150 male Kunming mice weighing （204-2） g and 40 healthy male Wistar rats weighing （2204-20） g were provided by the Experimental Animal Department of Jilin University. Animal experimentation received confirmed consent from the local ethic committee. GSL was provided by the Department of Chemistry, Norman Bethune Medical University, and choline was provided by the Third Experiment Factory, Shanghai. METHODS： This study was performed at the Life Science Institute, Taishan Medical College from October 2006 to February 2007. ① Scopolamine-induced learning and memory disorders in rats： Forty rats were randomly divided into control group, model group, combination group （400 mg/kg GSL ＋ 200 mg/kg choline）, GSL （400 mg/kg） group, and choline （200 mg/kg） group, 8 rats/group. Rats were perfused and administrated in the morning, once a day for 14 successive days. Rats in the control group and model group were perfused with 20 mL/kg distilled water and underwent Morris water maze spatial resolution test 1 hour after perfusion on the 10m, 11m, and 12m days after administration. Rats also underwent passive step-down avoidance test 1 hour after reperfusion on the 13m and 14m days after administration. Thirty minutes prior to experimentation, rats in the remaining three groups were intraperitoneally （i.p） injected with 2 mg/kg scopolamine, and rats in the control group were i.p. injected with 2 mL/kg saline. ② Scopolamine-induced learning disorder and memory acquired disorder in mice： Fifty mice were randomly divided into control group, model group, combination group （400 mg/kg GSL ＋200 mg/kg choline）, GSL （400 mg/kg） group, and choline （200 mg/kg） group, with 10 mice/group. Mice were perfused and administrated in the morning, once a day for 9 successive days. Mice in the control group and model group were perfused with 20 mL/kg distilled water and underwent passive step down avoidance test 1 hour after reperfusion on the 8th and 9th day after administration. Twenty minutes prior to training, mice in the remaining three groups were i.p. injected with 2 mg/kg scopolamine, and mice in the control group were i.p. injected with 10 mL/kg saline. ③ Sodium nitrite-induced memory consolidation disorder in mice： Grouping, administration, and testing were the same as mentioned above. After training, mice in the remaining three groups were immediately subcutaneously injected with 120 mg/kg sodium nitrite, and mice in the control group were subcutaneously injected with 20 mL/kg saline. ④ Ethanol-induced memory reconsolidation disorder in mice： Grouping, administration, and testing were the same as mentioned above. At 24 hours after training and 20 minutes before retraining, mice in the remaining four groups were perfused with 10 mL/kg ethanol （0.3 volume fraction）, and mice in the control group were perfused with 10 mL/kg saline. MAIN OUTCOME MEASURES： Synergistic effects of GSL and choline on learning and memory deficits induced by scopolamine, sodium nitrite, and ethanol in experimental animals. RESULTS： All 40 rats and 150 mice were included in the final analysis. ① Synergistic effects of GSL and choline on learning and memory disorders induced by scopolamine in rats： During passive step-down avoidance and Morris water maze spatial resolution tests, the number of error responses and length of maze training in the model group were significantly greater than in the control group （P 〈 0.01）; while the number of error responses and length of maze training in the combination group were significantly less than in the model group, GSL group, and choline group （P 〈 0.05-0.01）. The Q value was 〉 1 after combining administration, which suggests that the combination of GSL and choline had synergistic effects. ② Synergistic effects of GSL and choline on learning disorder and memory-acquired disorder induced by scopolamine in mice： During passive step-down avoidance test, the number of error responses in the model group were significantly greater than in the control group （P 〈 0.01 ）; while the number of error responses in the combination group were significantly less than in the model group, GSL group, and choline group （P 〈 0.05-0.01）. The Q value was 〉 1 after combining administration, which suggests GSL and choline had synergistic effects. ③ Synergistic effects of GSL and choline on memory sodium nitrate-induced consolidation disorder in mice： During passive step down avoidance test, the number of error responses in the model group were significantly less than in the control group （P 〈 0.01 ）; while the number of error responses in the combination group were significantly less than in the model group, GSL group, and choline group （P 〈 0.05-0.01）. The Q value was 〉 1 after combined administration, which suggests GSL and choline had synergistic effects. ④ Synergistic effects of GSL and choline on ethanol-induced memory reconsolidation disorder in mice： During passive step down avoidance test, the number of error responses in the model group were significantly greater than in the control group （P 〈 0.01）; while the number of error responses in the combination group were significantly less than in the model group, GSL group, and choline group （P 〈 0.05-0.01）. The Q value was 〉 1 after combined administration, which suggests GSL and choline had synergistic effects. CONCLUSION： GSL and choline have synergistic effects on learning and memory functions.

The Extraction of Eucommia Ulmoides Oliv Polysaccharides and Its Protective Effect on Liver of Clophasphamidecy Injured Mice

Objective:To study the influenceof eucommia polysaccharide on the mice' liver damaged by clophasphamidecy (CY).Methods:Injecting CY build mice liver damage model,eucommia polysaccharide given different doses, measured blood serum ALT,AST and the liver's SOD,MDA. Results:After the injection CY,blood serum ALT,AST and the MDA of liver rise and the SOD of liver reduce comparedwith the blank group. The eucommia polysaccharide can improve these index.Conclusion:The Eucommia polysaccharide may protect the mice' liver damaged by CY.

CLINICAL OBSERVATION ON THE TREATMENT OF CHRONIC FATIGUE SYNDROME WITH DEEPLY PUNCTURED ELONGATED NEEDLE COMBINED WITH COMPREHENSIVE NURSING

Objective: To observe the therapeutic effect of deep puncture of Zusanli (足三里ST 36) and Qihai (气海CV 6) with elongated needle combined with comprehensive nursing in the treatment of chronic fatigue syndrome (CFS) so as to provide new way and effective method for CFS. Methods: A total of 100 cases of CFS patients were evenly randomized into treatment group and control group which were treated with elongated needle puncture of ST 36 and CV 6 plus nursing and oral administration of Shiquan Dabu Tang (十全大补汤Decoction of Ten Powerful Tonics) separately. Acupuncture and medication were given once daily, with 7 sessions being a therapeutic course, 4 courses altogether. Results: After 4 courses of treatment, of the two 50 cases in treatment and control groups, 4 (8.0%) and 2 (4.0%) were cured, 37 (74.0%) and 16 (32.0%) experienced marked improvement, 6 (12.0%) and 19 (38.0%) had improvement, 3 (6.0%) and 13 (26.0%) failed, with the total effective rates being 94.0% (47/50) and 74.0% (37/50) respectively. The total effective rate and markedly effective rate of treatment group were significantly higher than those of control group (P<0.05, 0.01). Conclusion: Deep puncture of Zusanli (ST 36) and Qihai (CV 6) with elongated needle combined with comprehensive nursing has a definite therapeutic effect in the treatment of chronic fatigue syndrome.

Histological and ultrastructural changes induced by selenium in early experimental gastric carcinogenesis

AIM: To investigate the effect and significance of selenium in early experimental gastric carcinogenesis.METHODS: Weaning male Wistar rats were divided randomly into normal control group, experiment control group, low selenium (2 mg/L) group and high selenium (4 mg/L) group. Wistar rat gastric carcinogenesis was induced by N-methyl-N-nitro-N-nitroso guanidine (MNNG) (20 mg/kg) gavage daily for 10 d. Na2SeO3 was given by piped drinking 1 wk prior to MNNG gavage. The rats were killed at the 43rd wk. The surface characteristics of gastric mucosa were observed with naked eyes. Histopathologic changes of rat gastric mucosa were observed by HE staining and AB-PAS methods. The changes of cellular ultrastructure were observed under transmission electron microscope. Statistical analysis was carried out by SPSS.RESULTS: The incidence rate of gastric mucosa erosion,hemorrhage and intestinal metaplasia was 0, 45.5%,66.7%, and 92.9%, respectively (92.9% vs45.5%, P＜0.05)in the normal control group, experiment control group,low selenium group, and high selenium group. Leiomyoma formed in the process of inducement of rat gastric carcinoma. Dietary Na2SeO3 (2 and 4 mg/L) slightly increased the incidence rate of leiomyoma (0, 23%, 46.6%, and 46.6%). gastric mucosa did not change in the course of rat gastric carcinogenesis. Dietary Na2SeO3 by pipe drinking could expand the intracellular secretory canaliculus of parietal cells and increase the number of endocrine cells and lysosomes.CONCLUSION: Dietary Na2SeO3 by pipe drinking aggravates gastric erosion, hemorrhage and promotes intestinal metaplasia of gastric mucosa. The mechanism may be related with the function of parietal cells.

Construction of the Subtracted cDNA Library of Striatal Neurons Treated with Long-term Morphine

Objective To construct a morphine tolerance model in primarily cultured striatal neurons,and screen the differentially expressed genes in this model using suppression subtractive hybridization (SSH).Methods Sbtracted cDNA libraries were constructed using SSH from normal primarily cultured striatal neurons and long-term morphine treated striatal neurons (10-5 mol/L for 72 hours).To check reliability of the cell culture model,RT-PCR was performed to detect the cAMP-responsive element-binding protein (CREB) mRNA expression.The subtracted clones were prescreened by PCR.The clones containing inserted fragments from forward libraries were sequenced and submitted to GenBank for homology analysis.And the expression levels of genes of interest were confirmed by RT-PCR.Results CREB mRNA expression showed a significant increase in morphine treated striatal neurons (62.85±1.98) compared with normal striatal neurons (28.43±1.46,P<0.01).Thirty-six clones containing inserted fragments were randomly chosen for sequence analysis.And the 36 clones showed homology with 19 known genes and 2 novel genes.The expression of 2 novel genes,mitochondrial carrier homolog 1 (Mtch1;96.81±2.04 vs.44.20±1.31,P<0.01) and thymoma viral proto-oncogene 1 (Akt1;122.10±2.17 vs.50.11±2.01,P<0.01),showed a significant increase in morphine-treated striatal neurons compared with normal striatal neurons.Conclusions A reliable differential cDNA library of striatal neurons treated with long-term morphine is constructed.Mtch1 and Akt1 might be the candidate genes for the development of morphine tolerance.

Elevated Plasma Tissue-type Plasminogen Activator (t-PA) and Soluble Throm-bomodulin in Patients Suffering From Severe Acute Respiratory Syndrome (SARS) as a Possible Index for Prognosis and Treatment Strategy

To detect the presence of endothelial injury in patients with severe acute respiratory syndrome （SARS） via enhanced levels of tissue-type plasminogen activator （t-PA） and soluble thrombomodulin （sTM）. Methods Case patients were from Xuanwu Hospital （Capital University of Medical Sciences, Beijing, China）, and all of them met clinical criteria for SARS. Healthy controls were some of the hospital employees. Endothelial injury bio-markers tPA and sTM were detected by commercial ELISA-methods. Results Classic plasma markers of endothelial injury, tPA and sTM significantly elevated in SARS patients in comparison to controls [t-PA： 1.48±0.16 nmol/L versus 0.25±0.03 nmol/L （P〈0.0001）, and sTM： 0.26±0.06 nmol/L versus 0.14±0.02 nmol/L （P〈0.05）]. The only patient who died had extremely high levels of these endothelial injury markers （t-PA： 2.77 nmol/L and sTM： 1.01 nmol/L）. The likelihood ratio analysis indicated the excellent discriminating power for SARS at the optimal cut-point of 0.49 nmol/L for tPA and 0.20 nmol/L for sTM, respectively. Significant numerical correlations were found among these endothelial injury markers in SARS patients. The numerical coefficient of correlation Pearson r between t-PA and sTM was 0.5867 （P〈0.05）. Conclusion Increased plasma concentrations of tPA and sTM in patients with SARS suggest the possibility of endothelial injury. SARS patients might need anticoagulant therapy or fibrinolytic therapy in order to reverse intraalveolar coagulation, microthrombi formation, alveolar and interstitial fibrin deposition. It may not only provide a useful treatment and prognostic index but also allow a further understanding of the pathological condition of the disease.

An erratum on “Application of three-dimensional printing in interventional medicine” [J Interv Med(February 2020) 1–16]

After the publication of this work,1 the authors noticed and confirmed that the Funding Information was mistakenly omitted from the article.The statement“This study was supported by grants from the National Natural Science Foundation of China,grant no.81370041,81471760,81671655,the Outstanding Clinical Discipline Project of Shanghai Pudong,grant no.PWYgy2018-04.The authors declare that they have no conflicts of interest.”should be included in the Funding information section of the paper which is missing.We apologize for the error.

Protective effects of a polysaccharide from Spirulina platensis on dopaminergic neurons in an MPT-Pinduced Parkinson's disease model in C57BL/6J mice

The present study aimed to determine whether a polysaccharide obtained fromSpirulina platensis shows protective effects on dopaminergic neurons. A Parkinson’s disease model was established through the intraperitoneal injection of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyr-idine （MPTP） in C57BL/6J mice. Prior to the MPTP injection, some mice were pretreated with intraperitoneal injections of a polysaccharide derived fromSpirulina platensis once daily for 10 days. The results showed that the immunoreactive staining and mRNA expression of the dopa-mine transporter and tyrosine hydroxylase, the rate-limiting enzyme in dopamine synthesis, in the substantia nigra, were signiifcantly increased in mice pretreated with 800 mg/kg of the poly-saccharide compared with those in MPTP-treated mice. The activities of superoxide dismutase and glutathione peroxidase in the serum and midbrain were also increased signiifcantly in mice injected with MPTP after pretreatment with the polysaccharide fromSpirulina platensis. By con-trast, the activity of monoamine oxidase B in serum and midbrain maintained unchanged. These experimental ifndings indicate that the polysaccharide obtained fromSpirulina platensis plays a protective role against the MPTP-induced loss of dopaminergic neurons in C57BL/6J mice, and that the antioxidative properties of this polysaccharide likely underlie its neuroprotective effect.

Selection Pressure on Haemagglutinin Genes of H9N2 Influenza Viruses from Different Hosts

Positive selection and differential selective pressure analyses were carried out to study Haemagglutinin (HA) genes of H9N2 influenza viruses from different hosts in this paper. Results showed that, although most positions in HAs were under neutral or purifying evolution, a few positions located in the antigenic regions and receptor binding sites were subject to positive selection and some of them were even positively selected at the population level. In addition, there were always some positions differentially selected for viruses from different hosts. Both selection pressure working on HA codons and positions differentially selected might account for the extension of the host range and adaptations to different hosts of H9N2 influenza viruses.

Simultaneous detection of duck hepatitis A virus types 1 and 3, and of duck astrovirus type 1, by multiplex RT-PCR

Dear Editor,Duck virus hepatitis(DVH)is caused by at least threedifferent RNA viruses,including duck hepatitis A virus(DHAV),duck astrovirus type 1(DAstV-1),and duckastrovirus type 2(DAstV-2).The first of these,DHAV,has been classified into three serotypes by

Potential risk of mitomycin C at high concentrations on peripheral nerve structure

Although the local application of mitomycin C may prevent epidural adhesion after laminectomy, mitomycin C can induce neurotoxicity in optic and acoustic nerves at high concentrations. To determine the safe concentration range for mitomycin C, cotton pads soaked with mitomycin C at different concentrations （0.1, 0.3, 0.5, and 0.7 mg/mL） were immediately applied for 5 minutes to the operation area of rats that had undergone laminectomy at L1. Rat sciatic nerves, instead of dorsal nerves, were used in this study. The results showed that mitomycin C at 0.1-0.5 mg/mL did not damage the structure and function of the sciatic nerve, while at 0.7 mg/mL, mitomycin C signiifcantly reduced the thickness of the sciatic nerve myelin sheath compared with lower concen-trations, though no functional change was found. These experimental ifndings indicate that the local application of mitomycin C at low concentrations is safe to prevent scar adhesion following laminectomy, but that mitomycin C at high concentrations （＆gt;0.7 mg/mL） has potential safety risks to peripheral nerve structures.

CLINICAL OBSERVATION ON 96 CASES OF PRIMARY DYSMENORRHEA TREATED BY MEDICINE-SEPARATED MOXIBUSTION AND INVESTIGATION ON ITS MECHANISMS

Objective: To observe the therapeutic effect of medicine-separated moxibustion for primary dysmenorrhea and its influence on plasma hormone level. Methods: Ninety-six patients suffering from primary dysmenorrhea were randomly and evenly divided into treatment group treated with medicine-separated moxibustion of Shenque （神阙CV 8, 3-5 cones every time, beginning 1 week before onset of menstruation and stopping on the 3 rd day after onset, with 10 sessions being a therapeutic course, 3 courses all together）, and control group treated with oral administration of Yueyueshu （月月舒menstruation-smoothing granule, 10 g/time, b.i.d, 3 courses altogether）. Menses prostaglandin E2 （PGF 2α ） and plasma oxytocin （OT） during menstruation were determined by radioimmunoassay. Results: After the treatment, of the two 48 cases in treatment and control groups, 18 and 5 were cured, 24 and 9 had marked improvement in their symptoms, 6 and 26 had improvement, 0 and 8 failed in the treatment, with the total effective rates being 100.0% and 83.3% respectively, the therapeutic effect of treatment group was markedly superior to that of control group （P<0.05）. After the treatment, the contents of menses PGF 2α in treatment group and plasma OT in both groups were significantly lower than those of pre-treatment （P<0.01）. The therapeutic effect of moxibustion was significantly better than that of medication in lowering plasma OT. Conclusion: Medicine-separated moxibustion works well in treating primary dysmenorrhea, and moxibustion induced decrease of menses PGF 2α and plasma OT may contribute to its effect in relieving dysmenorrhea.