Objective To investigate the regulation of blood-testis barrier by Rab13-PKA pathway in rats.Meth-od First, shRNA vector targeting at Rab13 was constructed and then the Rab13 shRNA was transfected into the rat testis by injection.Western blot was used to detect the knock-down effect of Rab13 and the expression of blood-testis barrier ( BTB) constituent proteins.PKA activity was detected by autoradiography and scintillation counting.Further, immunoflu-orescence analysis and phalloidin staining were applied to observe the distribution of occludin and F-actin, respectively. Results The expression level of Rab13 in the testis was reduced by approximately 70%after transfection of Rab13 shRNA as compared with the non-targeted control group ( P<0.01 ) , while the expression of BTB constituent proteins remained unchanged.PKA activity was significantly increased after Rab13 RNAi transfection (P<0.01).The distribution of occlu-din at BTB was remarkably increased after Rab13 RNAi silencing around stage VIII but not at other stages of the seminifer-ous epithelial cycle.The assembly of F-actin at BTB was also intensified in Rab13-silenced testis.Both the changes of dis-tribution of occludin and F-actin induced by Rab13 shRNA were found to be antagonized by the PKA specific inhibitor H89.Conclusions Rab13 can modulate the distribution of occludin and F-actin at the blood-testis barrier in rats by regu-lating PKA activity, which may participate in the regulation of BTB function.

Background and purpose:This study investigated the relationship between (S100 calcium-binding protein A4, S100A4) in chronic gastritis, intestinal metaplasia, dysplasia adenomatous, normal tissue tissue samples and expression in gastric cancer and clinical characteristics. Methods:HE staining of the use of gastric specimens taken for histopathological diagnosis;using immunohistochemistry to detect the expression of tissue S100A4 protein;qRT-PCR was used to detect mRNA expression of S100A4 gene;Western Blot detection of S100A4 gene encoding protein. Kaplan-Meier survival curves were used to distinguish and compare survival. Results:S100A4 protein and mRNA expression gradually increased in the following order:normal tissue

Objective To study the clinical discrepancy between patients with post infectious irritable bowel syndrome(PI-IBS) and non post infectious irritable bowel syndrome(NPI-IBS),and assess the value of serum intestinal fatty acid binding protein (I-FABP) for differential diagnosis.Methods A total of 117 patients with PI-IBS,201 patients with NPI-IBS and 31 healthy controls were prospectively recruited in General Liberation Army Hospital from 2010 to 2013.Plasma samples and clinical data were collected.Serum I-FABP level was measured by an enzyme-linked immunosorbent assay.Results The median age of patients with PI-IBS was 36 years.The median time to diagnosis in PI-IBS group was significantly longer than that in NPI-IBS group [(19.7 ± 10.3) months vs (11.4 ± 5.3) months,P ＜ 0.05].Similarly,the proportion of anxiety [58.1％ (68/117) vs 28.9％ (58/201),P ＜ 0.05] and the value of I-FABP[(42.6 ± 14.8) μg/L vs (17.3 ± 11.5) μg/L,P ＜ 0.05] in PI-IBS group were significant higher than NPI-IBS patients.The level of I-FABP of healthy controls [(10.6 ± 8.2) μg/L] was also significantly lower than that of PI-IBS patients (P ＜ 0.05),yet no difference from that of NPI-IBS group.The I-FABP value of subgroup PI-IBS patients with diarrhoea (IBS-D) was significant higher than that of NPI-IBS group [(54.8 ± 9.3) μg/L vs (12.3 ± 6.2) μg/L,P ＜ 0.05].However,other parameters including gender,age,GSRS score,and I-FABP value of subgroup constipation (IBS-C) and mix (IBS-M),were not different between PI-IBS group and NPI-IBS group (all P ＞ 0.05).Conclusion PI-IBS is an occult intestinal inflammation disease with mucosa injury.I-FABP might be a potential testing marker for the diagnosis of PI-IBS.

Objective To retrospectively analyze and summarize the manipulation and skills of the placement of uaso-jejunal feeding tube under DSA guidance. Methods After performing the spraying anesthesia of nasopharynx, the naso-jejunal feeding tube, with the help of guide wire and under DSA guidance, was placed into the proximal jejunum by passing it through the nose, pharynx, esophagus, stomach,pylorus and duodenum in order. The procedure was employed in 441 cases. Results The mean time for performing the procedure was within five minutes. The procedure was successfully accomplished in all 441 cases and no complications occurred in this series. Conclusion Under DSA guidance the placement of nasojejunal feeding tube can be safely and quickly carried out with high successful rate and less sufferings to patient. It is worth popularizing this technique in clinical practice.

OBJECTIVE:To provide reference for further standardizing and perfecting the management of drug unpacking in outpatient pharmacy.METHODS:A total of 6 primary and secondary health institutions (4 community health service centers and 2 secondary hospitals) were selected from 2 districts in Shanghai to conduct a questionnaire survey on the use of their drugs and drug unpacking in outpatient pharmacy.The survey data was analyzed statistically.RESULTS:Totally 6 institution questionnaires and 6 pharmaceutical staff questionnaires were sent out,all were received with recovery of 100％.In 2015,the average number of essential medicines in community health service centers and secondary hospitals were 496.50,542.00,respectively,and the average number of varieties sold were 530.75,1 052.00.In outpatient pharmacy of surveyed community health service center,the number of unpacked drugs was 10-21 which were essential drugs and class A medical insurance drugs,and the consumption sum of unpacked drug ranged 5.56-16.70 ten thousand yuan.In outpatient pharmacy of 2 two secondary hospitals,the number of unpacked drugs were 17 and 23,respectively,most of which were essential drugs and class A medical insurance drugs,and the consumption sum of unpacked drug ranged 13.19 to 158.06 ten thousand yuan.The proportion of unpacked drugs was less than 5％ of the total number of varieties sold,and the proportion of consumption sum of unpacked drugs was less than 1％ of total consumption sum.Estazolam tablets and Alprazolam tablets took up the top 5 in the list of consumption sum of unpacking drugs in 2 types of intervi ewed instiutions.All the surveyed institutions were not equipped with drug dispensing machine in the outpatient pharmacy,still depended on manually unpacking.There were four institutions to regularly arrange the unpacking,unpacking frequency was usually 1 to 3 times a week,supplemented by the need to arrange unpacking,and another two to implement a daily unpacking.There were 5 institutions to develop a drug unpacking mechanism in the institutions,but the relevant system was not perfect.CONCLUSIONS:The enthusiasm of pharmaceutical saff in primary and secondary health institutions in Shanghai to carry out or engage in unpacking work need to be improved,the instructions for unpacked drugs are not available on request,and the way to unpack drugs still needs to explore.

ObjectiveTo assess the differential diagnostic value of serum intestinal fatty acid binding protein (I-FABP)in distinguishing intestinal ischemia patients from acute abdomen patients.MethodsA total of 151 patients with acute abdomen and 17 healthy controls from the PLA General Hospital were enrolled from November,2009 to August,2011. Serum I-FABP levels were measured by ELISA.According to the ROC curve,the cut-off value,sensitivity,specificity,positive likelihood ratio (PLR),negative likelihood ratio ( NLR),positive predietive value (PPV) and negative predictive value (NPV) were calculated. ResultsOf the 151 acute abdomen patients,there were 24 intestinal ischemia patients and 127 without intestinal ischemia.Serum I-FABP level in intestinal ischemia group [( 109.67 ±48.82) μg/L]was significantly higher than those in patients without intestinal ischemia [(36.78 ± 11.25) μg/L]and healthy controls[(8.33 ±6.25) μg/L]( all P values ＜0.01 ).The serum I-FABP cut-off value for the diagnosis of intestinal ischemia was 87.52 μg/L.Serum I-FABP was efficient in terms of sensitivity (0.762),NPV(0.963),PLR(3.05) and NLR (0.24) in the diagnosis of intestinal ischemia.ConclusionI-FABP is potentially useful for discriminating intestinal ischemia from acute abdomen.

Objective To establish an HPLC method for the determination of anthraquinones including rhein, emodin and chrysophanol in xinshenyan capsules. Methods Anthraquinones were determined by HPLC with Phenomenex-C18 column (250 mm×4. 6 mm, 5 μm) as the chromatographic column and methanol-1% acetic acid (70:30) as the mobile phase. The flow rate was 1. 0 mL·min-1 and the detection wavelength was set at 254 nm. Results The liner range of rhein, emodin and chrysophanol was 4. 96-24. 80 μg·mL-1(r=0. 999 6), 6. 58-32. 91 μg·mL-1(r=0. 999 9) ,and 15. 11-75. 55 μg·mL-1 (r=0. 999 9),respectively, and the average recovery was 100. 78%, 98. 13% and 99. 29%, respectively. Conclusion The method is simple and practical, the result is accurate and reliable and it can be used to determine the contents of rhein, emodin and chrysophanol in xinshenyan capsules.

Objective:To establish the quality control of Zhiqikang capsules. Methods:TLC was used to identify Gastrodia tuder halimasch, rhubarb and Astragalus mongholicus in the preparations. A spectrophotometry method with 3, 5-dinitrosalicylic acid (DNS) was used to measure the polysaccharide content in Zhiqikang capsules. A spectrophotometry method with Forint phenol method ( Low-ry) was used to measure the peptide content in the capsules. Results:The linear range of polysaccharide was obtained between 6. 412 and 32. 060μg·ml-1(r=0. 999 5), the average recovery was 95. 86% and RSD was 0. 86%. The linear range of peptide was ob-tained between 0.059 7 and 0.298 4 mg·ml-1(r=0.999 0), the average recovery was 100.3% and RSD was 1.88%(n=6). Conclusion:The assay method is simple and accurate in the quality control of the preparations.

To investigate the function of testis sperm binding protein (TSBP) in sperm capacitation and acrosome reaction, the effect of the recombinant TSBP on the activity of protein kinase A was detected in the transfected cell line. With the use of prokaryotic expressing plasmid pGEX-5X-1/tsbp as template, the novel gene tsbp was amplified by PCR and a eukaryotic expressing vector pcDNA3.1/myc-His(-)B/tsbp was constructed. DNA sequencing and restriction endonuclease digestion analysis indicated that the eukaryotic expression vector pcDNA3.1/myc-His(-)B/tsbp had been constructed successfully. After the recombinant plamid being transfected into HEK293 cells, Western blot verified the expression of tsbp. Fusion protein His6-TSBP was purified from the cell lysate by immobilized metal-ion affinity chromatography (IMAC). Radioautograph revealed a higher PKA activity in the transfected HEK293 cells than in the control group, which indicates that TSBP can increase the activity of PKA.
Acrosome Reaction ; drug effects ; Cell Line ; Cyclic AMP-Dependent Protein Kinases ; drug effects ; Humans ; Male ; Recombinant Proteins ; biosynthesis ; genetics ; pharmacology ; Seminal Plasma Proteins ; biosynthesis ; genetics ; pharmacology ; Sperm Capacitation ; drug effects

Response surface methodology was applied to optimize the fermentation conditions of FGFC1 (Fungi fibrinolytic compound 1). On the basis of single factor tests, response surface analysis was designed by Design-Expert, and the effects of culture time, ornithine hydrochloride addition and culture temperature on the yield of FGFC1 were studied, the predicted value and measured value were also contrasted. The results show the optimal culture conditions as follows: the culture time is 7 d, ornithine hydrochloride addition is 0.5% (M/V), culture temperature is 28 degrees C. Under these conditions, the yield of FGFC1 is 1 978.33 mg/L, which is consistent with the predicted value. It shows that the experiment is effective.
Culture Techniques ; methods ; Fermentation ; Fibrinolytic Agents ; metabolism ; Seawater ; microbiology ; Stachybotrys ; growth & development ; metabolism ; Surface Properties