AIM: To establish the qualitative method for determining hirudo in Zhiyu Granule (Scorpio, Scolopendra, Rhizoma Coptidis, Radix Astragali, etc.) METHODS: Hirudo was determined by anti-thrombin bioactivity method after being extracted from Zhiyu Granule using 95% ethanol. RESULTS: The difference between Zhiyu Granule and the control sample without hirude was significant and easy to identify. CONCLUSION: After being extracted by 95% ethanol, hirudo can be identified from other anti-thrombin bioactivity components in prescription.

This paper elaborated the research progress on transduction mechanisms of obese polycystic ovary syndrome (PCOS) and insulin resistance (IR). The action mechanisms of kidney-tonifying, spleen-invigorating and phlegm-resolving in the treatment of obese PCOS and IR were explored. It provided new ideas for the treatment of obese PCOS patients in the clinical practice of traditional Chinese medicine (TCM) gynecology. Literatures on TCM theories and modern signal pathways were used in the analysis of spleen-kidney deficiency and phlegm-dampness retention, which were the key pathogenesis of obese PCOS and IR. The scientific nature of treating obese PCOS and IR from the method of kidney-tonifying, spleen-invigorating and phlegm-resolving was demonstrated. The results showed that the scientific nature of treating obese PCOS and IR from the method of kidney-tonifying, spleen-invigorating and phlegm-resolving was initially demonstrated by the organic combination of TCM and modern medicine theories. It innovatively proposed that the treatment study strategy of transduction molecular mechanism took the interactive dialogue between PI-3K/Akt signal pathway and TNF-α signal pathway as its target. The regulatory role and action mechanism of this method in the treatment of obese PCOS and IR were discussed.

This study was aimed to establish and evaluate obese polycystic ovary syndrome (PCOS) rat model. Rats were randomly divided into the blank control group, high-fat model group, insulin (INS) combined human chorionic gonadotropin (HCG) model group, and INS combined HCG plus high fat emulsion model group. The obese PCOS rat model was induced by subcutaneous injection of INS and HCG on the nape, respectively. The intragastric administration of high fat emulsion was also used in the PCOS rat model establishment. The estrous cycle of rat was monitored. The detection was also made the weight increasing rate of rats, luteinizing hormone (LH), follicle-stimulating hormone (FSH), estradiol (E2), testosterone (T), high-density lipoprotein (HDL), low density lipoprotein (LDL), triglyceride (TG), cholesterol (TC), fasting plasma glucose (FPG), fasting insulin (FINS), 2 h blood glucose and 2 h INS. Calculation was made on the HOMA index, related viscera index and bilateral ovaries HE staining. The results showed that when INS combined HCG improved method (INS combined HCG plus high fat emulsion model group) rats were sacrificed, the body weight, weight increasing rate, ovary viscera index, T, LH, and HOMA index were significantly higher than that of the blank control group, high-fat model group and INS combined HCG model group (P < 0.05,P < 0.01); HDL was significantly decreased (P < 0.05,P < 0.01). FPG, FINS, and 2 h INS of the INS combined HCG improved method group were significantly higher than that of the high-fat model group (P <0.01). It was concluded that INS combined HCG improved method was one of the ideal animal model establishment methods in the pathogenesis study of PCOS.

Objective To investigate the relationship of TNF-αgene promoter 308 locus(TNF-α-308)polymorphism with the susceptibility and severity of central venous catheter-related sepsis(CRS). Methods One hundred and five CRS patients admitted in Kaihua People's Hospital from January 2015 to May 2017 were enrolled in the study.According to whether complicated with multiple organ dysfunction syndrome(MODS), they were divided into CRS complicated MODS group(n=34)and CRS non-MODS group(n=71).Meanwhile,210 patients with no catheter-related infection(case control group)and 105 healthy subjects(healthy control group)were also enrolled in the study.Polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP)was used to genotype TNF-α-308, and the relationship of TNF-α-308 polymorphism with the susceptibility and severity of CRS was investigated.SPSS 16.0 was used to analyze the data.Results There were no significant differences in frequencies of GG, GA,AA genotypes and G,A allele of TNF-α-308 among CRS group,case control group and healthy control group(χ2=2.262 and 0.907,both P>0.05).Compared with CRS non-MODS group,case control group and healthy control group, the frequency of GG genotype was significantly lower and the frequencies of genotype GA and AA of TNF-α-308 were significantly higher in CRS MODS group(χ2=8.809,7.700 and 9.220,all P<0.05).Compared with CRS non-MODS group,case control group and healthy control group, the allele frequencies of G were significantly lower and allele frequency of A allele of TNF-α-308 was significantly higher in CRS MODS group(χ2=9.823, 8.624 and 7.654, all P<0.05).There were no significant differences in genotype frequency and allele frequencies of TNF-α-308(χ2=0.852 and 0.975, both P>0.05)among CRS non-MODS group and case control group,healthy control group(χ2=1.022 and 0.535,both P>0.05).The odds ratio of GA +AA genotype and A allele of TNF-α-308 in CRS MODS group were 2.664(95%CI 1.259-5.639)and 2.440(95%CI 1.326-4.490).Conclusion TNF-αgene promoter 308 locus polymorphism is not a predisposing factor for CRS, but may be associated with complication of MODS in CRS patients.

Objective To study the mechanism of Yangjing Zhongyu Decoction in regulating the initiation of primordial follicles in model rats with diminished ovarian reserve(DOR)based on lncRNA.Methods Three-day-old female rats were selected and their ovaries were cultured in vitro.The blank group,model group,DHEA group and Yangjing Zhongyu Decoction high-,medium-and low-dosage groups were set.The DOR model was induced by triptolide,corresponding drug containing serum was given to culture respectively.HE staining was used to observe germ cells and follicles,Western blot was used for determining the expressions of AMH,BMP15,PTEN,MST,TGF-β1,p-Smad1 protein,RT-PCR was used to detected AMH,BMP15,PTEN,MST,LTCONS-00011173,TGF-β1,Smad1 mRNA expression.Results Compared with the blank group,the number of primordial and growing follicles in the model group rats decreased(P<0.05),the expression of AMH,BMP15,TGF-β1,p-Smad1 protein in ovarian tissue decreased(P<0.05),expressions of PTEN and MST proteins increased(P<0.05),AMH,BMP15,TGF-β1,Smad1 mRNA expression decreased(P<0.05),while the expressions of PTEN,MST,and LTCONS-00011173 mRNA increased(P<0.05).Compared with the model group,the DHEA group and Yangjing Zhongyu Decoction high-and medium-dosage groups showed an increase in the number of primordial and growing follicles(P<0.05),the expressions of AMH,BMP15,TGF-β1 and p-Smad1 protein in ovarian tissue increased(P<0.05),PTEN and MST protein expressions decreased(P<0.05),AMH,BMP15,TGF-β1,Smad1 mRNA expressions increased(P<0.05),while PTEN,MST,and LTCONS-00011173 mRNA expressions decreased(P<0.05).Conclusion Yangjing Zhongyu Decoction may mediate TGF-β1/Smad1 signaling pathway through LTCONS-00011173,regulating primordial follicle initiation in DOR model rats.

Objective To investigate the effect of lower thoracic epidural block on intestinal epithelial cell apoptosis during hemorrhagic shock and resuscitation in rats.Methods Sixty-four male SD rats placed with lower thoracic epidural catheter were randomly divided into four groups (n=1 6 each):group Sham (sham operation),group HSR (hemorrhagic shock and resuscitation),group NS (hemorrhagic shock and resuscitation+epidural saline 100 μl/kg),and group TEA (hemorrhagic shock and resuscitation+epidural 0.075% ropivacaine 100 μl/kg).The hemorrhagic shock was made described by Chaudry.Rats were resuscitated by transfusing shed blood and normal saline 60 min after hemorrhagic shock.Malondialdehyde (MDA)content,superoxide dismutase (SOD)activity, and protein expression of Bax and Bcl-2 in intestinal epithelium were detected,and epithelial apoptosis index was calculated at 2 h after resuscitation.Results Compared with group Sham,intestinal epithe-lial MDA,Bax expression and epithelial apoptosis were significantly increased,while SOD activity were markedly decreased in groups HSR,NS and TEA (P<0.05).Compared with groups HSR and NS,intestinal epithelial MDA,Bax expression and epithelial apoptosis were significantly decreased, while SOD activity and Bcl-2 expression were markedly increased in group TEA (P <0.05 ). Conclusion Lower thoracic epidural block can enhance the antioxidant and anti-apoptotic ability,and inhibit the oxidative stress and cell apoptosis of intestinal epithelium.Therefore,it can promote the survival rate after hemorrhagic shock and resuscitation through protecting intestinal barrier.