Objective To evaluate the role of transient receptor potential ankyrin 1 (TRPA1) in the dorsal root ganglion neurons in the development of diabetic neuropathic pain (DNP) in rats.Methods Twenty-four Sprague-Dawley rats with DNP were randomly divided into 3 groups (n-=8 each) using a random number table:DNP group,TRPA1-specific siRNA group (siRNA group) and TRPA1-negative siRNA group (NC group).Another 8 Sprague-Dawley rats with normal blood glucose served as control group (C group).In siRNA group,TRPA1-specific siRNA 45 μl was injected intrathecally.In NC group,TRPA1-negative siRNA 45 μl was injected intrathecally.In DNP and C groups,normal saline 45 μl was injected intrathecally.On 2nd day after intrathecal administration,the lumbar segment (L4-6) of the dorsal root ganglions was removed for determination of the expression of TRPA1 mRNA.On 7,14,21 and 28 days after intrathecal administration (T1-4),MWT was measured.Results Compared with DNP group,TRPA1 mRNA expression was down-regulated in siRNA and C groups.Compared with DNP group,and MWT was significantly decreased at T1.2 in siRNA group,MWT was decreased at T1-3 in NC group,MWT was increased at T1-4 in group C.Compared with siRNA group,MWT was significantly increased at T1-4 in group C.MWT was significantly higher at T1~ in group C than in NC group.Conclusion TRPA1 in the dorsal root ganglion neurons is involved in the development of DNP in rats.

BACKGROUND:In the cytotoxicity test of the mussel adhesive protein dressing for wound healing, because of positive charge properties of the protein, when extracting ratio is 1:9, the cel s exhibit poly-group phenomenon that results in errors in the cytotoxicity test of mussel adhesive protein samples.
OBJECTIVE:According to the existing standards, to improve the leaching proportion and pretreatment of mussel adhesive protein dressing for wound healing based on the special properties and working condition of mussel adhesive protein.
METHODS:(1) Extract method:Extract solution of mussel adhesive protein dressing was prepared with mussel adhesive protein dressing and cel medium at extracting ratios of 1:9 and 1:131. Then, L929 cel s were cultured in extract solutions of mussel adhesive protein dressing, natural latex and high-density polyethylene, respectively. (2) Method of direct contact:Distil ed water, solutions of mussel adhesive protein dressing, dimethyl sulfoxide and cel medium were used to culture L929 cel s.
RESULTS AND CONCLUSION:When the extracting ratio was 1:9, the cel s agglomerated which is not suitable for cytotoxicity test. When the extracting ratio was 1:131, flocculated sediment and cel aggregation disappeared, the cytotoxicity test results showed no cytotoxicity with higher reliability. Direct contact method showed the samples had no cytotoxicity. The extract method with adjusted extracting radio or direct contact method can be applied to test the in vitro cytotoxicity of mussel adhesive protein dressing for wound healing.

To explore the operation methods and indications of the duodenoscopic papillotomy (IEST) with endoscopic nasobiliary drainage (IENBD) for the treatment of duodenal papilla stenosis during the course of common bile duct operation.The clinical data of 219 cases of cholecystolithiasis with choledocholith and the stenosis of papillary underwent endoscopic sphincterotomy (IEST) plus endoscopic nasobiliary drainage (IENBD) in the Second People's Hospital of Chengdu were retrospectively analyzed.It was successful in 198 cases who had the gallbladder and common bile duct stones removed,and endoscopic papillary dissection was performed and the nasobiliary tube was successfully inserted.Nasobiliary drainage was successful in 186 cases (93.9％) of 198 cases.No liquid outflow was observed in nasobiliary drainage in 7 cases (3.5％).Nasal bile duct slipped early in 5 case (2.5％).Primary closure of bile duct incision was completed in 198 cases.It failed in 4 cases (2.0％) who had the bile leakage with primary closure of duct incision.Mild pancreatitis after operation occurred in 3 cases (1.5％).Nose bile duct ligation was performed in 1 case (0.5％).The overall postoperative complication rate was 4.0％ (8/198).IEST + IENBD in open laparotomy was successful in 21 cases.No perforation of intestine and bile duct,bleeding,severe pancreatitis and other complications and death were detected postoperatively in two groups.During the course of laparoscopy and open laparotomy,IEST + IENBD in treating cholecystolithiasis with choledocholith and the stenosis of papillary and primary closure of duct incision after the endoscopic nasobiliary drainage is safe and effective.

Objective To evaluate laparoscopic balloon nasobiliary biliary drainage (LBNBD),vs ureteral catheter drainage in one stage laparoscopy,choledochoscopy and duodenoscopy choledocholithotomy and primary closure of the small calibered common bile duct (diameter 0.3-0.8 cm).Methods During the period of Apr 2010 to Nov 2016 102 cases were enrolled including 50 cases receiving LBNBD and 52 cases using ureteral catheter drainage.Results Between the two groups,LBNBD was superior to ureteral catheter drainage in all the following parameters:the operation time,intraoperative blood loss,postoperative liver function,blood amylase and other laboratory indicators,gastrointestinal function recovery time,gastrointestinal symptoms and electrolyte imbalance,postoperative hospital stay,and bile duct drainage time with all differences statistically significant (P ＜ 0.05).Bile drainage differences during the postoperative first 3 days (averagely 200-400 ml a day) were not statistically different (P ＞ 0.05).Postoperative pancreatitis,bile leakage,and hemobilia were not statistically different (P ＞ 0.05).Conclusions The use of LBNBD is safe and effective in endoscopic choledocholithotomy in cases of small calibered common bile duct.

Objective To modify the purified water analysis methods in the Chinese Pharmacopoeia (Vol .Ⅱ ,Edition 2010) ,and mainly focus on the pH value determination ,analysis of nitrate and nitrite ,and the inspection of microorganism . Methods The usage of regents and colorimetric tubes in the analysis of nitrate and nitrite were inquired ,the feasibility of plat-ing culture used in the inspection of microorganism were proposed ,and the suggestion of using more instrumental analysis in-stead of titrimetric analysis in the purified water inspection wasraised .Results The choice of colormetric apparatus and the precision of the regents were explicit .The plating method was better in convenience and reliability than membrane-filter proce-dure during the microorganism inspection .Instrumental analysis is the trend of purified water inspection and deserved to be rec-ommended .Conclusion Based on our own and others′studies ,the methods of impurity analysis are improved ,which might be accepted in the revision of Chinese Pharmacopoeia in future .

Objetive： To investigate the mechanism of antidepressant effect of verbascoside based on high-throughput sequencing technology （RNA-Seq），and to explore the possible targets and signaling pathways. MethodsForty C57BL/6 mice were randomly divided into control group，model group，fluoxetine group and verbascoside group，10 mice in each group. Except for the control group，all the other three groups were constructed with chronic unpredictable mild stimulation （CUMS） combined with solitary feeding for four weeks. Control group，fluoxetine group and verbascoside group were administered by gavage once daily for three weeks during the second week of modeling. The mice were assessed by sugar-water preference test，forced swimming test，open field test，elevated cross maze test，and water maze test. Enzyme linked immunosorbent assay（ELISA） was performed to detect the levels of major neurotransmitters and inflammatory factors in mice serum，and mRNA high-throughput sequencing was performed in the nucleus accumben and colon to screen differentially expressed genes and perform pathway enrichment analysis. Real-time polymerase chain reaction（Real-time PCR） was used to detect the mRNA expression of Gad，Slc32a1（VGAT） and brain-derived neurotrophic factor (BDNF） in nucleus accumbens. ResultCompared with the control group，the anxiety and depression-like behaviors in the model group increased，while the learning and memory ability decreased significantly. The content of neurotransmitter in serum decreased significantly. The content of pro-inflammatory factors increased significantly. The mRNA expression of Gad and Slc32a1（VGAT） in nucleus accumbens decreased significantly，while that of BDNF increased significantly. Compared with the model group，the anxiety and depression-like behavior of mice in verbascoside group was significantly relieved. The neurotransmitter content increased significantly，and the pro-inflammatory factors decreased significantly. The mRNA expression of Gad and Slc32a1（VGAT） in nucleus accumbens increased significantly，while that of BDNF decreased significantly.A total of 48 differentially expressed genes in nucleus accumbens and 43 differentially expressed genes in colon were screened by high-throughput sequencing. Differential genes in nucleus accumbens mainly focus on neuroactive ligand-receptor interaction， γ-aminobutyric acid（GABA）ergic synapse，synaptic vesicle cycle and other pathways. Colonic differential genes are mainly concentrated in GABAergic synapses，synaptic vesicle circulation，cyclic adenosine monophosphate（cAMP） signal pathway and other signal pathways. Compared with the control group，the mRNA expression of Gad and Slc32a1（VGAT） in nucleus accumbens of model group decreased significantly，while the mRNA expression of BDNF increased significantly. ConclusionVerbascoside has significant antidepressant effects. Its antidepressant effect may be related to the increase of monoamine neurotransmitters，the decrease of pro-inflammatory factors and the restoration of neurotransmitter homeostasis by increasing GABA，and it mainly acts through the signaling pathways such as neuroactive ligand-receptor interaction，GABAergic synapses，synaptic vesicle cycle and cAMP signaling pathway.