Pathogenesis and Differentiated Treatment Strategies of Childhood Tic Disorders Based on WANG Xugao's “Thirty Methods of Treating the Liver” / 中医杂志

Based on WANG Xugao's “thirty methods of treating the liver”， it is believed that the occurrence and development of childhood tic disorders follow the dynamic progression from liver qi disease to liver fire disease and then liver wind disease. The basic pathogenesis of three stages are characterized by binding constraint of liver qi， liver fire hyperactivity， and internal stirring of liver wind. Moreover， liver-blood deficiency and stagnation， and malnutrition of liver yin as the main point in terms of the imbalance of liver qi， blood， yin， and yang should be considered， as well as the imbalance relationship of the five zang organs such as the involvement of other organs and the gradually reach of the other organs. Guided by the principles of “thirty methods of treating the liver”， the treatment of tic disorders in liver qi stage should focus on soothing the liver and rectifying qi， soothing the liver and unblocking the collaterals， using Xiaochaihu Decoction （小柴胡汤） and Sini Powder （四逆散）. The treatment of tic disorders in liver fire stage involves clearing， draining and resolving liver heat， using Longdan Xiegan Decoction （龙胆泻肝汤）， Xieqing Pill （泻青丸）， Danggui Longhui Pill （当归龙荟丸）， and Huagan Decoction （化肝煎）. The treatment of tic disorders in liver wind stage involves extinguishing wind and subduing yang， using Lingjiao Gouteng Decoction （羚角钩藤汤） and Liuwei Dihuang Pill （六味地黄丸）. Throughout the treatment process， attention should be paid to harmonizing the liver's qi， blood， yin， and yang， as well as addressing the pathology of other organs.

Rupture of hepatic artery pseudoaneurysm after liver transplantation: a report of three patients / 中华肝胆外科杂志

Objective:To report on 3 patients who presented with rupture of hepatic artery pseudoaneurysm after liver transplantation.Methods:From April 2010 to April 2019, 3 patients with hepatic artery pseudoaneurysm rupture after liver transplantation treated at the Department of Hepatobiliary and Pancreatic Surgery, Henan Provincial People's Hospital were studied. The possible causes, clinical manifestations, diagnosis and treatment were retrospectively analyzed.Results:Rupture of hepatic artery pseudoaneurysm occurred on the19th, 28th and 63th days after transplantation. The 3 patients all presented with hematochezia and abdominal pain, while 2 patients presented with hematemesis. Two patients had bile leakage and abdominal infection. All the 3 patients presented with fever. Patient 1 who was diagnosed by laparotomy died of liver failure. Patient 2 underwent interventional embolization of hepatic artery and died of liver failure also. Patient 3 underwent surgical resection of the pseudoaneurysm followed by hepatic artery reconstruction, but died of repeat abdominal hemorrhage.Conclusion:Hepatic artery pseudoaneurysm after liver transplantation has a long latent period and is difficult to diagnose at an early stage. Early detection of this life-threatening complication is the key to improve survival. Early treatment of biliary leakage, abdominal infection and other complications help to prevent development of pseudoaneurysms.

Expression of interleukin-33 and vascular endothelial growth factor-C in gastric cancer and its clinical significance / 重庆医学

Objective To investigate the expression of interleukin-33(IL-33)and vascular endothelial growth factor C(VEGF-C)in gastric cancer tissues and serum,and to explore the relationship between these two indicators and gastric cancer lymph node metastasis.Methods The levels of IL-33 and VEGF-C in the tissues of gastric mucosa and serum were detected by immunohistochemical SP method and enzyme-linked immunosorbent assay(ELISA)in 98 patients with gastric cancer and 36 healthy subjects.Results The expression rates of IL-33 and VEGF-C in gastric cancer were 67.35％and 74.49％,which were significantly higher than the rates in normal gastric tissue(47.22％and 61.11％).The difference was statistically significant(P<0.01).The expression of IL-33 and VEGF-C was correlated with the degree of tumor differentiation,tissue infiltration,lymph node metastasis,distant metastasis and clinical stage(P<0.05).The positive rates of IL-33 and VEGF-C in gastric cancer lymph node metastasis group were higher than those in non-lymph node metastasis group(P<0.05).The serum concentrations of IL-33 and VEGF-C in patients with gastric cancer were(50.24±13.08)pg/mL and(210.73±58.35)pg/mL,respectively,which were higher than those in healthy control group(P<0.05);the expressions of serum concentration of IL-33 and VEGF-C in the cases with lymph node metastasis were higher than those without lymph node metastasis and the difference was statistically significant(P<0.05).Conclusion High levels of IL-33 in gastric carcinoma patients might induce the secretion of VEGF-C,promote lymph node metastasis,and be applied as an important index of the appraisal to the prognosis of gastric cancer.

Optimization of triple plasmids transfection into HEK293 cells mediated by polyethylenimine / 生物医学工程学杂志

In the present study, packaging system composed of pAAV-CMV-GFP, pAAV-RC and pHelper were transfected into human embryonic kidney 293 cells (HEK293 cells) mediated by polyethyleneimine (PEI) to explore an optimal transfection condition. Different total plasmid DNA dosages (1, 2, 3, 4, 5, 6 μg) and different PEI/Plasmid ratios (1:1, 3:1, 5:1, 7:1) were tested with detection of green fluorescence protein (GFP) with ImagePro Plus6. 0 Software. Then transfection efficiency of the optimized transfection system was further observed for different time periods(12, 24, 36, 48, 60, 72 h). The results showed that total plasmid dosage of 4 μg/well with PEI/plasmid ratio of 3 : 1-5 : 1 was an efficient transfection condition. Transfection efficiency-time curve was an S-shaped curve. Transfection efficiency reached a plateau at 60 h after transfection. The optimized conditions for PEI-mediated transfection at the optimal time result in enhanced transfection efficiency of triple plasmid into HEK293 cells.

A novel packaging system of recombinant AAV5/5 vector / 生物工程学报

We developed a scalable AAV5/5 vector packaging system by using replication competent recombinant herpes simplex type 1 virus as helper virus. The fragment containing rep and cap genes of AAV5 was inserted into the non-necessary gene (UL2) of HSV1 genome, resulting in the helper virus rHSV1-rep5cap5. An AAV5/5 vector pAAV5neo carrying two AAV5 ITRs was constructed by inserting a neo gene expression cassette into the plasmid backbone of pAV5CMV-GFP. pAAV5neo-EGFP was constructed by inserting EGFP gene into pAAV5neo. BHK21 cell was transfected with pAAV5neo-EGFP and cultured in the presence of G418. EGFP expression positive monoclonal cells were picked up, and one that produced rAAV5/5-EGFP with the highest efficiency under the help of rHSV1-rep5cap5 was chosen as the production cell line named as C020. rAAV5/5-EGFP was produced by infecting C020 cells with rHSV1-rep5cap5, and crudely purified by our previous method of 'chloroform treatment-PEG8000/NaCl precipitation- chloroform extract'. rAAV5/5-EGFP preparation with high purity was obtained by ultrafiltration with molecular weight cut-off value of 100 kDa. SDS-PAGE stained with Coomassie brilliant blue R250 showed clearly specific pattern of three bands of AAV capsid proteins. rAAV5/5-EGFP was also assayed using negative stain transmission electron microscopy and the majority of the virus particles were found solid. About 30% green fluorescent cells could be seen after infecting HEK293 cells with rAAV5/5-EGFP 24 h at 1 x 10(5) vg/cell. In conclusion, we have established an efficient AAV5/5 vector production system and could produce recombinant AAV5/5 virus in large amounts for gene therapy research.

Inhibition of Luciferase expression in mammalian cells by AAV vector plasmid mediated Luciferase shRNA / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>Constructing a plasmid containing the shRNA of luciferase to suppress the expression of luciferase in BHK-21 cell.</p><p><b>METHODS</b>A 334 bp human U6 snRNA promoter was amplified from human genomic DNA by PCR and ligated to a 21 bp reverse repeated motif of luciferase target sequence with 9 bp spacer and AAV plasmid pSNAV. The recombinant pSNAV/U6/Luc plasmid cotransfected with pMAMneoLuc or transfected luciferase cell line to detect the effect of luciferase expression separately.</p><p><b>RESULTS</b>pSNAV/U6/luc suppresses the luciferase expression from pMAMneoLuc by 50% and luciferase cell line by 70%.</p><p><b>CONCLUSIONS</b>The results showed that the short hairpin RNA of luciferase can efficiently suppress its expression in BHK-21.</p>