Acupuncture Points ; Acupuncture Therapy ; methods ; Adult ; Aged ; Catgut ; Female ; Humans ; Male ; Middle Aged ; Phlebotomy ; Trigeminal Neuralgia ; therapy

Objective To obtain the fusion genes of several human orphan G protein coupled receptors (oGPCRs) with Gi1α subtype of G protein and their expression system. Methods The whole open reading frames of GPR45, GPR85, GPR174 and Gilα were cloned by RT-PCR from HepG2 cDNA separately,and the corresponding fusion genes were amplified by overlap extension PCR. Then, the fusion genes-containing pBacmids were successfully constructed with the Bac-to-Bac baculovirus expression system indicated by specific transposition and virus recombination. The insect Sf9 cells were transfected with pBacmid-oGPCRs-Gi1α, and the supernatant containing recombinant virus was harvested. With the supernatant, insect Sf9 cells were infected under an optimized condition (MOI=5, infection time=72 h) and the fusion proteins were prepared and detected by Western blotting.Results The three fusion genes of GPCR45, GPR85 or GPR174 with Gi1α were obtained. The corresponding fusion proteins could be properly prepared in Sf9 cells.Conclusion Human oGPCRs could be fused with Gilα, and the fusion genes could be expressed using the Bac-to-Bac baculovirus expression system in insect Sf9 cells.

Objective To explore the efficacy of endoscopic sphincterotomy with small incision combined with balloon dilatation (sEST+EPBD)in the treatment of patients with choledocholithiasis and juxtapapillary duodenal diverticula (JPDD).Methods From January 2011 to January 2015 ,149 patients with choledocholithiasis and JPDD who underwent endoscopic retrograde cholangio-pancreatography (ERCP)were enrolled.Among them,60 patients were in sEST+EPBD group and 89 were in endoscopic sphincterotomy (EST)group.Success rate of ERCP and first-time stone removal,changes of total bilirubin (TBil)and direct bilirubin (DBil)levels,as well as the incidence of postoperative complications between the two groups were compared.Chi-square test or t-test was performed for statistical analysis. Results The ERCP success rate sEST+EPBD group was 100.0% (60/60),and the first-time success rate of stone removal was 91 .7%(55/60);correspondingly,ERCP success rate of EST group was 98.9%(88/89),and the success rate of first-time stone removal was 77.5 %(69/89).There was no statistically significant difference in success rate of ERCP between the two groups (χ2 =0.19,P =0.410).The first-time success rate of stone removal of sEST +EPBD group was higher than that of EST group,and the difference was statistically significant (χ2 =5 .53,P =0.020).After operation,the TBil level of sEST+
EPBD group was (152.62 ±109.04 )μmol/L,which was lower than that before operation ((266.02 ± 143.31)μmol/L),and the difference was statistically significant (t =4.88,P <0.01 ).After operation, the DBil level of sEST +EPBD group was (87.13 ±65 .90)μmol/L,which was lower than that before operation ((175 .70 ± 100.53 )μmol/L),and the difference was statistically significant (t = 5 .71 ,P <0.01).After operation,the TBil level of EST group was (251 .90 ±247.90)μmol/L,which was lower than that before operation ((340.20 ±176.20 )μmol/L),and the difference was statistically significant (t=2.74,P <0.05).After operation,the DBil level of EST group was (168.10±140.60)μmol/L,which was lower than that before operation ((228.40 ±139.60 )μmol/L),and the difference was statistically significant (t = 2.87,P = 0.005).The complication rate of sEST + EPBD group after operation was 8.3%(5/60),which was lower than that of EST group (20.2%,18/89 ),and the difference was statistically significant (χ2 =3.88,P =0.049 ).Conclusion sEST+EPBD could increase the first-time success rate of stone removal in patients with choledocholithiasis and JPDD,and it is a safe and effective treatment.

Adult ; Aged ; Biomarkers, Tumor ; blood ; Carcinoma, Hepatocellular ; diagnosis ; enzymology ; Female ; Humans ; Liver Neoplasms ; diagnosis ; enzymology ; Male ; Middle Aged ; RNA, Messenger ; blood ; classification ; alpha-Fetoproteins ; analysis ; gamma-Glutamyltransferase ; blood ; classification

To evaluate the efficacy of Tuina and Chinese patent drug Shuxuetong injection in preventing patients undergoing total knee arthroplasty from deep venous thrombosis and in functional rehabilitation.

Objective To study the prevention and cure effect of immunoregulation therapy on postoperative infectious complications of calculous obstructive jaundice.Methods Prospective,randomized,blind and controlled clinical analysis of 40 patients conforming to the standard of calculous obstructive jaundice was carried out.The patients were divided into two groups at random.One was control group (n=20) with regular therapy,and the treatment group (n=20) with ulinastatin plus thymosin-α1 on the base of regular therapy for 1 week.The immunological indexes were determined before and after therapy on the 1st,3rd,5th、7th and 14th day,including the changes in lymphocyte count,CD14+ monocytes human leukocyte antigen (locus) DR (HLA-DR),and the incidence of postoperative infection were observed.Results The incidence of postoperative infectious complications in treatment group (10％) was remarkable lower than that in control group (30％,P＜0.01).There was significant difference between two groups (P＜0.05).After the 3rd,up to 14th day of therapy,the counts of lymphocyte and CD14+ monocytes HLA-DR were significantly higher than those in control group (all P＜0.05).Conclusions Immunoregulation therapy can improve the prognosis of obstructive jaundice patients in a period of 14 days of observation,and lymphocyte counts and CD14+monocytes HLA-DR were increased significantly,showing that immuno suppression can be ameliorated.Immunoregulation therapy could effectively prevent infectious complications of calculous obstructive jaundice.

Objective To investigate the effect of herpes simplex virus thymidine kinase/ganciclovir (HSV-TK/GGV)system driven by human alpha fetoprotein(AFP)enhancer on hepatocellular carcinoma(HCC)cells in vitro and in vivo.Methods HCC-specific eukarotypic expression vector carrying suicide gene driven by AFP enhancer(pAFP-cDNA3.1-TK)was constructed.The plasmid was trasfected to AFP-positive HepG2 cells and AFP-negative SMMC7721 cells by liposomes.Protein and mRNA expressions of TK were detected by RT-PCR or Western blot.The survival rates of HCC cells were detected by methyl thiazolyl tetrazolium assay.The effects of GGV on the in vitro proliferation,survival and apoptosis of HCC cells were observed,and the inhibitive effect of GGV on the survival of HCC cells in vivo was also detected.All data were analyzed by using the t test.Results The pAFP-cDNA3.1-TK was successfully constructed and transfected to the HCC cells.The protein and mRNA expressions of TK were detected in AFP-positive HepG2 cells.GGV dose-and time-dependently inhibited the growth and induced the apoptosis of HepG2 cells in vitro,but it had no effect on SMMC7721 cells.No protein or mRNA expression of TK was detected in the SMMC7721 cells.There was a significant difference on the inhibitory effects of GGV on HepG2 cells and SMMC7721 cells(t =2.58,2.73,3.12,P ＜0.05).GGV specifically inhibited the growth of AFP-positive HepG2 cells,and the inhibition rate was 46%;the growth of AFP-negative SMMC7721 cells was not influenced by GGV.There was a significant difference in the inhibitive effect of GGV on the growth of HepG2 cells and SMMC7721 cells(t = 3.36,P ＜ 0.05).Conclusion HSV-TK/GGV systemdriven by human APF enhancer kills APF-positive HCC cells and inhibits the growth of HCC cells.

The major antigen region of E2 gene of Hog Cholera Prevalent Strain (Guangxi Yuling Strain) and Chinese Hog Cholera Lapinised Virus (C-strain) derived from hog and rabbit spleen tissue, was amplified by reverse transcription polymerase chain reaction(RT-PCR) and the nested Polymerase Chain Reaction (nPCR). After the amplified fragments were cloned into the expression vector pPROEX-HTb, the recombinant plasmids pPROEX-GXYL and pPROEX-C were obtained. The insert position, the size and the reading frame were right by PCR, restriction digestion and the sequence analysis. SDS-PAGE indicated that both of the reciepient germs transducted and induced by the recombinant plasmids pPROEX-GXYL and pPROEX-C could express the major antigen region of E2 gene. Western-blot indicated that the expressed antigen protein could be recognized by the positive serum of CSFV.
Blotting, Western ; Cloning, Molecular ; Escherichia coli ; genetics ; Polymerase Chain Reaction ; Recombinant Proteins ; biosynthesis ; Viral Envelope Proteins ; biosynthesis ; genetics ; immunology

E2 gene of classical swine fever virus (CSFV) was cloned into secretory pPIC9K Pichia pastoris expression vector. After being linearized by digestion, the vector was transformed into Pichia pastoris by electroporation to integrate with the genome, the transformants with high copies were screened by G418 and were induced to express with methonal. The results of SDS-PAGE and Western blot demonstrated that the supernatant of the induced P. pastoris culture contained protein E2. The results of the study on the immunological activity indicated that the protein E2 expressed in P. pastoris can elicit animal bodies to produce antibodies against protein E2.
Animals ; Antibodies, Viral ; immunology ; Antigens, Viral ; genetics ; immunology ; Classical swine fever virus ; genetics ; immunology ; Cloning, Molecular ; Gene Expression ; Pichia ; Rabbits ; Swine ; Viral Envelope Proteins ; genetics ; immunology

OBJECTIVETo observe the effect of Danshen Chuanxiongqin Injection (DCI) on the myocardial damage of unstable angina (UA) patients undergoing percutaneous coronary intervention (PCI).

METHODS76 UA patients were randomly assigned to the test group (39 cases) and the control group (37 cases). Routine treatment of Western medicine and PCI were given to all patients. But 10 mL DCI was intravenously dripped to patients in the test group by adding in 250 mL normal saline three to five days before PCI, once daily, lasting for 7 to 10 days. Changes of platelet P-selectin positive expressions before and after medication, the thrombolysis in myocardial infarction (TIMI), myocardial perfusion (TMP) score, levels of creatine kinase-myocardial band (CK-MB) and troponin I (TnI) after PCI were observed in the two groups.

RESULTSAfter medication the P-selectin positive expressions was lower in the test group than in the control group (2.45% +/- 1.42% vs 4.43% +/- 1.79%, P<0.05). After PCI the incidence of TMP 0/1, the levels of CK-MB and TnI were lower in the test group than in the control group at the same phase, showing statistical difference (P<0.05). No cardiac event occurred in the two groups during the hospital stay.

CONCLUSIONAdministration of DCI before PCI could effectively inhibit the activation of platelets, improve post-operative myocardial blood perfusion, and lower the incidence of the myocardial damage.

Adult ; Aged ; Aged, 80 and over ; Angina, Unstable ; blood ; therapy ; Creatine Kinase ; blood ; Drugs, Chinese Herbal ; pharmacology ; Female ; Humans ; Male ; Middle Aged ; Myocardium ; metabolism ; pathology ; Percutaneous Coronary Intervention ; Phenanthrolines ; pharmacology ; Pyrazines ; pharmacology ; Troponin I ; blood