In order to construct a prokaryotic expression vector of human receptor syt II N-fragment and to express recombinant MBP-Syt fusion protein in E.coli and to purify and identify its activity. According to codon preference of E.coli, a DNA fragment encoding human syt II N-fragment was synthesized, and then cloned into prokaryotic vector pMAL-c2x for sequencing. Then the recombinant plasmid pMAL-Syt was introduced into E.coli ER2566 by transformation for expression and the obtained engineered bacteria were induced by IPTG. The fusion protein was purified by amylose resin affinity chromatography and identified by SDS-PAGE and Western blot. The binding activity of the protein was determined by ELISA. It is concluded that MBP-Syt protein is of good binding activity.

Objective:To express the B subunit of Shiga-like toxin type Ⅱ,and analyze its expression form and receptor-binding activity.Methods:The slt2b gene was obtained from EHEC O157∶H7 by PCR,and cloned to the expression vector pET22b(+).The genetically engineered bacteria pET22b(+)-stx2B/BL21 expressed the recombinant StxB after induced with IPTG.The renatured inclusion bodies were purified by ion exchange chromatography.The expression form of rStx2B was investigated by denaturing and native electrophoresis.The receptor-binding activity was confirmed by fluorescence detection and flow cytometer.Result:The constructed genetically engineered bacteria expressed the rStx2B at a high level.The purified protein was obtained after denaturation,renaturation and ion exchange chromatography.According to the denaturing and native electrophoresis,the rStx2B was expressed in a dimmer form,which consists of two monomers cross linked with disulfide bridge.The rStx2B showed good receptor-binding activity by Hela-binding assay.Conclusion:The genetically engineered bacteria were constructed successfully.The receptor-binding activity of rStx2B was independent of the pentamers.

This paper introduces the principles of the medical gas central supply system and its structure, Classification, standards, flow chart and its advantages, and its quantity guarantee and quality control.
Equipment Design ; Equipment Safety ; Equipment and Supplies, Hospital ; standards ; Gases ; Humans ; Quality Control

OBJECTIVETo explore the effect of emodin on endoplasmic reticulum (ER) stress of pancreatic acinar AR42J cells.

METHODRat pancreatic acinar AR42J cells were cultured in 6-well plates, and divided into the normal control group, the model group (with the final concentration at 1 x 10(-7) mol · L(-1) for cerulean and lipopolysaccharide at 10 mg · L(-1)) and the emodin group (10, 20, 40 μmol · L(-1)). Cells in each group were cultured in three multiple pores for 24 h, and their supernate was removed after cell attachment. The normal control group was added with haploids, the model group was added with the modeling liquid for haploids, and the treatment groups were added with different concentrations of emodin at 15-20 min before the modeling liquid. The cells were continuously cultured for 3 h under 37 °C and 5% CO2. Their intracellular protease and lipase expressions were detected with kits. The cellular morphology was observed under optical microscope. The level of calcium in endoplasmic reticulum was measured under laser confocal microscopy. Western blot assay were used to determine the protein expression of ER-related signaling molecules.

RESULTEmodin could significantly inhibit levels of amylase, lipase and intracellular calcium and ER.

CONCLUSIONEmodin could reduce pancreatic acinar cell injury induced by the combination of cerulean and lipopolysaccharide. Its action mechanism is correlated with the inhibition of intracellular calcium overload and ER stress.

Animals ; Calcium ; metabolism ; Cell Line, Tumor ; Emodin ; pharmacology ; Endoplasmic Reticulum Stress ; drug effects ; Pancreatic Neoplasms ; metabolism ; pathology ; Rats ; Unfolded Protein Response ; drug effects

To produce antibodies capable of neutralizing botulinum neurotoxin type B(BoNT/B),We cloned the carboxy-terminal end of Hc containing the major determinants responsible for specific toxin,induced and purifed.The heavy-chain and kappa light-chain variable region gene repertoire of immunoglobulin were amplified individually from the spleen cell mRNA by RT-PCR and joined as a single-chain Fv(scFv)DNA fragment.These fragment were cloned into the phagemid pCANTAB5E and the phage display library was constructed.Results showed that the high affinity scFv was obtained after 4 rounds of panning,with its DNA sequence conforming to that of mouse antibody.

OBJECTIVETo investigate the bonding properties and interface characterization of a domestic 3mol yttrium-stabilized tetragonal zirconium polycrystal (3Y-TZP) framework fired on with 4 different veneering ceramics for zirconia.

METHODS4 different commercial veneering ceramics for zirconia (VITA VM9, SHOFU VINTAGE ZR, IPS e.max Ceram, Cercon ceram kiss) were sintered on 3Y-TZP rectangulars (15 mmx5 mmx5 mm) according to the manufacturers' instructions for shear bond strength test, a metal-ceramic system(Ni-Cr alloy/VITA VMK95) was fabricated in the same type as a control group. Two bilayered specimens (3Y-TZP/VITA VM9, Ni-Cr/VMK95) were prepared for scanning electron microscope (SEM) and energy distribution spectrum (EDS).

RESULTSThe values of shear bond strength test were (18.83 +/- 1.77) MPa for 3Y-TZP/VITA VM9, (23.83 +/- 7.05) MPa for 3Y-TZP/SHOFU VINTAGE ZR, (17.87 +/- 2.30) MPa for 3Y-TZP/IPS e.max Ceram, (22.26 +/- 7.45) MPa for 3Y-TZP/Cercon ceram kiss, (20.55 +/- 5.13) MPa for Ni-Cr alloy/VITA VMK95. There was no statistically significant between all-ceramic groups and the control group (P > 0.05). The failure modes in all-ceramic groups showed predominately adhesive at the interface. SEM showed the 3Y-TZP/VITA VM9 contacted tightly at the interface, while EDS detected Si element diffused into 3Y-TZP material.

CONCLUSIONThe results indicate that domestic 3Y-TZP has a good interface compatibility with 4 commercial veneering ceramics, as a dental framework material, it can satisfy the clinical requirements.

Ceramics ; Dental Bonding ; Dental Porcelain ; Dental Stress Analysis ; Dental Veneers ; Materials Testing ; Yttrium ; Zirconium

OBJECTIVETo research the treatment effect of complex prescription of Chinese crude drug in BALB/c athymic mice with human liver cancer, which were built by Bel-7402.

METHOD48 male BALB/c athymic mouse models were built by Bel-7402 with an indirect method. After 24 hours of postoperation, the 48 athymic mice were distributed randomly into 4 groups which were treated by intragastric administration with complex prescription of Chinese crude drug that had been deliquated into 3 groups by the different density as the low, middle, and high and FT207 (Tegafur) for 4 weeks. At last, athymic mice were put to death and PTEN was detected in hepatic tissue, latero-cancer tissue and cancer tissue by immunohistochemistry (PowerVision Two-Step Histostaining Reagent).

RESULTAll of the 48 athymic mice survived 12 to 28 days (Ms 24 days) and every mouse with liver cancer demonstrated by dissection. The result of immunohistochemistry represents that the intensity of PTEN in latero-cancer tissue is the highest, and then the hepatic tissue, the lowest is cancer tissue, P < 0.01. It also represents that the intensity of PTEN in treatment groups (A, B, C) is more higher than the control group (D), P < 0.05 or P < 0.01, and group B is the highest in the treatment groups, P < 0.05 or P < 0.01. However, there is no significant statistic difference between group A and group C.

CONCLUSIONThe higher expression of PTEN in the laterocancer tissue can represent the protective reaction of stress of the organism. And anticancer effect of this complex prescription of Chinese crude drug relates to an eligible density of it. Mechanisms of this complex prescription of Chinese crude drug healing HCC may partially be explained by enhancing the expression of PTEN in liver.

Animals ; Antineoplastic Agents, Phytogenic ; isolation & purification ; pharmacology ; Carcinoma, Hepatocellular ; drug therapy ; enzymology ; pathology ; Cell Line, Tumor ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Humans ; Immunohistochemistry ; Liver ; drug effects ; enzymology ; pathology ; Liver Neoplasms ; drug therapy ; enzymology ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; PTEN Phosphohydrolase ; metabolism ; Phytotherapy ; Plants, Medicinal ; chemistry ; Random Allocation ; Xenograft Model Antitumor Assays

OBJECTIVETo explore the efficacy and safety of acupuncture for post-stroke depression (PSD).

METHODSThe randomized, double-blind control study was designed. 43 post-stroke patients with current major depression episode (DSM-IV) were randomly assigned to an observation group (23 cases) and a control group (20 cases). The conventional symptomatic, supportive and anti-infection treatment in neurological internal medicine was applied to all of the cases in two groups. In observation group, acupuncture was applied to Baihui (GV 20), Yintang (EX-HN 3), Sishencong (EX-HN 1), Taichong (LR 3), etc. Additionally, the placebo was taken orally. In control group, acupuncture was applied to non-acupoint spots (5 mm lateral to the acupoints selected in observation group) with shallow needling technique. Moreover, Fluoxetine was taken orally. The treatment lasted for 6 weeks. Hamilton Depression Scale (HAMD), Asberg antidepressant side effect scale (ASES), adverse response of acupuncture and efficacy were compared between two groups.

RESULTSThe total effective rate was 73.9% (17/23) in observation group and was 80.0% (16/20) in control group, indicating equivalent efficacy between two groups. After treatment, HAMD score was reduced remarkably as compared with that before treatment in two groups (P < 0.05). ASES scores in 4 weeks of treatment and after treatment in observation group and ASES score after treatment in control group were reduced remarkably as compared with those before treatment (all P < 0.05). ASES scores in 4 weeks of treatment and after treatment in observation group were reduced much more remarkably as compared with those in control group (both P < 0.05). The incidences of adverse response of acupuncture were 13.0% (3/23) and 15.0% (3/20) respectively in observation group and control group, indicating that the adverse response was transient and had not recurred after symptomatic measures.

CONCLUSIONAcupuncture for PSD is as effective as fluoxetine, without obvious drug-induced adverse reaction involved.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Depression ; etiology ; therapy ; Female ; Humans ; Male ; Middle Aged ; Stroke ; complications ; Treatment Outcome

OBJECTIVETo investigate the reno protective effect of Shenhua recipe on the experimental model of 5/6 renal ablation.

METHOD5/6 renal ablation rats were underlying this experiment. They were administered Shenhua, irbesartan respectively by gavage during 12 weeks. Body weight, systolic blood pressure, proteinuria, Scr, BUN, total protein, albumin, Glycero and cholesterol were measured. Histologic glomenular and tubulointerstitial damage scores were measured at 12 weeks.

RESULTThe treated groups showed significantly less histologic glomerular and tubulointerstitial damage scores at 12 weeks. The plasma albumin were higher ( P < 0.05), urine protein excretion rates, serum cholesterol and creatinine were lower than in nontreated group, but arterial blood pressure was not significantly different in the three Shenhua treated groups compared with nontreated group.

CONCLUSIONShenhua can retard the progression of chronic renal injury in the 5/6 renal ablation without changes in systolic blood pressure.

Albumins ; metabolism ; Animals ; Astragalus membranaceus ; chemistry ; Atractylodes ; chemistry ; Blood Pressure ; drug effects ; Cholesterol ; blood ; Creatinine ; blood ; Curcuma ; chemistry ; Disease Progression ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Kidney Failure, Chronic ; etiology ; pathology ; prevention & control ; Kidney Glomerulus ; pathology ; Male ; Nephrectomy ; adverse effects ; methods ; Plants, Medicinal ; chemistry ; Rats ; Rats, Wistar

Objective To evaluate the clinical efficacy and safety of Shenfu injection in the treatment of leukopenia induced by chemotherapy docetaxel, epirubicin and cyclophosphamide adjuvant chemotherapy ( TEC chemotherapy) after surgery of breast cancer.Methods A total of 106 patients with breast cancer were randomly divided into control group ( n=53 ) and treatment group ( n =53 ) .Control group was re-ceived 75 mg? m-2 docetaxel, intravenous infusion on day 1 +50 mg? m-2 epirubicin, intravenous infusion on day 1 +500 mg? m-2 cy-clophosphamide, intravenous infusion on day 1, 3 weeks for a course. Treatment group was given 50 mL Shenfu injection added 500 mL of 5%glucose intravenous infusion and plus the treatment of control group, 1 to 10 days for a course.Patients of two groups were received 2 courses of treatment.The clinical efficacy, the incidence of leukopenia, white blood cell count to improve the rate and quality of life before and after treatment, and adverse drug reactions in two groups were compared.Results After treatment, total effective rate in treatment group was significantly higher than that in control group (84.91%vs 69.81%, P<0.05).After treatment 7,10 d, leukopenia rates in treatment group were 9.43%and 11.32%, significantly lower than 37.74%and 30.19%in control group (P<0.05).After treatment, the white blood cell count in the two groups all decreased than those before treatment, and the degree of decline in treatment group was higher than that in control group (P<0.05).After treatment, the rate of improvement of quality of life in treatment group was significantly higher than that in control group (88.68%vs 54.72%, P<0.05).The incidence of adverse drug reactions in treatment group was significantly lower than that in control group ( P<0.05).Conclusion Shenfu injection has a definitive clinical efficacy for the treatment of chemotherapy-induced leukopenia, which may improve immune function and the quality of life, worthy of further promotion in clinical practice.