Objective:To establish a method for the content determination of triptolide in compound tripterygium wilfordil syrups by HPLC. Methods:The samples were analyzed on a Diamonsil C18 column(150 mm × 4. 6 mm,5μm), the mobile phase consisted of methanol and water (46∶54), the flow rate was 1. 0 ml·min-1, the detection wavelength was 224 nm, and the column temperature was kept at 30℃, and the injection volume was 10 μl. Results: The linear range of triptolide was 5. 66-33. 96 μg · ml-1 ( r =0. 999 8), and the average recovery was 99. 35% with RSD of 1. 02%(n=9). Conclusion:The method is simple and sensitive with good repeatability, and can be used to control the quality of triptolide in compound tripterygium wilfordil syrups.

Objective To investigate the localization and the morphological changes of manchette during mouse spermiogenesis.Methods Immunofluorescence staining with FITC and costaining with DAPI were used to demonstrate the cellular localization of the manchette at different stages during mouse spermiogenesis.The structural changes of the manchette were observed during the maturing of the spermatid.Results Immunofluorescence staining showed that manchette existed exactly around the nuclei of the spermatids.Manchette began to form,when the shape of the nucleus changed from spherical to slightly elongated.While the nucleus of the spermatids condensed and elongated at later stages,manchette moved gradually to the caudal position of the spermatids.At last,the manchette diminished as the spermatids became mature.During mouse spermiogenesis,manchette underwent a transition from a cap-like to a tubular configuration.ConclusionThe formation and diminishment of the manchette is in step with the condensation and elongation of the nucleus of the spermatid.Both the structural and positional changes of the manchette coincide with the changes of the nucleus.These results imply that manchette might play an important role in mouse spermiogenesis.

Objective To study the inhibition effect of non custodial terpenes-3β-alcohol to experimentally in-duced autoimmune encephalomyelitis in guinea pigs. Methods Different doses (25 mg/kg, 50 mg/kg and 100 mg/kg) of non custodial terpenes-3β-alcohol were given to the experimentally induced autoimmune encephalomyelitis model of guinea pigs by gavage for 8 weeks. Plasma levels of CD4+/CD8+, IL-1, IL-2, IL-6, IL-10, neuropeptide Y (NPY), beta endorphin (β-EP) , transforming growth factor-β(TGF-β), matrix metalloproteinase (MMP-2), nitric oxide synthase (NOS) and leuko-cyte differentiation antigen CD3 were assessed. The brain neuron morphology changes was observed under light microscopy while its ultrastructure changes was observed under electron microscope. NOS expression in neurons was observed through immunofluoresce technology. Results Non custodialterpenes-3β-alcohol inhibited the increase of plasma CD4+/CD8+, IL-1, IL-2, IL-6, IL-10, MMP-2, CD3 and NPY while decrease of plasmaβ-EP, brain TGF-β. It also increase NOS expres-sion in neuronal cytoplasm and maintained neuron morphology. Conclusion Non custodial terpenes-3β-alcohol inhibit-ed the experimental autoimmune encephalomyelitis in guinea pig.

Objective To investigate the expression and distribution of Diacylglycerol Kinase α (DGKα) mRNA in human hepatocellular carcinoma (HCC),and to explore the function of DGKα in the metastasis of hepatocellular carcinoma.Methods Tissues from 30 cases of HCC and 5 normal liver tissues were collected immediately after surgical resection.Semi-quantitative RT-PCR and in situ hybridization were used to detect the expression levels and distribution of DGKα mRNA,respectively.Results Semi-quantitative RT-PCR showed that the expression level of DGKα mRNA in HCC (0.798±0.317) and normal tissues (0.908±0.425) was significantly higher than those in carcinoma adjacent tissue with cirrhosis (0.205±0.102,P<0.05).In situ hybridization demonstrated that the number of DGKα mRNA positive hepatocytes in HCC [(57.6±6.3)/mm2] and normal tissues [69.8±8.7)/mm2] was significantly higher than those in carcinoma adjacent tissue with cirrhosis [(26.3±4.9)/mm2,P<0.05]; DGKα mRNA was expressed in the cytoplasm of hepatocytes in HCC and carcinoma adjacent tissue with cirrhosis,and in the nuclear of hepatocytes in normal tissues.Conclusion The present study suggests that DGKa may play important roles in carcinogenesis and progressing of HCC.

Objective:To express SARS-CoV nucleocapsid protein in Bac-to-Bac Baculovirus Expression System and analyze the antigenicity of the recombinant protein.Methods: The SARS-CoV nucleocapsid gene was amplified by PCR.The PCR product digested with BamHⅠand SalⅠrestriction endonucleases was cloned into vector pFastBac HTC of Bac-to-Bac Baculovirus expression system.Recombinant plasmid was transformed DH 10Bac cells to obtain the recombinant Bacmid DNA.Recombinant Bacmid DNA was transferred into Sf9 cells which were inducted to express the recombinant protein in High Five cells.After purified by Ni affinity chroma-tography ,the antigenicity of the recombinant protein was analyzed by Western blot and ELISA.Results:Recombinant plasmid was con-structed successfully.The recombinant protein with the relative molecular mass of 48 kD was efficiently expressed in High Five cells and purified successfully by Ni affinity chromatography.Western blot and ELISA analysis showed that the recombinant protein could be spe -cifically recognized by the monoclonal antibody to SARS-CoV N protein and immune serum from rabbits ,respectively.The recombinant protein can specifically reacted with serum from SARS patients ,not with serum from healthy persons and patients infected with hCoV-229 E and hCoV-OC43.Conclusion: SARS-CoV nucleocapsid protein has been expressed successfully in the Bac-to-Bac Baculovirus Expression System ,and obtained good antigenicity.It is preliminary deemed that it can't reacted with serum from patients infected with hCoV-229E and hCoV-OC43.

Objective To investigate the effect of creatine phosphate sodium on postoperative fatigue syndrome (POFS) in gastric carcinoma patients. Methods Nighty cases of radical resection in gastric cancer were randomly divided into control group (n=45) and treatment group (n=45).Normal saline (100 mL) and phosphocreatine sodium (2 g) was admin-istrated intravenously after anaesthesia induction in treatment group while only normal saline was given in control group . After operation, the two groups of patients were all treated with enteral and parenteral nutrition. General performance was ob-served in both groups. Fatigue score (VAS score) was measured on the day before the operation,the first,third and fifth day after operation. Body weight, triceps skinfold thickness (TSF), mid upper arm circumference (AMC), plasma total protein (TP), plasma albumin (ALB) and prealbumin (PA) were examined at the same time. Results The total effective rate of the control group and the study group were 73.3%and 95.6%respectively and the difference was of statistically significant. Fa-tigue scores (VAS score) was significantly difference in both groups pre- and post-operation time points, also between 2 groups. Body mass, TSF and AMC in time points postoperation were all lower than those in the time point of preoperation. TP, ALB and PA of two groups in postoperative time points were significantly different compared with those in preoperative time point. The above indexes were all decreased with postoperation time, and these indexes in treatment group were signifi-cantly better than those in control group. Conclusion Creatine phosphate sodium can alleviate postoperative fatigue, im-prove nutritious status and promote early rehabilitation of patients.

Objective To study the radiation-protective effect of diallyl disulfide(DADS)in germ cells of male mice. Methods Male mice were whole-body exposured to 4 Gy X-ray irradiation to establish a animal radiation damage model. Testicular histology, sperm motility and sperm deformity rate were observed, protein carbonyl content, malondialdehyde(MDA)and 8-hydroxy deoxyguanosine(8-OHdG)content were tested to assess the degrees of radiation damages of sperm cells and protective effect of DADS. Testicular tissue antioxidant enzymes such as superoxide dismutase (SOD),catalase(CAT),glutathione peroxidase(GSH-px)activity and glutathione(GSH)content were examined. The expression of Nrf2 signal protein was detected to explore the radiation-protective mechanism of DADS. Results Compared with the pure exposure group, the testicular tissue damages in the DADS pretreatment group were milder, sperm motility increased significantly(P< 0.05)and sperm deformity rate decreased(P< 0.05), and the protein carbonyl content, MDA and 8-OHdG levels significantly reduced(P< 0.05). The antioxidant indexes of SOD, CAT, GSH-px and GSH were markedly improved(P< 0.05),and the expression of Nrf2 was dramatically enhanced. Conclusions DADS has a protective effect on acute radiation injury in germ cells of male mice by means of anti-oxidative ability.

BACKGROUNDThere have been a lot of studies about surface adhesion molecule (CD44) in recent years, however study on osteopontin (OPN) is still few. The aim of this study is to investigate the levels of OPN and CD44v6 in lung cancer and their correlation.

METHODSOPN and CD44v6 expression were detected in 78 lung cancer tissues by immunohistochemistry.

RESULTSThe positive rate of OPN and CD44v6 expression in non-small cell lung cancer (NSCLC) was 58.46% and 66.15% respectively, but no positive case in small cell lung cancer. The expression of OPN and CD44v6 in NSCLC was closely related to TNM stages (P < 0.01), but not to cell differentiation (P > 0.05). There was a remarkably positive correlation between the expression of OPN and CD44v6 (r=0.255, P < 0.05).

CONCLUSIONSThe co-expression of OPN and CD44v6 may be involved in progression and metastasis of lung cancer. The interrelationship of OPN and CD44v6 was coordinative with development and metastasis of lung cancer. It might be helpful to predict the metastasis and prognosis of NSCLC.

Adult ; Humans ; Male ; Mediastinal Cyst

OBJECTIVETo investigate the characteristics and dynamic changes of serum neutralizing antibody response in patients with primary infection of dengue virus type 1 (DENV-1).

METHODSSerum samples were obtained from the same patients with primary infection of DENV-1 within 2 weeks after symptom onset in 2006 and in 2010. A group-specific DENV NS1 capture ELISA-based micro-neutralizing test (ELISA-MNT) capable of detecting neutralizing antibodies against all the 4 serotypes of DENV was used to test the neutralizing antibody titers against DENV in the serum samples. The neutralizing antibody titers against a standard strain and 2 clinically isolated strains of DENV-1 were detected in serum samples collected in 2010.

RESULTSCross-reactive neutralizing antibody response against all the 4 serotypes of DENV was found in both of the serum samples collected in 2006 and 2010, but the samples collected in 2006 showed stronger cross-reactive neutralizing antibody responses. The neutralizing antibody against DENV-2, rather than the anticipated DENV-1 antibody, had the highest titer in the samples collected in 2006, whereas the antibody against homologous DENV-1 had the highest titer in the samples obtained in 2010. The neutralizing antibody titers against the homologous DENV-1 was significantly higher in samples collected in 2010 (U=86.500, P=0.000), which also demonstrated significantly different neutralizing antibody titers against the 3 different strains of DENV-1 (Χ(2)=12.123, P=0.002).

CONCLUSIONThe production of cross-reactive neutralizing antibodies between the 4 serotypes of DENV is a characteristic of DENV infection, particularly during early infection, but only the homologous neutralizing antibody increases obviously over time. The titers of the neutralizing antibodies against different strains, even of the same serotype, may differ distinctly.

Antibodies, Neutralizing ; blood ; Antibodies, Viral ; blood ; Cross Reactions ; Dengue ; blood ; immunology ; Dengue Virus ; classification ; immunology ; Humans ; Neutralization Tests