OBJECTIVETo explore the effect of Qiling Decoction (QD) combined highly active antiretroviral treatment (HAART) on expression levels of peripheral blood Th17 and Treg cells in HIV/AIDS patients.

METHODSTotally 55 HIV/AIDS patients were randomly assigned to the treatment group (28 cases) and the combination group (27 cases). Besides, 21 HIV negative patients were recruited as the healthy control group. Those in the treatment group received HARRT alone, while those in the combination group received HAART combined QD. The observation lasted for 24 weeks. Meanwhile, according to peripheral blood CD4+ T cell counts before treatment, HIV/AIDS patients were assigned to three subgroups. For patients in subgroup 1, 1 cells/microL < CD4+ T cell counts < or = 100 cells/microL; For patients in subgroup 2, 101 cells/microL < CD4+ T cell counts < or = 200 cells/lL; For patients in subgroup 3, 201 cells/microL < CD4+ T cell counts < or = 350 cells/microL. Expression of peripheral blood Th17 and Treg cells, and number of CD4+ T cell counts were detected using flow cytometry (FCM)in HIV/AIDS patients at the pre-treatment baseline, week 4, 12, and 24, as well as those in the healthy control group.

RESULTSCompared with the healthy control group, CD4+ T cell counts and the baseline expression level of Th17 cells in the peripheral blood of HIV/AIDS patients significantly decreased, the expression level of Treg cells significantly increased P < 0.01). Compared with before treatment in the same group, CD4+ T cell counts all increased at week 4, 12, and 24 in the two treatment groups, showing statistical difference (P < 0.05, P < 0.01). There was no statistical difference in the effective rate at various CD4+ T cell levels between the two groups (P > 0.05). There was no statistical difference in expression levels of Th17 and Treg cells between the combination group and the treatment group at any time point (all P >0.05). The Th17/Treg ration significantly increased in the combination group after 24 weeks of treatment, showing statistical difference when compared with the treatment group (U = 2.135, P = 0.038).

CONCLUSIONQD could improve the immune balance of Th17/Treg cells, which might be one of its mechanisms for improving HIV/AIDS patients' immunity.

Acquired Immunodeficiency Syndrome ; drug therapy ; immunology ; Adult ; Antiretroviral Therapy, Highly Active ; CD4 Lymphocyte Count ; Case-Control Studies ; Drugs, Chinese Herbal ; therapeutic use ; Female ; HIV Infections ; drug therapy ; immunology ; Humans ; Male ; Middle Aged ; Phytotherapy ; T-Lymphocytes, Regulatory ; cytology ; Th17 Cells ; cytology

OBJECTIVETo investigate the expression of microRNA-31 and its association with clinicopathologic features in oral squamous cell carcinoma (OSCC).

METHODSThe expression level of microRNA-31 in 62 cases of OSCC and matched non-tumor adjacent tissue specimens was examined using stem-loop real-time PCR. The relationship between the expression of microRNA-31 and its clinicopathologic features of OSCC was analyzed.

RESULTSThe expression of microRNA-31 was significantly higher in the tumor tissues than that in the adjacent tissues (P < 0.05).Up-regulated microRNA-31 expression was associated with the lymph node metastasis (P < 0.05) and cell differentiation (P < 0.05) in OSCC patients.No significant association was found between the expression of microRNA-31 and gender, age, lymph node metastasis, tumor size and location.Receiver operator characteristic curve (ROC) of microRNA-31 about cell differentiation resulted in a diagnostic sensitivity of 70.4% and specificity of 89.5%.

CONCLUSIONSThe up-regulated level of microRNA-31 expression may be related to the pathogenesis of OSCC.

Adolescent ; Adult ; Aged ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cell Differentiation ; Female ; Humans ; Lymphatic Metastasis ; Male ; MicroRNAs ; metabolism ; Middle Aged ; Mouth Neoplasms ; metabolism ; pathology ; Up-Regulation ; Young Adult

BackgroundThe influence of glaucoma on the quality of life in patients is of increasing concern for ophthalmologists in recent years. However,some studies demonstrated that different types of questionnaires about quality of life have various disadvantages. Therefore, to accurately and fully assess the influencing factors of quality of life in glaucoma patients is very important. ObjectiveThe present study was to survey the self-reported visionrelated quality of life(QOL) in glaucoma patients by means of utility analysis and to tentatively analyze its influencing factors. Methods Patients with glaucoma were recruited from a single tertiary ophthalmic department. Standard face-to-face interviews were conducted. Utility values of linear rating scale ( RS ) and time trade-off ( TTO ) were calculated to evaluate the self-reported vision-related QOL of the patients. The correlations of the utility values with the patients' general and ophthalmologic characteristics were also analyzed. This survey was approved by the Ethic Committee of Beijing Tongren Hospital. Oral informed consent was obtained from the subjects before the study.ResultsA cross-sectional study was designed. A total of 86 glaucoma patients were enrolled in this study with 62 male and 24 female, with a mean age of 44. 67 years old. The mean utility values measured by RS and TTO were 0. 62± 0. 19 and 0. 77 ± 0. 12, respectively, and no evidential correlation was found between these two values ( r =0. 074, P=0. 499 ). The RS value was associated with daily visual acuity,mean deviation(MD) of visual field and the history of trabeculectomy. Neither daily visual acuity nor MD showed a significant correlation with the TTO value. Age, work status and educational background contributed to higher utility value for the TTO method. After adjusting for age, work status and educational level,patients with visual acuity in the worse-seeing eye better than 0. 3 showed a higher TTO value than those with less than 0. 3. Conclusions Utility analysis possesses the advantages of convenience and sensitivity. RS utility value is easily affected by the Objective visual status and surgery history in glaucomatous patients,which reflects the subjective assessment of patients'visual quality. However, TTO value is primarily associated with age,work status and education level rather than visual function in glaucoma patients,which is therefore subjective assessment of the disease-related quality of life. These Results indicate that visual function impairment is not a determining factor for the QOL of glaucoma patients.

Objectivc To investigate the protective effect of simvastatin on retinal ganglion cells (RGC) after optic nerve crush (ONC).Methods Together 50 Kunming mice were randomly divided into normal group,sham group,ONC group and simvastatin protection group.The mice in the normal group were untreated,the sham group was treated with the exposure of the optic nerve without injury,the ONC group mice underwent ONC operation on the left eyes,followed by intravitreal administration of equilibrium solvent [50 mg · mL-1 ethanol plus 1 mol · L-1 NaOH,which were activated by 1 mol · L-1 HC1 (pH 7.2)],and the simvastatin protection group was intravitreally injected different concentrations of simvastatin (0.5 g · L-1,1.0 g · L-1,1.5 g · L-1) after ONC operation.Brn3a immunofluorescence staining,HE staining and toluidine blue staining were used to detect the apoptosis of RGC and pathological changes of optic nerve.Results On day 7 after operation,in the ONC group,the apoptosis of RGC was observed obviously,with the survival rate dropping to (35.1 ± 3.9) ％,and the thickness from the retinal ganglion cell layer to outer nuclear layer was decreased from (123.13 ± 1.04) μm to (97.48 ± 2.33) μm,which was significantly thinner than that in the control group (P ＜ 0.01);moreover,the fibrous bundle of optic nerve disappeared,and the neuroglial cells were clustered into groups,as well as the axons showed swelling and serious degeneration,but after intravitreal injection of 1.0 g · L-1 simvastatin,the survival rate of retinal ganglion cells increased to (76.3 ± 3.7) ％ (P ＜ 0.05),and the aforementioned thickness was increased to (111.39 ± 4.06) μm,which was statistically significant when compared with the ONC group (P ＜ 0.01).The degeneration of optic nerve was improved,and the structure of neuroglial cell axons and the nerve fibers became normal.Conclusion Simvastatin can reduce the optic nerve degeneration and improve the survival rote of retinal ganglion cells.

Objective To detect the expressions of hypoxia inducible factor-1α (HIF-1α) and aquaporin-1 (AQP1) in the alkali-burn cornea and investigate the roles of HIF-1α and AQP1 in alkali-burn mechanisms.Methods Totally 48 healthy adult female Kunming mice were selected,and the left eyes were treated with saline as the control group,and the model of corneal alkali burn was established with concentration of 1 mol · L-1 NaOH on the right eyes,and then these rats were randomly divided into 1-day,4-day,7-day and 14-day group.The expressions of HIF-1α and AQP1 in the cornea were detected using immunofluorescence and qRT-PCR after corneal alkali burn model was successful.Results In the control group,HIF-1 α was expressed in the corneal epithelial basement membrane,but it was increased in the corneal epithelium layer 1 day after alkali burn,and it was expressed in the corneal epithelium layer and stroma 4 days,and peaked 7 days after alkali burn;AQP1 was weakly expressed only in the endothelial layer in the control group,but it was increased in the corneal endothelial layer 1 day after alkali burn,and it was strongly expressed in the endothelial cell layer and the stroma 4 days and 7 days after alkali burn.qRT-PCR indicated that the relative expression level of HIF-1α mRNA was 269.70 ± 15.68 in 1-day group,350.50 ± 67.26 in 4-day group and 272.10 ±6.88 in 7-day group,respectively,which all were higher than that in the control group (188.70 ± 33.99),with significant difference (P ＜ 0.05).In addition,the relative expression level of AQP1 mRNA was 61.90 ± 5.45 in 1-day group and 48.34 ± 1.33 in 7-day group,which were significantly higher than that in the control group (36.43 ± 3.95),with statistical difference (P ＜ 0.05).Conclusion Alkali burn can cause the pathological changes in the cornea,and HIF-1 α and AQP1 involve in corneal injury after alkali burn.

A novel practical binary vector to get marker-free transgenic plant was constructed. The estrogen-inducible Cre/loxP DNA recombination system was adopted in this system. All non-target genes located between two identical orientation loxP sites could be excised from the transgenic genome by the Cre expression. In order to analyze this system, the target gene, GUS expression box (CaMV35s: :GUS), was inserted in the MCS outside the region franked by two loxP sites. Then it was introduced into the tobaccos. Results showed that the high-efficiency DNA recombination had take place and the target gene was working order after DNA excitation.
Base Sequence ; Genetic Vectors ; genetics ; Integrases ; metabolism ; Molecular Sequence Data ; Plants, Genetically Modified ; genetics ; Recombination, Genetic ; Tobacco ; genetics

To enhance the immuogenicity of DNA vaccines expressing the GP5 protein of Porcine Reproductive and Respiratory Syndrome Virus (PRRSV), the tegument protein VP22 (encoded by VP22 gene) of Bovine Herpesvirus 1 (BHV-1), which has been demonstrated to exhibit the unusual protein transduction property, was fused to N-terminus of GP5 of DNA vaccine construct pCI-ORF5M, resulting in pCI-VP22-ORF5M expressing VP22-GP5 fusion protein. The expression of VP22-GP5 fusion protein was confirmed by both indirect immunofluorescence assay (IFA) and Western blot. To investigate its immunogenicity, BALB/c mice were immunized with the fusion expression plasmid pCI-VP22-ORF5M and non-fusion expression plasmid pCI-ORF5M, respectively. The GP5-specific ELISA antibodies, neutralizing antibodies and lymphocyte proliferative responses were evaluated at various time points after primary immunization. The results showed that GP5-specific ELISA antibodies, neutralizing antibodies, and lymphocyte proliferative responses induced by DNA vaccine pCI-VP22-ORF5M were higher significantly than those of DNA vaccine pCI-ORF5M, indicating that fusion expression with BHV-1 VP22 significantly enhances the immuogenicity of DNA vaccine expressing the PRRSV GP5 protein, and that this strategy may also be useful to develop more efficient DNA vaccines against other pathogens.
Animals ; Antigens, Viral ; genetics ; immunology ; Artificial Gene Fusion ; Female ; Mice ; Mice, Inbred BALB C ; Porcine Reproductive and Respiratory Syndrome ; prevention & control ; Random Allocation ; Vaccines, DNA ; genetics ; immunology ; Viral Envelope Proteins ; genetics ; immunology ; Viral Structural Proteins ; genetics ; Viral Vaccines ; genetics ; immunology

Foot-and-mouth disease virus (FMDV) is the aetiological angent of a highly contagious viral disease. The complete gene encoding the structural protein of FMDV (P1) was subcloned into expression vector pGEX-KG, resulting in the fusion expression plasmid pKG-P1. After transformed into E. coli BL21(DE3) and induced by IPTG, the results of SDS-PAGE showed that the GST-P1 fusion protein was expressed in high level. The molecular weight of the fusion protein wa 110kD and the expressed products were soluble. Western-blotting was performed to confirm that the expressed fusion protein could specifically react with antiserum against FMDV. The fusion proteins were further purified by GST purification kit and an indirect ELISA (P1-ELISA) based on the purified proteins was developed. Comparison between P1-ELISA and the standard indirect haemagglutinin assay showed the two methods had 87 per cent agreement by detecting 864 serum samples, indicating the purified P1 protein was specific as the antigen of indirect P1-ELISA.
Capsid Proteins ; biosynthesis ; genetics ; immunology ; Escherichia coli ; genetics ; metabolism ; Foot-and-Mouth Disease Virus ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology

OBJECTIVETo study the relationship between HPLC fingerprint chromatogram and inhibitory effect on respiratory burst of rat PMN of leaves of crataegus L.

METHODHPLC fingerprint peaks of different species of hawthorn leaves were isolated and used for the effective experiment on the respiratory burst of rat PMN. The mathematic models of the relationship between the area and the effect of fingerprint peaks were established. According to the mathematic models, the HPLC fingerprint were change into bioactive fingerprint (include effective fingerprint and potency fingerprint) with the helps of mathematics, chemometrics, computer program simulation and etc.

RESULTThe chromatogram-effect relationship of leaves of crataegus. on respiratory burst of rat PMN was established. According to this relationship, the activities of fourteen samples of leaves of crataegus. were forecasted. It was positive correlation between the expected value and the practical value. And the correlation coefficients was 0.968 (P < 0.01).

CONCLUSIONAn all-around evaluative system, which includes not only chemical identification but also effective evaluation for traditional Chinese medicine was established. It will provide a new idea for study on fingerprint chromatogram of traditional Chinese medicine.

Animals ; Chromatography, High Pressure Liquid ; methods ; Crataegus ; chemistry ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Female ; Male ; Plant Leaves ; chemistry ; Rats ; Rats, Sprague-Dawley ; Respiratory Burst ; drug effects

OBJECTIVETo investigate the effects of testosterone on the proliferation of penile corpus cavernosal cells in male SD rats.

METHODSSmooth muscle cells (SMCs) and fibroblasts collected from the corpus cavernosal tissues of male SD rats were cultured by the enzymatic dispersion method and detected for the expression of the androgen receptor (AR) by immunohistochemistry. The effects of testosterone on the SMCs and fibroblasts were observed by methyl thiazolyl tetrazolium (MTT) assay in different concentration groups (10(-8) mol/L, 10(-7) mol/L, 10(-6) mol/L, 10(-5) mol/L, 10(-4) mol/L and 10(-3) mol/L) in comparison with the control.

RESULTSThe AR expression was found in the penile corpus cavernosal tissues. MTT assay showed that, at the concentration of 10(-5) mol/L, testosterone induced the proliferation of SMCs (68100 +/- 2200) and fibroblasts (70200 +/- 1300), with significant differences from the control ( P < 0.05), while at 10(-4) mol/L, it inhibited their proliferation (55000 +/- 1400 and 59100 +/- 1500, respectively), (P < 0.01). No significant effects were noted in the other concentration groups.

CONCLUSIONAR exists in the penile corpus cavernosal tissues of male rats. Testosterone modulates the proliferation of corpus cavernosum tissue cells through AR, and different concentrations of testosterone may be positively or negatively correlated with the proliferation of SMCs and fibroblasts.

Animals ; Cell Proliferation ; drug effects ; Cells, Cultured ; Dose-Response Relationship, Drug ; Fibroblasts ; cytology ; drug effects ; metabolism ; Immunohistochemistry ; Male ; Myocytes, Smooth Muscle ; cytology ; drug effects ; metabolism ; Penis ; cytology ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptors, Androgen ; metabolism ; Testosterone ; pharmacology