Objective To study the relationship between tyrosine phosphorylation(TP)and protein expression of insulin receptor substrate-1(IRS-1)and insulin resistance in patients with gestational diabetes mellitus(GDM).Methods IRS-1 expression and TP in skeleton muscle tissue were determined by Western blot and immunoprecipitation in women with GDM(GDM group,n=22),normal pregnant women(normal pregnancy group,n=22)and normal nonpregnant women(normal nonpregnant group,n=13).Fasting plasma glucose(FPG)and fasting insulin(FINS)were measured by oxidase assay and immunoradioassay. Results(1)The levels of FPG,FINS,and insulin resistance index were calculated according to homeostasis model assessment [ HOMA-IR;(5.6?0.8)mmol/L,(15.4?5.1)mU/L,and 1.2?0.5 ] in GDM group were significantly higher than those in normal pregnancy group [(4.4+0.5)mmol/L,(10.6 ?3.1)mU/L,and 0.8?0.3;P

ObjectiveTo investigate the effects of mefformin on the levels of serum retinol-binding protein 4 (RBP-4),high sensitive C reactive protein (hs-CRP) and tumor necrosis factorα (TNF-α) in patients with impaired glucose tolerance(IGT) and metabolic syndrome(MS). MethodsSixty patients with IGT and MS were divided into mefformin treatment group (30 cases) and life-style intervention group (30 cases) by random digits table. Body mass index (BMI),the levels of HbAic, HOMA-IR, blood fat, RBP-4,hs-CRP, TNF- α were measured both before and 16 weeks after treatment in the two groups and compared.ResultsThe levels of HbA1c, HOMA-IR, RBP-4, hs-CRP and TNF- α were significantly lower in mefformin treatment group than those in life-style intervention group [(5.09 + 0.26 )％ vs. (5.69 ± 0.49 )％, 2.95 ± 0.63vs. 3.49 ± 0.78, ( 18.69 ± 6.50) mg/L vs.(26.20 ± 6.97) mg/L, (2.37 ± 0.53) mg/L vs.(2.99 ± 0.57) mg/L,(9.49 ± 2.37) μ g/L vs. ( 14.33 ± 2.62) μ g/L] (P ＜ 0.01 ). The results of multiple linear regression showed that correlations were found between the changes of RBP-4 and BMI,HOMA-IR,hs-CRP,TNF-α(β =0.284,0.506,0.274,0.230,P ＜0.01 ),and HOMA-IR was the most important limiting factor. Conclusions Mefformin can improve insulin sensitivity of the patients with IGT and MS, and depress the levels of RBP-4,hs-CRP and TNF- α. Meanwhile mefformin has anti-inflammatory effect.

SUMMARY Objective:To investigate the influence of cryoprotectants and cooling rates in vitrificationmethod on the spindles of rabbit M Ⅱ oocytes.Methods:Rabbit oocytes were verified by using cryoloopwith ethylene glycol(EG)singly or EG combined with dimethyl sulphoxide(DMSO)as cryoprotectants,and cooled by taking oocytes directly into liquid nitrogen or by vitrification machine.After frozen rabbit o-ocytes thawed,the microtubulin and chromosome of the spindles were fixation and stained by immunofluo-rescent method.Confocal microscope was used to reveal spindle configuration.Results:In the two proto-cols of single EG used and EG combined with DMSO,the spindles were severely injured.But in protocolof EG combined with DMSO and at ultra-rapid cooling rate,the normal configuration of spindle rate ofthawed rabbit oobytes was similar to that of the control group.Conclusion:The protocol of EG combinedwith DMSO as cryoprotectants and with extremely high cooling rate by vitrification machine can producethe best effect on conservation of spindle configuration in vitrification of rabbit oocytes.

The present study aimed to investigate the effect of Glomus versiforme and Trichodema harzianum on the growth and quality of Salvia miltiorrhiza continuous cropping under field conditions. The field plot experiment was conducted, these active components in the plant were analyzed by HPLC, the root diseases incidence rate of S. miltiorrhiza determined by observation and counting, and relative parameters were measured. The data was statistically processed. The result showed that inoculation of G. versiforme and combined inoculation of G. versiforme with T. harzianum significantly decreased the root diseases incidence rate of S. miltiorrhiza, and combined inoculation of G. versiforme with T. harzianum was better than other treatments. All treatments improved accumulation of active ingredients in root. Inoculation of G. versiforme and combined inoculation of G. versiforme with T. harzianum significantly increased the content of salvianolic acid B and cryptotanshinone of root (P < 0.05), Inoculation of G. versiforme, T. harzianum and combined inoculation of G. versiforme with T. harzianum significantly enhanced the content of tanshinone I and tanshinone II(A) of the root (P < 0.05). It may conclude that inoculation of G. versiforme and combined inoculation of G. versiforme with T. harzianum can effectively reduce the root diseases incidence of continuous cropping S. miltiorrhiza, and improve the quality of S. miltiorrhiza.
Antibiosis ; physiology ; Benzofurans ; metabolism ; Diterpenes, Abietane ; metabolism ; Glomeromycota ; physiology ; Host-Pathogen Interactions ; Phenanthrenes ; metabolism ; Plant Diseases ; microbiology ; Plant Roots ; growth & development ; metabolism ; microbiology ; Salvia miltiorrhiza ; growth & development ; metabolism ; microbiology ; Trichoderma ; physiology

Objective To explore immunotherapy effective combined with active immunotherapy in different time according rats bearing-tumor after paclitaxel and carboplatin chemotherapy, and to identify the optimization time and strategy of vaccine and seek rational chemo-immunotherapy strategies in ovarian cancer treatment. Methods The dynamic immunocytes number and function in established tumor treated with paclitaxel and carboplatin chemotherapy were investigated. The changes of established tumor volume and immune function of different groups were observed according to combining different time after chemotherapy and vaccine. Results Lymphopenia was observed and the number of lymphocyte subset decreased remarkably on the 6th day, but all cells were found almost recovered on 15th day after chemotherapy. There is the process of immune-enhancing from post-chemotherapy 6 day to 10 day and reversal of immune suppression temporary. The combination post-chemotherapy 6 day with CTL caused a significantly delayed tumor growth in both tumor models and induced significant the proliferation of T lymphocyte by [H] 3 releasing. The number of CD8+T cell is the highest, but the expression of Tr cell was lowest in the group of post-chemotherapy 6 day with CTL. Furthermore, the ability of CD8+T secretion IFN-γ is the most in the post-chemotherapy 6 day with immunotherapy groups. Conclusion Combinational paclitaxel and carboplatin chemotherapy has synergistic effects with active immunotherapy boosting against tumor during window periods, where 6 days after chemotherapy with the most decreased number of lymphocytes in the animal periphery might represent the optimal checkpoint for the immune therapy against tumors. Therefore, monitoring the immune status of tumor patients might become one of the important prerequisites for the effective immune therapy when designing the comprehensive therapeutic strategies.

Objective To investigate the clinical spectrum of respiratory viruses in infants and young children with acute lower respiratory infection(ALRI) in Chongqing area from 2003-2007.And to assess the clinical diagnostic value of virus detection in nasopharyngeal secretions(NPS) and serum viral antibody detection for ALRI.Methods Cases of 2 529 specimens of NPS in hospitalized children with ALRI from Apr.2003 to Oct.2007 were taken for detecting 7 common respiratory virus antigens by immunofluorescence assay including respiratory syncytial virus (RSV),adenovirus(ADV),influenza A(IA),influenza B (IB),parainfluenza virus1-3 (PIV1,PIV2,PIV3).Fifty-five thousand eight hundred and eighty-seven samples were tested for ADV-IgM by ELISA.Among those,45 159 cases were further tested for RSV-IgM by ELISA.Results Respiratory virus pathogens were detected in 778 samples out of 2 529(30.76%) including RSV positive in 668 samples (85.86%),PIV3 positive in 75 samples (9.64%),IA positive in 22 samples (2.57%),ADV positive in 15 samples ( 1.93%),only 1 sample ( 0.13%) positive for both PIV1 and RSV. And the positive rate of RSV-IgM was 0.9%-15.2%,and the positive rate for ADV-IgM was about 0.6%-10.6%.RSV infection occured mainly in winter and spring.Conclusions Respiratory virus is the most common pathogen in children with ALRI during the survey period in Chongqing area,especially for RSV infection.The pattern of RSV circulation varied every year with seasonality.It is suggest that this year is peak one for RSV infection from the monthly positive results,especially in Feburary(50%) in 2007.But the infection rate of PIV3,IA,ADV and PIV1 are lower,particularly IB and PIV2 infection have not been seen for the last 5 years.It is fast and accurate to detect RSV antigen and suit to clinical diagnosis by using immunofluorescence assay than other antibody detection.

Objective To evaluate the levels of oxidative stress in patients undergoing long-term and repetitive exposure to hyperbaric oxygen (HBO) treatment. Methods 16 healthy volunteers and 58 patients with sub-acute sudden hearing loss (SHL) exposed to HBO were included in the study. Oxidative stress indices (malondialdehyde, MDA, advanced oxidation protein products, AOPP; superoxide dismutase, SOD) were measured in peripheral blood samples collected at the 5th,10th, 20th and 30th HBO treatments sessions (PO2 0.18 MPa, 1 session per day and 5 sessions per week) and under normal ambient pressure respectively. Results After 5th,10th, 20th and 30th sessions of HBOT, no relevant differences in these three indices were detected compared to pre-HBO exposure, between healthy volunteers (P > 0.05). Conclusions The long-term repetitive HBO treatment for 0.18 MPa of PO2 and 30 sessions could not affect in particular the response of the oxidative stress in healthy persons and patients with sub-acute SHL. The influence on three indices of patients with abnormal situation of oxidative stress undergoing lower pressure of HBO (0.18 MPa) is under investigation.

AIM: To construct a recombinant adenovirus vector carrying AFP promoter to specifically express a targeting gene in hepatocellular carcinoma HepG2 cells. METHODS: Based on the Adeno-X~TM Expression system, the CMV promoter was replaced by a 300 bp a-fetoprotein promoter. The EGFP(enhanced green fluorescent protein) gene as a report gene was inserted to the multiple-cloning site(MCS). The normal liver LO2 cells, hepatocellular carcinoma HepG2 cells and HeLa cells were infected by the recombinant adenovirus, respectively. Northern blotting and fluorescence microscope were used to detect the transcription level of EGFP gene and its protein expression, respectively. RESULTS: Northern blotting showed that the target gene was markedly transcribed in HepG2 cells, but slightly in LO2 and HeLa cells. Under the fluorescence microscope, strong EGFP expression was seen in HepG2 cells but very weakly in HeLa and LO2 cells. CONCLUSION: Under the control of the 300 bp human AFP promoter, the target gene carried by the recombinant adenovirus was expressed in the AFP-producing HepG2 cells at a very high level, but not or very weakly in AFP negative cells. This adenovirus system can be used as a new, potent and specific approach for the gene-targeting therapy for the AFP producing primary hepatoma. [

We designed and synthesized eighteen lycorine derivatives with five different structural types, and evaluated their antiviral activities on a HCoV-OC43-infected H460 cell model. Structure-activity relationships suggested that the introduction of appropriate substituents on the 6N atom of lycorine was beneficial to activity. Compound 6a gave a good activity with the half effective concentration (EC₅₀) and selectivity index (SI) values of 2.36 μmol·L^-1 and 16.52, respectively. Surface plasmon resonance (SPR) result indicated that 6a might target the non-structural protein 12 (NSP12) subunit in RNA-dependent RNA polymerase (RdRp) of SARS-CoV-2 with the dissociation constant (K_D) value of 1.36 μmol·L^-1. Molecular docking indicated that 6a might act on nidovirus RdRp-associated nucleotidyltransferase (NiRAN) catalytic center of NSP12, distinct from the mechanism of nucleoside-like drugs such as remdesivir. This study provides scientific data for the development of lycorine derivatives into a new class of anti-SARS-CoV-2 small molecule inhibitors.

Sini decoction (SND), a classical traditional Chinese medicine emergency formula recorded in Shanghan Lun (Treatise on Febrile Diseases), which is composed of Aconiti Lateralis Preparata Radix, Glycyrrhizae Preparata Radix and Zingiberis Rhizoma. Modern clinical and pharmacological researches have shown that SND can protect the myocardium effectively during myocardial ischemia reperfusion injury (MI-RI). A myocardial ischemia reperfusion injury model of H9c2 cardiomyocytes in vitro has been established. Four groups, control group, MI-RI Model group, SND group and SND without Glycyrrhizae Radix group, were arranged. The livability, the level of LDH and CK activity in H9c2 cardiomyocytes in different groups were tested. By combining with principal components analysis (PCA), partial least squares discriminant analysis (PLS-DA), orthogonal partial least squares projection of latent structures-discriminant analysis (OPLS-DA), 17 biomarkers in extracellular fluid were identified and 15 of them were related to the pathway of biological processes. The results showed that the attenuate and synergistic mechanism between Aconiti Lateralis Praeparata Radix and Glycyrrhizae Radix for toxicity reduction was related with the glycolysis, lipid metabolism, citrate cycle and nitrogen metabolism of amino acids metabolism. The study proved the effect on H9c2 cardiomyocyte treated by MI-RI injury both SND group and SND without Glycyrrhizae Radix group, and compared with the SND without Glycyrrhizae Radix, the protective effect of myocardial ischemia reperfusion injury model of H9c2 cardiomyocyte from SND was stronger.
Aconitum ; chemistry ; Animals ; Drug Synergism ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Glycyrrhiza ; chemistry ; Humans ; Male ; Metabolomics ; Myocardial Reperfusion Injury ; drug therapy ; genetics ; metabolism ; Myocytes, Cardiac ; drug effects ; Rats, Sprague-Dawley