Objective To explore the effects ofCangfu Congxian Decoction on the mRNA expression of IGF-1R in human mural granulosa cells;To explore the action mechanism for the treatment of polycystic ovary syndrome (PCOS) patients.Methods Totally 48 patients undergoing IVF-ET were randomly divided into treatment group and control group by random number table, 24 cases in each group. Patients in the control group accepted GnRHa/FSH/hCG stimulation, while the treatment group tookCangfu Congxian Decoction orally on the basic treatment of the control group. Serum E2, LH, and P concentration on HCG injection day, number of ovum, high quality embryo rate and clinical pregnancy rate were evaluated. The mRNA expression of IGF-1R on mural granulosa cells in the mature follicle was detected by real-time PCR.Results Compared with the control group, there was no significant difference in serum E2, LH, and P concentration on HCG injection day of treatment group (P>0.05), while the treatment group had higher high quality embryo rate than that of the control group (P<0.01). There was no significant difference in the number of ovum and pregnancy rate between the two groups (P>0.05). The mRNA expression of IGF-1R on granulosa cells in treatment group was significantly higher than the control group (P<0.01).Conclusion The mechanism ofCangfu Congxian Decoction for treating PCOS is up-regulating the mRNA expression of IGF-1R in mural granulosa cells and increasing high quality embryo rate.

Objective: To investigate the loneliless situation of the elderly, and explore the relationship between loneliness,filial expectations and intergenerational social support among the elderly. Methods: Using UCLA Loneliness Scale, Intergenerational Social Support and Fealty Expectation Questionnaire, we investigated 423 samples of old people aged 60 ～94 of Changsha City. Results: The mean score of loneliness was 39.8 (s =7.3). Ordinal regression analysis indicated that the loneliness of the elderly was significantly related to received adult children social support (OR=2.208, P=0.006),afforded adult children social support(OR=2.056,P=0.01) and filial expectation(OR=1.872,P=0.027),controlled by age, sex, level of education, marriage, level of family income, and residence status. Filial expectation directly affected loneliness and indirectly affected loneliness through received adult children social support and afforded adult children social support. Conclusion: The loneliness of the elderly are significantly related to filial expectation and intergenerational support.

Objective To study the influence of sole massage on children’s analepsia from general anesthesia.Methods One hundred children in analepsia period were divided into the test group(50 cases)and the control group(50 cases).The routine nursing and monitoring during recovery stage were done in the control group.Based on the routine monitoring and nursing care as in the control group,the test group received sole massage for 10 minutes.The two groups were compared in terms of declining percentage of SpO2, rate of nausea,blood pressure,heart-rate variability,and post-analepsia dysphoria scores.Result The test group was significantly lower than the control one in all the indexes of the declining percentage of SpO2,rate of nausea,blood pressure,heart-rate variability,and post-analepsia dysphoria scores(all P<0.05).Conclusions Sole massage during children’s analepsia from general anesthesia is effective not only in lessening their declining percentage of SpO2,rate of nausea,blood pressure,heart-rate variability, and recovery dysphoria mark,but also easing their dysphoria,anxiety and fear.It may create a good condition for children to live through analepsia period successfully and safely.

Objective Up to now, there has been no sure cure for genital herpes (GH), and vaccine seems a most promis-ing approach to the prevention and treatment of herpes simplex virus Ⅱ(HSV-2) infection.In this study, we investigated the feasibili-ty of preparing a dendritic cell ( DC) vaccine modified by the adenovirus-mediated HSV-2 gD gene. Methods We subcloned the HSV-2 gD gene into the vector Shuttle-2 and constructed the recombinant adenovirus pAdeno-HSV-2 gD following identification by en-zyme digestion and DNA sequence analysis .We isolated DCs from the mouse bone marrow , analyzed their phenotypes by flow cytome-try after transfection with the recombinant adenovirus pAdeno-HSV-2 gD, and determined the expression of HSV-2 gD by immunohisto-chemistry, RT-PCR, SDS-PAGE, and Western blot. Results Based on HSV-2 DNA, the corresponding target fragments were am-plified with the gD gene primers.Agarose gel electrophoresis showed the correct size of the PCR product (1182 bp) as predicted.The recombinant adenovirus pAdeno-HSV-2 gD was obtained by transfecting the 293 cells with pAdeno-gD DNA, which had an activity of 4 ×1010 IU/mL.The contents of CD40, CD80, and CD86 were (74.2 ±3.9), (73.9 ±4.1), and (76.1 ±5.5) % in the mature DCs and (81.3 ±3.1), (80.4 ±2.9), and (83.7 ±3.9) % in the pAdeno-HSV-2 gD DCs, significantly increased as compared with those in the immature DCs ([9.7 ±0.5], [7.5 ±1.2], and [5.2 ±1.1] %) (P<0.01).No statistically significant differ-ences were found between the expression of the surface molecules in the pAdeno -HSV-2 gD DCs and that in the cytokine stimulation-in-duced mature DCs (P>0.05).RT-PCR and immunohistochemistry confirmed the expression of HSV-2 gD in DCs.SDS-PAGE and Western blot of the expressed protein showed a new band with an apparent molecular mass corresponding to the predicted size (43000). Conclusion The results of our study have paved the ground for the successful preparation and identification of a dendritic cell vaccine modified by the adenovirus-mediated HSV-2 gD gene.

Objective To screen for genes associated with low-dose UVA irradiation-induced adaptation reaction in cultured human melanocytes, and to explore the molecular mechanism of the adaptation.Methods Cultured human melanocytes of fifth to tenth passage were divided into two groups, experimental group was subjected to an irradiation with UVA at 7.2 J/cm2 once daily for 4 times and an additional irradiation at a lethal dose of 86.4 J/cm2 6 hours after the above 4-session irradiation, and control group subjected to a single irradiation with UVA at 86.4 J/cm2. A human genome-wide oligonucleotide chip was used to screen for differentially expressed genes between the two groups of cells followed by a functional classification based on international standard. Moreover, a part of these genes were analyzed and identified by real-time fluorescent quantitative PCR. Results In the adaptation reaction induced by low-dose UVA irradiation in cultured melanocytes, there were 129 differentially expressed genes, including 81 up-regulated genes and 48 down-regulated genes. These genes were found to be mainly involved in metabolism, transport, signal transduction, apoptosis, DNA synthesis and repair, and some of them were oncogenes or anti-oncogenes. Real-time PCR confirmed some of the differentially expressed genes. Conclusions The whole genome-wide oligonucleotide chip could screen with high efficiency for differentially expressed genes in low-dose UVA irradiation-induced adaptation reaction in cultured melanocytes.

AIM: To study the change of myocardial ceramide during myocardial ischemia/reperfusion and the relationship between ceramide and apoptosis and oxidative stress. METHODS: After inducing myocardial ischemia/reperfusion (I/R) injury in mice with pituitrin (Pit), myocardial SOD activity and MDA content were measured. DNA agarose gel electrophoresis and fluorescent staining of DAPI were done to check up apoptosis. The content of myocardial ceramide (?g/kg) was measured through HPTLC and scan of thin plate. RESULTS: The myocardium of I/R model group had the phenomenon of DNA ladder. Apoptosis index and ceramide content in I/R model group were higher than those in normal control group (P

Objective To establish an animal model for immunological contact urticaria in mice.Methods A total of 60 BALB/c mice were randomly and equally divided into 5 groups:anti-dinitrophenol IgE monoclonal antibody (anti-DNP IgE) + 2,4-dinitrofluorobenzene (DNFB) group and anti-DNP IgE + trimellitic anhydride (TMA) group both injected with anti-DNP IgE via tail veins firstly,followed by topical treatment with DNFB and TMA respectively on the ears at 24 hours after the injection,DNFB group,TMA group and normal saline (NS) group all injected with NS via the tail vein firstly,followed by topical treatment with DNFB,TMA and NS on the ears 24 hours after the injection.In the following 14 days,mice were observed daily for the appearance of wheals and for scratching behavior.All the mice were sacrificed at the end of the study followed by determination of the percentage of degranulated mast cells and spleen index as well as observation of pathological changes.Results Wheals were observed in all the mice (12/12) in the anti-DNP IgE + DNFB group,some mice (8/12) in the anti-DNP IgE + TMA group,but not observed in any mice in the other 3 groups.Compared with the NS group,both the anti-DNP IgE + DNFB group and anti-DNP IgE + TMA group showed a significant increase in the percentage of degranulated mast cells (70.21％ ± 26.01％ and 54.25％ ± 39.57％ vs.14.45％ ±6.79％,F=14.41,P=0.000),spleen index (7.54 ± 1.56 and 7.87 ± 1.18 vs.5.37 ± 1.16,F=4.29,P=0.004) and scratching frequency ((31.58 ± 3.58)/h and (22.17 ± 3.81)/h vs.(2.00 ± 0.85)/h at 30 minutes,F =437.86,P ＜ 0.01).Conclusion A stable mouse model for immunological contact urticaria can be established quickly by sensitization with anti-DNP IgE and challenge with DNFB.

AIM: To study the effects of Sini Decoction's effective component(SNDE) on myocardial apoptosis and ceramide content during myocardial ischemla/reperfusion. METHODS: The mice of Kunming species were randomly divided into 3 groups: control,myocardial ischemia/reperfusion and SNDE.After inducing myocardial ischemia/reperfusion injury of mice in vivo with pituitrin(Pit),myocardial SOD activity and MDA content were measured.DNA agarose gel electrophoresis and fluorescent stain of DAPI were done to check up apoptosis.The content of myocardial ceramide(?g/kg) was measured through HPTLC and scan of thin plate. RESULTS: The myocardium of model group has the phenomenon of DNA ladder.The apoptosis index and the ceramide content in model group were higher than those of control group(P

Objective To study the anti-myocardial ischemia-reperfusion injury effect of Sini Decoction's active fraction(SNDAF).Methods Experimental animals were randomly individed into normal,model,Sini Decoction(SND),and SNDAF groups.Rat hearts were perfused by the Langendorff method.The model of myocardial ischemia-reperfusion injury was reproduced by adjusting the flow of perfused liquid.The perfused liquid of normal and model groups was KH buffer saturated by oxygen.The perfused liquid with SND or SNDAF was mixed with KH buffer.The coronary flow and contract power of myocardium at 0 min of ischemia and 1,5,10,20,30,40,and 50 min of reperfusion were tested,respectively.The incidence of reperfusion arrhythmias(IRA) in the first 1 minute of reperfusion was calculated.Mice were given drugs by ig for 3 d and then myocardial ischemia-reperfusion models were established by ip pituitrin(20 U/kg).ECG of mice was recorded at 0—80 min after ip administration. SOD activity,and the contents of MDA and LA in myocardium of mice were measured.Results Both SND and SNDAF could reinforce myocardial contract and reduce the IRA in the first minute of reperfusion.SND had better effect on IRA than that of SNDAF.SNDAF had better effect on myocardial contract than that of SND.SND and SNDAF could significantly drop ECG J point raise induced by ischemia,and increase SOD activity,and decrease contents of MDA and LA significantly in ischemia myocardium.There were no significant difference between SND and SNDAF.Conclusion SNDAF could improve oxidative injury and suppress(ischemia) of myocardium,reinforce myocardial contract,and reduce the incidence of arrhythmias during(myocardia) ischemia-reperfusion.

Objective To evaluate the specific immune response induced by a dendritic cell-based adenovirus-mediated vaccine carrying the herpes simplex virus type 2 glycoprotein D gene (pAdeno-HSV-2 gD-DC) in BALB/c mice.Methods Forty BALB/c mice were equally divided into four groups:blank control group receiving no treatment,pAdeno-DC group immunized with pAdeno-DC,pAdeno-HSV-2 gD-DC group immunized with the previously constructed vaccine pAdeno-HSV-2 gD-DC,DC group immunized with DCs only.Totally,three rounds of vaccination were conducted at a 7-day interval.Ten days after the last vaccination,serum samples were collected and spleen cells were isolated from these mice.Enzyme-linked immunosorbent assay (ELISA) was performed to measure the level of IgG antibody against HSV-2 gD in the serum samples.Some spleen cells were stimulated with HSV-2 gD protein (10 mg/L) for 72 hours; then,ELISA was carried out to determine the levels of interferon (IFN)-γand interleukin (IL)-4 in the supernatant,and 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide (MTT) assay to estimate the proliferative activity of these cells.The cytotoxicity of spleen cells was also evaluated based on the measurement of lactate dehydrogenase (LDH) release.Results The serum level of IgG antibody against HSV-2 gD (given in the absorbance value at 450 nm) was 0.313 ± 0.034 in the pAdeno-HSV-2 gD-DC group,significantly higher than that in the pAdeno-DC group,DC group and blank control group (0.034 ± 0.009,0.028 ± 0.009 and 0.026 ± 0.010 respectively,all P ＜ 0.05).Increased proliferative activity and cytotoxicity were observed in spleen cells from the pAdeno-HSV-2 gD-DC group compared with those from the pAdeno-DC group,DC group and blank control group (cell stimulation index:1.600 ± 0.215 vs.1.063 ± 0.070,1.056 ± 0.063 and 1.020 ± 0.051,all P ＜ 0.05; percentage of cytotoxicity:37.1％ vs.16.0％,14.9％ and 15.7％,all P ＜ 0.05).The levels of IFN-γ and IL-4 (both given in the absorbance value at 450 nm) were 0.568 ± 0.031 and 0.544-± 0.043 respectively in the supernatant of spleen cells from the pAdeno-HSV-2 gD-DC group,compared to 0.266 ± 0.021 and 0.278 ± 0.037 respectively in the pAdeno-DC group (bothP＜ 0.05),0.271 ± 0.023 and 0.275 ± 0.044 respectively in the DC group (bothP＜ 0.05),and 0.252 ± 0.012 and 0.245 ± 0.051 respectively in the blank control group (both P＜ 0.05).Conclusion The vaccine pAdenoHSV-2 gD-DC could induce a specific and strong immune response in BALB/c mice.