Objective To investigate the effects of 1,25-dihydroxyvitamin D_3 on the bone metabolism of primary osteoporosis.Methods Female New Zealand white rabbits aged 8 months were ovariectomized bilaterally as models of postmenopausal osteoporosis and they were randomly divided into 4 groups: pseudo-ovariectomized group(Sham group),ovariectomized group(OVX group),calcii gluconas group(OC group),calcii gluconas and 1,25-dihydroxyvitamin D_3 group(OCR group).Both female New Zealand white rabbits aged 3 years and male New Zealand white rabbits aged 4 years were selected as models of senile osteoporosis.They were randomly divided into 3 groups:control group,calcii gluconas group(Calcium group),calcii gluconas and 1,25-dihydroxyvitamin D_3 group(CR group).Rabbits in OC group and Calcium group were given calcii gluconas and in OCR group and CR group were given calcii gluconas and 1,25-dihydroxyvitamin D_3.The bone metabolic biochemical indexes were determined among all the experimental animals after they were given drug for 8 weeks.Results After the experimental animals were given drug for 8 weeks,the serum calcium(Ca),serum phosphorus(P) and alkaline phosphatase(ALP) of OCR group were significantly higher than those of OVX group and OC group(all P

Objective To analyze the correlation of Brown attention deficit disorder scale(BADDS)parent form and Conners parent ratting scale(CPRS)in Chinese children ages 8-12.Methods Both BADDS parent form and CPRS on 146 children ages 8-12 in an elementary school in Xicheng district in Beijing were admmistered,and the results were compared with statistic methods.Results Total scores on the BADDS parent form were highly correlated with CPRS index scores(r=0.739,0.771 Pa

Objective To observe the immunologic function of critically ill newborn and the relative function of nuclear factor kappa B(NF-?B).Methods The critically ill group contained 50 cases,and 25 cases from healthy newborns were used as control group.Blood samples were collected in each case,levels of cytokine interleukin(IL)-4,interferon(IFN)-?,tumor necrosis factor(TNF)-? and NF-?B were detected.Result Compared with control group,NF-?B of the critically ill newborn activated and the cytokine were disorder,and IL-4 and TNF-? increased,but IFN-? decreased.Conclusions Critically ill newborn exist immune functional disorder.Furthermore,NF-?B activation may be involved in the process in infants with critically illness.

OBJECTIVETo determine the regulatory effects of the circadian clock gene Per1 on cell cycle-related genes and its influence on the proliferation, apoptosis, cycle, and tumorigenicity in vivo of human oral squamous cell carcinoma SCC15 cells.

METHODSThree groups of the short hairpin RNA (shRNA) of lentivirus recombinant plasmids were designed against the RNA of Per1 and then transfected to the SCC15 cells. The optimum interference group was screened through Western blot and quantitative real-time PCR (qRT-PCR) and assigned as the experimental group. The transfected lentivirus plasmid without an interference effect on any gene was set as the control group (Control-shRNA). Untreated SCC15 cells were set as the blank group. The mRNA expressions of cell cycle-related genes, namely, Per1, p53, Cyclin D1, Cyclin E, Cyclin A2, Cyclin B1, CDK1, CDK2, CDK4, CDK6, p16, p21, Wee1, cdc25, E2F, and Rbl1 in each group were detected through qRT-PCR. The cell proliferation, apoptosis, and cell cycle distribution in each group were evaluated through flow cytometry. The cells of the experimental group and the blank group were subcutaneously inoculated in nude mice to observe tumorigenesis.

RESULTSThree groups of Per1-shRNA lentivirus plasmids were constructed successfully. Among the groups, the Per1-shRNA- I group exhibited the highest interference effect, as indicated by qRT-PCR and Western blot analysis. As such, this group was set as the experimental group. The mRNA expression levels of CyclinD1, CyclinE, CyclinB1, CDK1, and Wee1 gene in the Per1-shRNA-I group were significantly higher than those in the Control-shRNA group and the SCC15 group (P < 0.05). By contrast, the mRNA expression levels of p53, Cyclin A2, p16, p21, and cdc25 in the Per1-shRNA-I group were significantly lower than those in the Control-shRNA group and the SCC15 group (P < 0.05). The mRNA expression levels of each gene between the Control-sLRNA group and the SCC15 group did not significantly differ (P > 0.05). The mRNA expression levels of CDK2, CDK4, CDK6, E2F, and Rb1 did not significantly differed in the three groups (P > 0.05). The proliferation index of the Perl-shRNA-I group was significantly higher than those of the Control-shRNA group and the SCC15 group (P < 0.05). The apoptosis index of the Per1-shRNA-I group was significantly lower than those of the Control-shRNA group and the SCC15 group (P < 0.05). The number of S-phase cells in the Per1-shRNA-I group was significantly lower than those of S-phase cells in the Control-shRNA group and the SCC15 group (P < 0.05). The number of G2/M-phase cells in the Per1-shRNA-I group was significantly higher than those of G2/M-phase cells in the Control-shRNA group and the SCC15 group (P < 0.05). Conversely, the proliferation index, apoptotic index, and cell cycle distribution of the cells in the Control-shRNA group did not significantly differ from those of the SCC15 group (P > 0.05). The tumorigenic ability in vivo was significantly enhanced in the Per1-shRNA-I group (P < 0.05).

CONCLUSIONPer1 is an important tumor suppressor gene. Per1 can regulate a large number of downstream cell cycle-related genes. The alteration of its expression can affect cell cycle progression, proliferation, apoptosis imbalance, and tumorigenic ability in vivo. Further studies on Per1 may elucidate cancer development and provide novel effective molecular targets for cancer treatment.

Animals ; Apoptosis ; Carcinoma, Squamous Cell ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Circadian Clocks ; genetics ; Cyclin D1 ; Humans ; Mice ; Mice, Nude ; Mouth Neoplasms ; Period Circadian Proteins ; genetics ; Plasmids ; RNA, Small Interfering ; Real-Time Polymerase Chain Reaction ; Transfection

Aged ; Carcinoma, Squamous Cell ; metabolism ; pathology ; surgery ; Humans ; Laryngeal Neoplasms ; metabolism ; pathology ; surgery ; Male ; Membrane Proteins ; metabolism ; Neoplasm Recurrence, Local ; Reoperation ; Tumor Suppressor Protein p53 ; metabolism ; Vimentin ; metabolism

Adult ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Hepatitis B, Chronic ; complications ; drug therapy ; Humans ; Liver Cirrhosis ; drug therapy ; etiology ; Male ; Middle Aged ; Phytotherapy ; Tablets

Objective To investigate the methods of isolation, cultivation of adipose derived stem cells (ADSCs) from the rats'adipose tissue and the feasibility of inducting differentiation to chondrocyte.Methods The adipose tissue was obtained from the wistar's male rats, and it was isolated, cultivated, and subcultured. The expression of CD29 in the subcultured ADSCs was detected by immunofluorescence. The 3rd generation ADSCs was cultured in the medium contained TGF-β, and the presence was identified by type Ⅱ collagen immunohistochemistry. Results The character of ADSCs'morphology could be observed by microscope, and it can be identified by immunofluorescence. Through the course of inducting the ADSCs to chondrocyte, the feasibility of inducting ADSCs to chondrocyte showed that type Ⅱ collagen was expressed.Conclusion We demonstrated the presence of stem cells in the adipose tissue, and the stability of biological feature in vitro. The chondrocyte could be obtained from the adipose tissue through the committed differentiation, induction, cultivation by exogenous transforming growth factor (TGF-β). The ability of the adipose stem cell differentiating to chondrocyte was also proved.

Objective To evaluate the value of nuchal translucency (NT) thickness in the fetal chromosome abnormality screening. Methods 11 086 pregnant women received NT measurement in 11 ~ 13+6 weeks at Hainan general hospital from January 2010 to December 2014 were selected in the study. The NT thickness was measured according to guidelines from Fetal Medicine Foundation. 122 fetuses (NT≥2.5 mm) were recruited to accept karyotype analysis. Results 11 086 pregnant women received NT measurement in 11 ~13+6 weeks, in which 122 cases′ NT are more than or equal to 2.5 mm, with a positive rate of 1.10%. 122 cases of fetal NT thickening are between 2.5 to 12.0 mm, with the average degree at (4.5 ± 2.1)mm. 122 invasive prenatal diagnostic specimens chromosome karyotype analysis results showed chromosomal abnormalities in 21 cases (abnormal rate of 17.2%), the abnormal chromosome number in 17 cases and abnormal structure in 4 cases. The top 3 are trisomy 21 (12 cases, 57.1%), chromosome pericentric inversion (3 cases, 14.3%), and trisomy 18 (2 cases, 9.5%). Fetal chromosomal abnormalities resulting from different childbirth age, the sex of the fetus, NT thickness showed significant statistical difference (P < 0.05). The concrete manifestation is that fetal chromosomal anomaly detection rate in childbirth by women more than 35 years old age are higher than other age. Female fetal chromosomal anomaly detection rate is higher than the male , and NT thickness of 5mm of fetal chromosomal abnormality rate is significantly higher than the thickness of NT group at 2.5mm~ and 3.5mm~. Fetal NT thickening of NT measurements was in significant positive correlation with fetal chromosome abnormal rate (χ2=15.533, P < 0.001). Logistic regression analysis found that with a higher NT thickness , risk of fetal chromosomal abnormalities would be significantly higher , and thickening of NT could be an independent predictor of fetal chromosome abnormalities. Conclusion In early pregnancy, ultrasound examination of fetal ultrasound screening of NT thickness can be used as an important index of fetal chromosomal abnormality , and interventional diagnosis of prenatal NT thickness increase could pose increased risk of fetal chromosomal abnormalities.

Objective To investigate the effect of construct the Notch1 (NICD) eukaryotic expression vector on the proliferation and differentiation of rat bone marrow mesenchymal stem cells (BMSCs) in vitro. Methods Rat BMSCs were experimented as the object. NICD eukaryotic expression vector was constructed. pEGFP-N1-NICD expressing plasmids were used to transfect BMSCs. The study included control group (CON group), empty vector group (VEC group) and the trans-fection group (TRA group). After 48-hour transfection, BMSCs were observed for general morphology. The protein expres-sions of NSE, GFAP and Notch1 were detected by real-time PCR and Western blotting assay respectively. The apoptosis, cy-cle distribution and cell proliferation were evaluated by flow cytometry and MTT assay. Results The DNA sequencing con-firmed that the pEGFP-N1-NICD recombinant plasmid was successfully constructed, and both VEC group and TRA group expressed green fluorescence after 48-hour transfection. The relative expression levels of Notch1 and GFAP mRNA and pro-tein were significantly higher in TRA group than those in VEC group and CON group (P<0.05), and there was no significant difference between VEC group and CON group. After 48-hour transfection, the ratio of living cells was significantly lower in TRA group than that of CON group and VEC group, and the early apoptotic rate and late apoptotic rate were significantly higher in TRA group than those of CON group and VEC group (P<0.05). The late apoptotic rate was significantly higher in VEC group than that of CON group. The proportion of G1/G0 cells was significantly higher in TRA group than that of CON group and VEC group, but S and G2/M cells were significantly lower (P<0.05). The value of growth curve was gradually de-creased in TRA group than that of CON group and VEC group (P<0.05). Conclusion The high expression of NICD gene might induce apoptosis of BMSCs, inhibit the proliferation in part, and induce into glial-like cell differentiation.

Objective To investigate the effects of dedicator of cytokinesis 1 (Dock180) knockout on proliferation and apoptosis in rat derived H9C2 cardiomyocytes and their mechanisms.Methods A single guide RNA (sgRNA) targeting rat Dock180 gene was designed and constructed using CRISPR/Cas9 system.A plasmid contained above sgRNA was packaged into lentivirus and selected to knockout Dock180 in the cardiomyocytes.A single clone of cardiomyocyte with Dock180 knockout was established.Cardiomyocytes were divided into negative lentivirus group (Cas9, A group), Dock180 knockout group (B group), Cas9 lentivirus hypoxia group (C group), Dock180 knockout hypoxia group (D group).The expression of Dock180 mRNA was examined by RT-PCR, and relevant proteins were detected by Western blot.The cell proliferation rate of the cardiomyocytes was determined by MTT, and the apoptotic rate was measured by flow cytometry.Results Dock180 mRNA and protein were absent in B andD groups.Compared with A and C groups, p-ERK1/2 and Bcl-2 protein expression and cell proliferation rate were lower in B and D groups respectively (P<0.01), while Bax protein expression and cell apoptosis rate were higher in B and D groups respectively (P<0.05, P<0.01);Compared with A group, Dock180 mRNA and protein, p-ERK1/2 and Bcl-2 proteins and cell proliferation rate were reduced, while Bax protein and cell apoptosis rate were increased in C group(P<0.05,P<0.01).Compared with B group, p-ERK1/2 and Bcl-2 proteins and cell proliferation rate were decreased, while Bax protein and cell apoptosis rate were increased in D group(P<0.05,P<0.01).Conclusions Dock180 knockout with CRISPR/Cas9 can inhibit proliferation and promote apoptosis via p-ERK1/2, Bcl-2 and Bax in H9C2 cardiomyocytes.