Animals ; Brain-Derived Neurotrophic Factor ; genetics ; Cells, Cultured ; Female ; Gene Expression ; drug effects ; Neurons ; drug effects ; Phenylalanine ; toxicity ; Phenylketonurias ; complications ; Rats ; Rats, Sprague-Dawley ; rho GTP-Binding Proteins ; genetics

ObjectiveTo observe the effect of Xiaoyao powder combined with prozac and psychological therapy on post-stroke depression (PSD).Methods85 PSD patients were randomly divided into the treatment group (n=43) and control group (n=42). All patients were treated with routine therapy, including prozac and psychological therapy. Patients in the treatment group were also given Xiaoyao powder. Scores of Hamilton Depression Scale (HAMD), modified Barthel index (MBI) and Scandinavian Stroke Scale (SSS) of all patients were evaluated before and after therapy.ResultsAfter treatment, the effective rate and MBI scores were significantly higher and scores of HAMD and SSS were significantly lower for patients of the treatment group compared with those of the control group (P<0.01).ConclusionXiaoyao powder combined with prozac and psychological therapy can improve depression and neural function of PSD patients significantly.

Objective To sum up the therapeutic results of 125 cases of tetralogy of Fallot(TOF),and ex- plore the optimal time and risk factors of opration,as well as perioperative management.Methods One hundred and thirth-one consecutive cases of TOF underwent corrective surgery.There were simple stenosis of infundibular portion in right ventricular outflow tract in 37 cases,stenosis of infundibulum and pulmonary valve in 14 cases,main pul- monary trunk and left/right pulmonary arteries stenosis in 74 cases,and pulmonary atresia in 5 cases.Autologousper- icardial conduit,or with waived were used for right ventricular outflow tract and right ventriculo-pulmonary artery connection.Other anomalies were corrected.Results The surgicalmortality was 4.0 %.The cause of death were se- rious low cardiac output syndrome(3 patients),respiratory function failure(1 patient),pericadial infection(1 pa- tient).Conclusion It is necessary to profonn corrective opration on younger TOF patients.Effetive prophylaxis and control of low cardiac output syndrome and pulmonary complication is a useful strategy.

AIM:To observe the improvement in the endothelial function of patients with long-term use of Shexiang Baoxin Pill(Moschus,Ginseng extract,Calculus bovis,Cortex cinnamomi,Styrax,Venenum bufonis,and Borneolum Syntheticum)(SXBXP).METHODS:Eighty patients with CHD were randomly assigned into the SXBXP group and the control group.Two groups were controlled with the usual treatment.SXBXP 2 pills,po.tid.were taken in the SXBXP group and continued for at least 6 months.The ultrasound assessment of endothelial-dependent flow-mediated vasodilation(FMD)of the brachial artery was applied in the 3rd,6th and 18th and simultaneous determination of nirtogen oxides(NO),nitricoxide synthase(NOS),superoxide dismutase(SOD),endothelin(ET)was made.RESULTS:Follow-up checkups revealed that the level of FMD and SOD in the 3rd month,the level of NO,NOS and ET in the 6th month in SXBXP group were better than those in the control group at the significant level(P＜0.05).Patient keeping on taking SXBXP up to the 18th month,the obvious improvement in the level of FMD,NO,NOS and SOD was showed(P＜0.05).CONCLUSION:Administration of SXBXP could improve endothelial function in the CHD patients significantly.

Animals ; Apoptosis ; Herpes Simplex ; physiopathology ; virology ; Humans ; MicroRNAs ; genetics ; metabolism ; Simplexvirus ; genetics ; physiology ; Virus Latency

Objective To investigate the antimicrobial resistant and transmission mechanisms of carbapenem-resistant K.pneumonia (CR-KP) infection of newborns.Methods A retrospective study was conducted on totally 37 non-repetitive CR-KP which were isolated from patients hospitalized between April 2011 and October 2013.Resistance genes were identified by PCR and sequencing.Plasmid was analyzed by pulsed-field gel electrophoresis (PFGE).Conjugation experiments were performed to determine the transferability of beta-lactamase.Multilocus sequence typing (MLST) was used to determine the genotypes and homology of these isolates.Out-membrane proteins were examined by PCR and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE).Results Thirty-seven CR-KP isolates were tested.The resistant rates of imipenem, meropenem, ertapenem were 89.2％ (33/37), 83.8％ (31/37) ,97.3％ (36/ 37), respectively.All the 37 CR-KP exhibited 100％ (37/37) sensitivity to tigecycline, colistin, levofloxacin and amikacin, while resistance to most of the other antibiotics.By PCR, 67.6％ (25/37) isolates were blaNDM-1 positive, 35.1％ (13/37) isolates were blaIMP-4 positive and 2.7％ (1/37) isolate were blaIMP-8 positive, including two isolates carrying both blaNDM-1 and blaIMP-4.PFGE results showed that the isolates carried 2-4 plasmids and both blaNDM-1 and blaIMP-4 were transferable by plasmids.MLST assigned them to sequence type (ST) 20, ST17, ST54, ST705, ST290,which showed that there were infectious outbreaks caused by NDM-1-producing and IMP-4-producing respectively among newborns.SDS-PAGE result indicated that there was no absence of outer membrane proteins OmpK35 and OmpK36.Conclusions The main resistant mechanisms of CR-KP causing infection in newborns were those the isolates carried carbapenemase of blaNDM-1 or blaIMP-4 and the K.pneumonia with two kinds of carbapemenase were detected.

Objective:To investigate the effects of shikonin on osteoclastogenesis in vitro and amelioration of bone loss in ovariectomized-induced osteoporosis in mouse model.Methods:The optimal concentration of shikonin treating were evaluated in vitro depending on its effect on the viability of C57BL/6J mouse bone-marrow-derived macrophages by CCK-8 method.To establish the osteoclastogenesis cell model,macrophages were cultured with RANKL and M-CSF treatment,and TRAP staining was used to observe the generation of osteoclasts after treating with different concentration of shikonin solution.Expressions of osteoclast marker genes,including TRAP,c-Fos and NFATclwere detected with real-time PCR.Fifthteen mice were randomly allocated into sham operation group,ovariectomized model group and shikonin treatment group.After the modeling,mice in treatment group were received the intraperitoneal injection of shikonin,while the other two groups treated with normal saline.After thirty days treatments,all animals' tibias were dissected for micro-CT analysis.Results:①The macrophages viability was significantly inhibited when the concentration of shikonin was higher than 250 nmol/(P＜0.01).②The osteoclastogenesis was significantly suppressed by differemt dose of shikonin(P＜0.01).③ The expression of the osteoclastic marker genes (TRAP,c-Fos and NFATc 1) were suppressed by addition of shikonin comparing to control group (P＜0.01).④ Shikonin effectively prevented ovariectomy-induced bone loss (P＜0.05).Conclusion:Shikonin suppresses osteoclastogenesis in vitro and ameliorates ovariectomized-induced osteoporosis in mouse model.

Objective:To investigate the mRNA and protein expressions of steroid sulfatase (STS) in breast cancer tissues and normal breast tissues, and analyze its relationship with clinicopathologic characteristics. Methods:The mRNA and protein expressions of STS, in 40 cases of breast cancer tissues and corresponding paracancerous normal breast tissues, were examined by reverse transcription-polymerase chain reaction(RT-PCR)and immunohistochemistry. The correlation of STS expression level with clinicopathologic characteristics was analyzed. Results:STS protein was mainly expressed in the cytoplasm of breast carcinoma cells and epithelial cells in normal breast glands, but not in the stroma. It could be detected in the nucleus of carcinoma cells in 3 cases of breast cancer tissues, which was pathologically classified as invasive ductal carcinoma, invasive lobular carcinoma, and invasive micropapillary carcinoma. STS was not observed in interstitial tissues of breast glands. STS protein expression had positive correlation with its mRNA expressing level. The positivity of STS was 70.0% in breast cancer tissues, significantly higher than that of normal breast tissues (42.5%). The difference was significant (P =0.013). Stratified analysis showed that the positive rates of STS protein were significantly higher in premenopausal patients, the patients with lymph node metastasis, and those with advanced breast carcinoma than those in the matched normal breast tissues (P<0.05). Conclusion:Breast cancer tissues highly expressed STS protein to stimulate local estrogen production, thereby enhancing the progression and migration of breast cancer cells. In addition, as the tumor growth, locally biosynthesized estrogens may play more and more important roles.

ABSTRACT:OBJECTIVE To investigate the role of connexin proteins (Cx), which form gap junctions (GJ), in progression and chemotherapeutic sensitivity of cervical cancer (CaCx). METHODS We analyze the expression of Cx26, Cx30, Cx32 and Cx43 in human specimens consisting of: Normal cervix (n=78), CaCx FIGO stage Ⅰ (n=148), CaCx FIGO stage Ⅱ (n=165). InCaCx cell lines, Hela- Cx32 (induced expression by doxycycline), C- 33A (endogenously express Cx32) and siHa (transiently transfected plasmid with Cx32), we detected the role of Cx32 against tostreptonigrin/cisplatin-induced apopotosisin presence or absence of functional GJ through using GJ inhibitors or low density cultural.Furtherly, we observed the relativity of Cx32 and EGFR expression in human specimens. Also, we detected the role of EGFR signaling pathway in the process of Cx32 anti-apoptosis through suppressed EGFR expression by inhibitors or siRNA sequences in cell lines. RESULTS We firstly demonstrated the expression of Cx32 was highly upregulated and accumulated in cytoplasm in the CaCx specimens, and the degree of upregulation correlated with advanced FIGO stages. Thus,in three human cervical cell lines, Cx32 was shown to suppress apoptosis when GJ formation is inhibited. No matter in cases of CaCx or cell lines, Cx32 expression was highly correlated with expression of EGFR and the EGFR pathway is an essential component of the Cx32-induced anti-apoptotic effect. CONCLUSION Cx32, traditionally tumor suppressive protein, was shown to be tumor protective against chemotherapy through EGFR pathway in a GJ-independent way.

To construct and express Hsp70-HSV2gD fusion protein. Genes of Hsp70 and HSV-2gD were subcloned into vectors pGEX-4T-1 respectively. After confirmed by DNA sequence analysis, the recombinant plasmids pGEX-4T-HSP-gD was transformed into E. coli DH5alpha and induced to express with IPTG. The expressed protein was characterized by SDS-PAGE and Western blot after purified. BALB/c mice were immunized with fusion proteins respectively via intra-m uscular injection. The proliferation of spleen lymphocytes, the level of y-IFN in culture and anti-HSV-2gD IgG antibody in serum was detected was detected. The expressed protein was analyzed by SDS-PAGE after induced with IPTG, which showed a new band with an apparent molecular mass corresponding to the predicted size (118 kD). Western Blotting analysis demonstrates that the purified Hsp70-HSV2gD fusion protein had specific binding activity. The stimulation indexes of spleen lymphocytes, the level of gamma-IFN in culture and anti-HSV-2gD IgG antibody in serum of GST-Hsp70-gD group was obviously higher than that of other groups (P < 0.05 respectively). The successful expression of the Hsp70-HSV2gD fusion protein, which can induce immune responses, laid a solid foundation for its further research.
Animals ; Blotting, Western ; Cell Proliferation ; Gene Expression ; HSP70 Heat-Shock Proteins ; biosynthesis ; genetics ; immunology ; isolation & purification ; Immunoglobulin G ; blood ; immunology ; Interferon-gamma ; blood ; Mice ; Plasmids ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; isolation & purification ; Spleen ; cytology ; immunology ; Viral Envelope Proteins ; biosynthesis ; genetics ; immunology ; isolation & purification