Administration, Rectal ; Adolescent ; Adult ; Drugs, Chinese Herbal ; administration & dosage ; Endometriosis ; drug therapy ; Female ; Humans ; Middle Aged ; Phytotherapy

Objective To study the expression and clinical significance of Survivin and PTEN in cervical carcinoma. Methods The expression of Survivin and PTEN in 60 cases of cervical carcinoma, 15 cases of CIN Ⅰ , 15 cases of CIN Ⅱ, 15 cases of CIN Ⅲ and 15 cases of normal cervical tissues were detected by immunohistochemical staining. Results The expression positive rates of Survivin were 86.67 %, 80.00 %,33.33 %, 20.00 % and 0, respectively, and those of PTEN were 21.67 %, 40.00 %, 80.00 %, 86.67 % and 100.00 % in cervical carcinoma, CIN Ⅲ, CIN Ⅱ, CIN Ⅰ and normal cervical tissues, respectively. The expression positive rates of Survivin were increased along with normal cervical epithelium, CIN Ⅰ, CIN Ⅱ,CIN Ⅲ and invasive carcinoma of cervix;while those of PTEN were inverse order.Compared with the expression positive rates of Survivin and PTEN protein in the CIN Ⅱ group, those in the CIN Ⅲ group had significant differences (P <0.01 and P <0.05, respectively). Expressions of Survivin and PTEN were correlated to the size of focus(P <0.05), the depth of tumor and pelvic lymphnode metastasis (P <0.05), but not to clinical staging, pathological types, pathological grading and age(P >0.05). There was a negative correlation between the expression of PTEN and Survivin in cervical carcinoma. Conclusion The expressions of Survivin and PTEN are correlated with invasion and metastasis of cervical carcinoma, and be related to clinical pathotology.Survivin and PTEN may play a important role in the formation, proliferation, differentuation and metastasis of cervical carcinoma. They may be used as markers of early diagnose, efficacy and prognosis evaluation.

10.3969/j.issn.1000-8179.2013.12.005

Objective To summarize the clinical and laboratory characteristics of patients with severe fever with thrombocytopenia syndrome (SFTS ) and to identify the related risk factors for mortality .Methods Clinical features and laboratory parameters were collected from 40 SFTS patients (7 deaths and 33 survivors) .Dynamic changes of laboratory data were compared between the two groups , including white blood cell count (WBC ) , platelet count (PLT ) , alanine aminotransferase (ALT ) , aspartate aminotransferase (AST) ,creatine kinase (CK) ,lactate dehydrogenase (LDH) ,prothrombin time (PT) ,activated partial thromboplastin time (APTT) and thrombin time (TT) .Continuous variables with normal distribution were compared with t test ,and those with non‐normal distribution were compared with nonparametric test ;categorical variables were compared with χ2 test .Univariate Logistic regression was used to evaluate the risk factors associated with death .Results For the deceased patients and the survivors ,the APTT were 56 .40 s and 44 .45 s ,respectively (Z=5 .419 ,P=0 .04) at day 1—7 .Those were 66 .25 s and 36 .85 s ,respectively (Z=10 .112 ,P=0 .009) at day 8—10 ,and (125 .06 ± 11 .88) s and (33 .44 ± 6 .50) s ,respectively (t=45 .760 ,P<0 .01) at day 11—13 .At day 11—13 ,the ALT levels in deceased patients and survivors were (783 .00 ± 210 .12) U/L and (137 .33 ± 89 .59) U/L ,respectively (t=7 .989 ,P=0 .016) ,AST levels were 890 U/L and 99 U/L ,respectively (Z=60 .248 ,P <0 .01) , CK levels were 2 315 U/L and 314 U/L ,respectively (Z= 122 .065 , P< 0 .01) ,LDH levels were 1 075 U/L and 509 U/L ,respevtively (Z=44 .642 ,P<0 .01) ,PT were 16 s and 11 s ,respectively (Z=7 .917 ,P=0 .031) ,and TT were 120 s and 20 s ,respectively (Z=1 361 .674 ,P<0 .01) .Day 11—13 after the onset of illness was the critical stage for SFTS .Consciousness alteration (OR=6 .60 ,95% CI:2 .94—14 .80) ,bleeding (OR=9 .29 ,95% CI:1 .48—58 .47) ,PT> 15 s (OR= 24 .00 ,95% CI:1 .99—289 .60) ,APTT>70 s (OR= 42 .67 ,95% CI:3 .54—514 .85) and TT > 120 s (OR= 0 .14 ,95% CI:0 .02—0 .88) were risk factors for the death of SFTS patients (all P< 0 .05) .Conclusion Prolonged APT T ,T T and PT at early stage and progressively increasing during the disease course suggest poor prognosis of SFTS .

Objective To determine chromosomal localization of the primary gout susceptibility gene in a pedigree.Methods The clinical data and the peripheral blood samples were collected in the pedigree members and the genomic DNA was extracted from peripheral blood.A genome-wide screening was performed using 400 micro-satellite DNA markers in this family,and linkage analysis was used to determine the chromosomal location of the primary gout susceptibility gene.Results Linkage analysis showed that the maximum LOD score reached 1.50 at marker D4S1572 (at recombination fraction?=0.00).Conclusion Since D4S1572 is localized at 4q25,the primary gout susceptibility gene of this pedigree is localized at 4q25.

Objective To observe the expression of cardiotrophin-1(CT-1) in ischemia-reinfusion cardiac muscle of rats and the effect of neuregulin-1(NRG-1).Methods The model of ischemia-reinfusion cardiac muscle of rats were prepared,35 rats were randomly divided into 4 groups:model group(n=8),NRG-1 pretreatment group(n=9),pseudo-surgery group(n=8) and normal control group(n=10).The CT-1 mRNA in the observed cardiac muscle of all groups was measured by RT-PCR and the relative amount of CT-1 mRNA were calculated,and for statistical treatment.Results The CT-1 mRNA of model group was(63.96?9.34),and it was higher than that of pseudo-surgery group(36.16?5.43)and normal control group(36.84?4.64).The significant differences were found in 3 groups(F=47.37 P

Objective To investigate the protective effect of carnosine on Glutamate induced apoptosis in SH-SY5Y cells and its mechanism. Methods The injury model was established by treating SH-SY5Y cells with glutamate in vitro.Inverted microscope was used to observe cell morphology. The cell viability was detected by MTT assay, and changes in nucleus were detected by Hoechst33258 staining under fluorescence microscopy.The cell apoptosis rates were measured by flow cytometry.The reactive oxygen species ( ROS ) level in cells was detected by fluorescent probe CDFH-DA. Results Compared with normal control group, the cell viability in glutamate group decreased (P<0.01), the morphology changes of cell apoptosis such as karyopyknosis and split were observed by Hoechst33258 staining, while the apoptosis rate and the level of ROS were increased (P<0.01). Compared with glutamate group, the cell viability of carnosine 0.6,3,15 mmol/L groups obviously increased(P<0.01), while the morphology changes of cell apoptosis significantly improved, the apoptosis rate and the level of ROS were obviously reduced ( P<0.01 ) .Compared with normal control group, the cell viability, the morphology changes and the apoptosis rate did not significantly change in carnosine group.The level of ROS was reduced, but there was no statistically significant difference between two groups.Conclusion Carnosine has apparent protective effect on apoptosis of SH-SY5Y cells injury induced by glutamate.The mechanism may be related to carnosine prevent the occurrence of oxidative stress.

OBJECTIVETo analyse a special kind of Schisandra chinensis with the white fruit using ITS2 barcode at molecular levels.

METHODITS2 regions were sequenced bidirectionally. Sequence assembly and consensus sequence generation were performed using the CodonCode Aligner, MEGA 5.0 software was used to align the sequences. The ITS2 secondary structure was predicted using ITS2 web server, BLAST 1 method was used to identify the S. chinensis with the white fruit.

RESULTThe length of the ITS2 sequence was 231 bp. And the sample was identified as S. chinensis using the method of BLAST 1. Their mean interspecific genetic distance (K2P distance) among the populations of the S. chinensis with the white fruit and S. chinensis was far lower than the mean interspecific genetic distance between the S. chinensis and S. sphenanthera.

CONCLUSIONBy using ITS2 the S. chinensis with the white fruit was identified as S. chinensis, and the ITS2 barcode could be used to identify S. chinensis and S. sphenanthera.

DNA, Plant ; chemistry ; genetics ; DNA, Ribosomal Spacer ; chemistry ; genetics ; Fruit ; chemistry ; classification ; genetics ; Molecular Sequence Data ; Nucleic Acid Conformation ; Schisandra ; chemistry ; classification ; genetics ; Sequence Analysis, DNA ; Software

Objective To investigate the protective effects of lentivirus mediated Bcl-2-associated athanogene 1L (BAG-1L) over-expression on human neuroblastoma cells (SH-SYSY) induced by hypoxia/re-oxygenation,and to study its effect on the phosphoinositide 3 kinase serine/threonine protein kinase (PI3K/AKT) pathway.Methods SH-SYSY cells were cultured in vitro,and the cells at logarithmic phase were collected,and they were divided into recombined lentiviral infection group [infected by lentivirus containing BAG-1L and green fluorescent protein (GFP) gene],vector control group (infected by lentivirus containing GFP without BAG-1L gene) and cell control group (non-infection).Western Blot was used to detect the expression of BAG-1L in target cells after infection for 48 hours.SH-SY5Y cells were subjected to hypoxia for 8 hours and re-oxygenation for 24 hours,then the cell counting kit-8(CCK-8) was used to detect the cell activity,and the apoptosis was detected by flow cytometry after allophycocyanin labeled annexin V/7-amino actinomycin D (Annexin V-APC/7-AAD) staining.Western Blot was used to detect the protein expressions of BAG-1L,heat shock protein 70 (HSP70),AKT and phosphorylated AKT (p-AKT).Results After infection for 48 hours,exogenous BAG-1L protein bands were observed in recombined lentiviral infection group,but not observed in cell control group and vector control group.After hypoxia/re-oxygenation treatment,the cell viability in recombined lentiviral infection group was significantly higher than that in cell control group and vector control group (A value:0.689 ± 0.036 vs.0.425 ± 0.013,0.400 ± 0.012),apoptosis was significantly decreased [apoptosis rate:(26.97 ± 1.82)％ vs.(36.60± 1.45)％,(35.77 ± 3.74)％],the protein levels of BAG-1L,HSP70 and p-AKT were significantly increased [BAG-1L protein (gray value):2.405 ± 0.167 vs.0.529 ± 0.141,0.601 ± 0.099;HSP70protein (gray value):0.997±0.123 vs.0.634±0.091,0.584±0.106;p-AKT protein (gray value):1.234±0.118 vs.0.661 ± 0.210,0.712 ± 0.199,all P ＜ 0.01],but the protein level of AKT was slightly increased (gray value:1.103 ± 0.269vs.0.646 ± 0.188,0.791 ± 0.326) without statistically significant differences (both P ＞ 0.05).There was no significant difference in all parameters between cell control group and vector control group (all P ＞ 0.05).Conclusion Lentivirus mediated BAG-1L gene over-expression can protect nerve cells against hypoxic injury and apoptosis,and the protective effect may be related to the activation increase of pathway on PI3K/AKT.

Objective To investigate the effects of lentivirus-mediated heat shock protein 70 (HSP70) gene on voltage-gated calcium channels on the cell membrane of pheochromocytoma cell 12 (PC 12 cells) induced by ischemic/hypoxia and its mechanisms.Methods PC12 cells at logarithmic phase were collected,which were challenged with hypoxia for 6 hours and reoxygenation for 12 hours to stimulate the nerve cells suffered ischemia/reperfusion pathological process in vitro.PC12 cells were divided into non-infection group,infected by lentivirus containing green fluorescent protein (GFP) without HSP70 gene lentivirus control group (GFP lentivirus control group),and infected by lentivirus containing HSP70 and GFP gene recombined lentiviral infection group (HSP70 recombined lentiviral infection group).Real-time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western Blot were used to determine mRNA and protein expressions of L-type voltage-gated Ca2+ channel subunits cav1.2 and cav1.3,receptor gated channel subunits NR1 and NR2,and Na+/Ca2+ exchange (NCX) in PC12 cells.Results After being challenged with hypoxia/reoxygenation,the mRNA and protein expressions of cav1.2,NR1 and NR2 in the PC12 cells were significantly lower in HSP70 recombined lentiviral infection group than those of GFP lentivirus control group and non-infection group [cav1.2 mRNA (2-△△Ct):3.13 ± 0.46 vs.5.12 ± 0.52,5.13 ± 0.66;NR1 mRNA (2-△△△Ct):1.61 ± 0.44 vs.3.23 ±0.82,3.31 ±0.78;NR2 mRNA (2-△△Ct):2.09±0.41 vs.3.91 ±0.64,3.88±0.62;cav1.2 protein (gray value):2.82±0.39 vs.3.98±0.23,3.96±0.24;NR1 protein (gray value):1.84±0.35 vs.2.79±0.21,2.86±0.23;NR2 protein (gray value):0.87±0.24 vs.1.57±0.31,1.33±0.44;all P ＜ 0.01].But there were no statistical differences in the mRNA and protein expressions of cav1.2,NR1 and NR2 between GFP lentivirus control group and non-infection group (all P ＞ 0.01).There were no statistical differences in the mRNA and protein expressions of cav1.3 and NCX among non-infection group,GFP lentivirus control group and HSP70 recombined lentiviral infection group [cav1.3 mRNA (2-△△Ct):4.82 ± 0.32,4.72 ± 0.36,4.82 ± 0.29;NCX mRNA (2-△△Ct):3.49 ± 0.78,3.47 ± 0.71,3.56 ± 0.65;cav 1.3 protein (gray value):2.63±0.40,2.64±0.39,2.68±0.39;NCX protein (gray value):3.27±0.48,3.34±0.48,3.31 ±0.42;all P ＞ 0.01].Conclusion Exogenous HSP70 affects the expression of L-type voltage-gated Ca2+ channel subunit cav1.2 and receptor gated channel subunits NR1 and NR2,which may protect PC12 cells from the injury caused by ischemic/hypoxia.