Objective To explore efficacy and mechanism of recombinant human granulocyte-colony stimulating factor(rhG-CSF)in treatment of different leukopenia.Methods A total of 50 patients were divided into 4 groups:aplastic anemia(AA)group(n=10),myelodysplastic syndromes(MDS)group(n=10),drug-induced leukopenia group(n=20)(subdivided into hyperplasia group and hypoplasia group by the degree of cellularity)and iron deficiency anemia group(control group,n=10).The concentrations of G-CSF of peripheral blood were measured by ELISA method.The ratio of CD34+ cells and G-CSFR expression in blood marrow mononuclear cells(BMMNC)were measured by flow cytometry.Granulocyte colony-forming units(CFU-G)of MNC were cultured.Clinical efficacy of rhG-CSF to all patients were determined.Results The concentrations of G-CSF were higher in AA and drug-induced leukopenia groups than those of MDS and control groups(P0.05).Expressions of G-CSFR were lower in hypoplasia group than those in hyperplasia and control groups(P0.05).The ratio of CD34+ cells was lower in AA group than that of other groups(P0.05).Concentrations of CFU-G were lower in AA and MDS groups than those in drug-induced leukopenia and control groups(P0.05).The efficacy of rhG-CSF was superior in drug-induced leukopenia group than in MDS group,and superior in MDS group than in AA group(P

Objective To assess whether dynamic arterial elastance (Eadyn)can be used to predict the reduction of arterial pressure after decreasing norepinephrine (NE) dosage in patients with septic shock.Methods A prospective observational cohort study was conducted.Thirty-two patients with septic shock and mechanical ventilationwere enrolledfrom January 2014 to December 2015 in ICU of Wuxi People's Hospital of Nanjing Medical University.Hemodynamic parameters were recorded by pulse contour cardiac output(PiCCO) monitoring technology before and after decreasing norepinephrine dosage.Eadyn was defined as the ratio of pulse pressure variation (PPV) to stroke volume variation (SVV).Mean arterial pressure (MAP) variation was calculated after decreasing the dose of NE.Response was defined as a ≥ 15％decrease of MAP.AUC was plotted to assess the value of Eadyn in predicting MAP response.Results A total of 32 patients were enrolled in our study,with 13 responding to NE dose decrease where as the other 19 did not.Eadyn was lower in responders than in nonresponders (0.77 ± 0.13 vs 1.09 ± 0.31,P ＜ 0.05).Baseline Eadyn was positively correlated with systolic blood pressure variation,diastolic blood pressure variation,systemic vascular resistance variation and MAP variation (r =0.621,P =0.000;r =0.735,P =0.000;r =0.756,P =0.000;r =0.568,P =0.000 respectively).However,stoke volume variation,baseline level of systemic vascular resistance and NE baseline dose were not correlated with Eadyn baseline value (r =0.264,P =0.076;r =0.078,P =0.545;r =0.002,P =0.987 respectively).Eadyn ≤ 0.97 predicted a decrease of MAP when decreasing NE dose,with an area under the receiver-operating characteristic curve of 0.85.The sensitivity was 100.0％ and specificity was 73.7％.Conclusions In septic shock patients treated with NE,Eadyn is an index to predict the decrease of arterial pressure in response to NE dose reduction.

Objective To observe the effects of polysaccharide extracted from Yangwei Mixture on immunologic functions in mice.Method Immuno-suppressant mice model was established by intraperitoneal injection of CTX.The effects of the polysaccharide were observed through phagocytosis test of mice celiac macrophagocyte and serum hemolysin level test as well as lymphocyte transformation test.Result The polysaccharide can obviously increase the percentage of phagocytosis of mice celiac macrophagocyte,accelerate the formation of hemolysin and promote the lymphocyte transformation.Conclusion Polysaccharide extracted from Yangwei Mixture is a good immuno-stimulant,and can counteract the immuno-suppression induced by CTX.

Purpose To explore the expression of Aurora kinase A (Aurora-A), minichromosome maintenance protein 7 (MCM7) and human papillomavirus type 16 E7 protein (HPV 16 E7) in uterine cervical squamous cell carcinoma (CSCC) and to investigate their relationship with clinicopathological factors. Methods Immunohistochemical method was employed on 20 cases of low-grade cervical intraepithelial neoplasia (CIN1) , 30 cases of high-grade cervical intraepithelial neoplasia (CIN2+3), 40 cases of CSCC, and 20 ca-ses of chronic cervicitis. Results (1) Aurora-A localized in the cytoplasm and nucleus. MCM7 protein positive staining localized in the nucleus. In the nucleus, and (or) the cytoplasm appeared brown particles positive for HPV 16 E7. (2) The expression of Aurora-A, MCM7 and HPV 16 E7 were higher in the group of CIN2+3 and CSCC than that in the group of chronic cervicitis or CIN1 ( P<0. 0083). (3) In cervical cancer, the expression of Aurora-A, HPV 16 E7 showed positive correlation (P<0. 001, rs =0. 657). The expression of MCM7, HPV 16 E7 showed positive correlation (P<0. 001, rs =0. 616). The expression of Aurora-A, MCM7 showed positive correlation (P<0. 001, rs =0. 597). (4) Aurora-A expression levels was associated with tumor cell differentiation, clinical stage and lymph node metastasis (P<0. 05). MCM7 expression levels was associated with tumor cell differentiation and clinical stage (P<0. 05). HPV 16 E7 expression had no correlation with patient age, tumor size, tumor differentiation, clinical stage and lymph node metastasis (P>0. 05). Conclusion Aurora-A, MCM7 and HPV 16 E7 expression are gradually increased with disease progres-sion, and closely related to the occurrence and development of cervical cancer, they are expected to be early diagnosis, early treatment of biological indicators of cervical cancer.

Objective: To investigate the effects of taurine on blood glucose, number of platelets in plasma, pletelet aggregation and metabolism of oxygen free radicals in diabetic rats. Methods: Diabetes was induced by a single injection (ip) of streptozotocin (STZ,70 mg/kg body weight). The rats were maintained on drinking water with or without taurine (1.5 g/kg body weight per day). The concentration of blood glucose, maximal aggregation rate of platelets induced by adenosine 5 diphosphate(ADP) or thrombin(Thr) and the number of platelets in plasma were measured. Metabolism of oxygen free radicals in platelets was assessed as superoxide dismutase(SOD) activity and content of malondialdehyde(MDA) in platelets. [WT5FZ]Results: [WT5BZ]The concentration of blood glucose and maximal aggregation rate of platelets induced by ADP or Thr were significantly higher in diabetic rats than in non diabetic rats. The diabetic rats showed decreased platelet number and SOD activity with higher MDA level. In taurine treated diabetic rats concentration of blood glucose was obviously decreased, but no significant change was found in platelet number, SOD activity, MDA content and aggregation of platelets induced by ADP or Thr. [WT5FZ] Conclusion: [WT5BZ]There is a correlation between the increase in platelet aggregability and the abnormal metabolism of oxygen free radicals in platelets and taurine can decrease the blood glucose level but has no significant effect on platelet aggregability and metabolism of oxygen free radicals in diabetic rats.

Soluble receptor for advanced glycation end products (sRAGE) can decoy the toxic AGEs and is considered to be a protective factor.This study aimed to evaluate the correlation between intrafollicular sRAGE levels and clinical outcomes in infertile women of young or advanced maternal age (AMA) undergoing in vitro fertilization (IVF).A total of 62 young women and 62 AMA women who would undergo IVF were included in this prospective study.The intrafollicular sRAGE concentration was measured to determine its association with the number of retrieved oocytes,fertilized oocytes,high-quality embryos or achievement of clinical pregnancy in young and AMA women,respectively.Besides,correlations between sRAGE and age or follicle-stimulating hormone (FSH) were examined.We found that the intrafollicular sRAGE levels were higher in young patients than those in AMA patients,suggesting that the sRAGE levels were inversely correlated with age.In young patients,sRAGE showed no correlation with the number of retrieved oocytes,fertilized oocytes,high-quality embryos or achievement of clinical pregnancy.But it was found that AMA patients with more retrieved oocytes,fertilized oocytes and high-quality embryos demonstrated higher sRAGE levels,which were a prognostic factor for getting clinical pregnancy independent of age or FSH level.In conclusion,the sRAGE levels decrease with age.Elevated intrafollicular sRAGE levels indicate good follicular growth,fertilization and embryonic development,and successful clinical pregnancy in AMA women,while in young women,the role of sRAGE may not be so predominant.

The development of TCM acupuncture represents a internationalized and modern trend. A recent study with the title of "Acupuncture for chronic knee pain: a randomized clinical trial" published in Journal of the American Medical Association on October 1st, 2014, which raised doubts on acupuncture efficacy as well as traditional manipulation and acupoint theory, makes some negative impact and challenges on the development of acupuncture. From the view of future development of acupuncture, the potential influence of this research on acupuncture development is proposed, and by combining acupuncture theory, some discussions and doubts on the research design and outcome explanations are made. Additionally, enlightenments of this research on further clinical research are summarized.
Acupuncture Therapy ; Chronic Pain ; therapy ; Female ; Humans ; Low-Level Light Therapy ; Male

BACKGROUND: Schizophrenia is substantially heritable, but specific susceptibility genes remain difficult to be identified. Therefore, it is necessary to explore hereditary markers first.OBJECTIVE: To investigate the relationship between schizophrenia and related vWA allele genes based on the analysis of microsatellite DNA vWA polymorphism.DESIGN: A case-controlled study with schizophrenic patients and randomly selected population as subjects.SETTING: Ward of Dalian Seventh People's Hospital and Molecular Biological Laboratory of Dalian Medical University.between March and July 2002 at Dalian Seventh People's Hospital which specializes in schizophrenia. Schizophrenia was diagnosed according to the diagnostic standard of the third edition of "the American Diagnostic Statistical Manual for Schizophrenic Diseases", and their clinical manifestations were predominantly negative signs. Altogether 123 normal blood samples were collected from random population at the Blood Center of Dalian Red Cross. They all denied psychological ailments and severe systematic diseases, and they had no kinship with each other.METHODS: Heparin anti-coagulation blood samples were collected and PCR compound amplification was carried out with the aid of PE Profiler plus system. Then the products were subjected to electrophoresis and gene detection with ABI310 type gene analysis system so as to calculate the frequency of allele genes; Hardy-Weinberg equation law was used to make coincidence test and linkage analysis of the theoretical frequency and actual one. Schizophrenic patients and random population were compared and relative risk was calculated with RR=Pd × (1-Pc)/Pc × (1-Pd) in order to assess the statistical significance (RR: relative risk; Pd: gene frequency of schizophrenia; Pc: gene frequency of random population). RR ＞ 1 was considered of higher susceptibility while RR ＜ 1 was considered of anti-susceptibility. In this way, we could find out vWA allele genes that had susceptible linkage or anti-linkage with schizophrenic related genes.MAIN OUTCOME MEASURES: Major outcome: Correlation analysis of vWA allele genes in schizophrenic patients and random population. Secondary outcome: The coincidence of vWA allele gene frequency in patients with schizophrenia and random population with what was calculated by Hardy-Weinberg law.RESULTS: Data of the two groups were processed according to the objective and statistically analyzed.① vWA allele gene frequency in patients with schizophrenia and in random population was found to coincide with HardyWeinberg law(P ＞ 0.05).② The positive rate of vWA-14 in schizophrenic patients (17.2%) was obviously different from that in random population (33.3%) (RR=0.415, P=0.014). The positive rate of vWA-17 in schizophrenic patients (31.3%) was found to be significantly higher than that in random population (19.5%) (RR=1.866, P=0.043) while it did not differ significantly in other allele genes (P ＞ 0.05).CONCLUSION: The positive rate of vWA-14 was significantly lower in schizophrenic patients than in random population, indicating that vWA-14locus may be negatively selected in schizophrenia due to some reasons,which may be approximate to anti-schizophrenia genes. Moreover, the higher expression of vWA-17 in schizophrenic patients than in random population suggests that vWA-17 locus is correlated with schizophrenia,which may be approximate to schizophrenia-susceptibile genes.

Objective To explore the effects of under-expression of large tumor suppressor 1 (LATS1) on activation of Hippo signaling pathway and differentiation, proliferation, migration of bone marrow mesenchymal stem cells (mMSCs) of micein vitro.Methods mMSCs of C57BL/6 mice were divided into normal control (MSC) group, empty vector control (MSC-GFP) group, LATS1-over-expressing (MSC-LATS1) group, empty vector without LATS1 shRNA control (MSC-shControl) group and LATS1-under-expressing (MSC-shLATS1) group. Lentiviral vectors with activated,inactivated LATS1 (the key molecule of Hippo signaling pathway) modifications and empty vectors were constructed and were used to infect mMSCsin vitro. The transduction efficiencies mediated by the lentiviral vectors were evaluated by fluorescence microscopy and flow cytometry. The mRNA expression of LATS1 was quantified by quantitative real-time polymerase chain reaction (qRT-PCR), and the protein expressions of LATS1, YAP (p-YAP), 14-3-3 were quantified by Western Blot to evaluate the activation of Hippo signaling pathway. Osteogenic and adipogenic differentiation of mMSCs were evaluated through measurement of Runx2, OSX and C/EBPα, PPAR-γ mRNA by qRT-PCR, as well as Alizarin Red S and Oil red O staining. Proliferation of mMSCs was evaluated using methy thiazdyl tetrazolium (MTT) assay. The scratch test and Transwell chamber test were used to analyze the horizontal and vertical migration ability of mMSCs.Results The transduction efficiencies mediated by the lentiviral vectors were 94.74%-96.10%. Compared with MSC-GFP group, the activation of Hippo signaling pathway was promoted in MSC-LATS1 group [LATS1 mRNA (2-ΔΔCT): 4.37±0.21 vs. 1.20±0.04, LATS1 protein (gray value): 2.21±0.06 vs. 1.09±0.10, p-YAP/YAP protein (gray value): 1.51±0.13 vs. 0.98±0.05, 14-3-3 protein (gray value): 1.92±0.18 vs. 1.10±0.09, allP < 0.05], osteogenic and adipogenic differentiation of mMSCs were decreased in MSC-LATS1 group [mineralization (A value):0.13±0.02 vs. 0.40±0.03, Runx2 mRNA (2-ΔΔCT): 0.51±0.02 vs. 0.98±0.09, OSX mRNA (2-ΔΔCT): 0.41±0.04 vs. 1.04±0.09, lipid accumulation (A value): 0.10±0.02 vs. 0.25±0.03, C/EBPα mRNA (2-ΔΔCT): 0.33±0.03 vs. 1.11±0.09, PPAR-γ mRNA (2-ΔΔCT): 0.29±0.02 vs. 1.04±0.10, allP < 0.05], the proliferation rate of mMSCs at 4-7 days was decreased in MSC-LATS1 group and so were the horizontal and vertical migration of mMSCs [wound healing rate: (18.65±3.53)% vs. (40.29±1.87)%, migrated cells (cells/MP): 35.99±6.18 vs. 103.67±17.77, bothP <0.05]. Compared with MSC-shControl group, the activation of Hippo signaling pathway was inhibited in MSC-shLATS1 group [LATS1 mRNA (2-ΔΔCT): 0.16±0.01 vs. 0.98±0.03, LATS1 protein (gray value): 0.38±0.03 vs. 1.04±0.07, p-YAP/YAP protein (gray value): 0.58±0.04 vs. 1.05±0.06, 14-3-3 protein (gray value): 0.14±0.02 vs. 1.02±0.09, allP < 0.05], osteogenic and adipogenic differentiation of mMSCs were increased in MSC-shLATS1 group [mineralization (A value): 0.93±0.13 vs. 0.44±0.05, Runx2 mRNA (2-ΔΔCT): 1.44±0.12 vs. 0.95±0.04, OSX mRNA (2-ΔΔCT):1.67±0.06 vs. 1.10±0.11, lipid accumulation (A value): 0.47±0.06 vs. 0.28±0.04, C/EBPα mRNA (2-ΔΔCT):3.98±0.61 vs. 0.99±0.10, PPAR-γ mRNA (2-ΔΔCT): 3.05±0.36 vs. 0.98±0.14, allP < 0.05], the proliferation rate of mMSCs at 3-7 days was increased in MSC-shLATS1 group and so were the horizontal and vertical migration of mMSCs [wound healing rate: (80.18±6.98)% vs. (46.18±1.01)%, migrated cells (cells/MP): 212.69±41.21 vs. 115.87±35.15, bothP < 0.05].Conclusions Under-expression of LATS1 promotes the differentiation, proliferation, migration of mMSCs by inhibition of Hippo signaling pathwayin vitro.

Objective To establish a TaqMan real-time fluorescent quantitative PCR to detect Vibrio vulnificus based on hemolysin gene(vvhA)that coding cytolysin.Method By using software Primer Express, the PCR primers and TaqMan probe,which located in the conserved region of vvhA gene sequence,were designed for establishment of a TaqMan real-time fluorescent quantitative PCR to detect 100 bp amplicon from V.vulnificus DNA.A recombinant plasmid pMD19-vvhA100 as a positive control during detection was constructed using gene cloning technique.Minimal amplification cycles(Ct value)and fluorescence intensity enhancement (△Rn value)were used as observing index to optimize the reaction conditions of the TaqMan real-time fluorescent quantitative PCR.The DNAs with different concentrations from V.vulnificus and other eight bacteria and pMD19- vvhA100 were applied as templates to determine the specificity,sensitivity and reappearance of the TaqMan real- time fluorescent quantitative PCR.ICR mice were intraperitoneally,subcutaneously and orally infected with V. vulnificus,respectively.The detection effect of the TaqMan real-time fluorescent quantitative PCR was measured using the specimens of peripheral blood,subcutaneous tissue and intestinal content collected from the infected mice.Results The established TaqMan real-time fluorescent quantitative PCR showed positive results only for V. vulnificus DNA and pMD19-vvhA100.The detection effectiveness of the TaqMan real-time fluorescent quantitative PCR was as high as 0.01 ng of V.vulnificus DNA or 103 copies of pMD19-vvhA100.The SD values of the detection results repeated for three times using pMD19-vvhA100 with different concentrations were lease than 0.79. The detection results of TaqMan real-time fluorescent quantitative PCR were positive for all the specimens of peripheral blood and subcutaneous tissue.Conclusions The TaqMan real-time fluorescent quantitative PCR established in this study for V.vulnificus vvhA gene detection has advantages such as quickness,stability, sensitivity and specificity,indicating this method can be used for clinical laboratory diagnosis of septicemia and wound infection caused by V.vulnificus.