In order to identify whether functional L-type calcium channels are expressed in neural stem cells(NSCs) from rat embryonic hippocampus, and whether L-type calcium channels participate in the modulation of proliferation and differentiation of NSCs, the rat embryonic hippocampal tissue was dispersed into a single cell suspension, and the dissociated cells were cultured in serum-free DMEM/F12 medium containing epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), N-2 and B27 supplement. Immunofluorescent labeling showed an expression of nestin-positive cells. Following five-days culture in differentiation medium, neuron-like and astrocyte-like cells were observed, which expressed ?-tubulin Ⅲ(Tuj1) and Glial fibrillary acidic protein (GFAP), respectively. Western blot analysis showed an expression of Cav1.2?1C subunits in NSCs, but no Cav1.3?1D subunits. Moreover, L-type calcium channel currents were recorded in those cells by using whole-cell patch clamp techniques. It was found that activation of L-type calcium channels promotes proliferation and differentiation to neuronal type of NSCs. The results indicated that rat embryonic hippocampal NSCs express functional L-type calcium channels, L-type calcium channels modulate proliferation and differentiation of NSCs.

Objective To systematically review the diagnostic value of microRNA(miRNA) quantitation in breast cancer .Meth-ods Literatures about miRNA and breast cancer diagnosis were selected by retrieving Medline ,Embase and Cochrane Library .Ac-cording to the inclusion and exclusion criteria ,the literatures were independently screened ,and a 2 × 2 contingency table was con-structed .Quality of literatures was assessed by quality assessment of diagnostic accuracy studies(QUADAS) .Statistical analysis was performed by employing Meta-Disc 1 .4 software and STATA 11 .0 .Results 16 studies were included ,which contained 1 303 patients and 711 control samples .There were threshold effects among these studies (the spearman′s correlation coefficient was-0 .758 ,P=0 .001) .A random effects model was used for meta-analysis .The summary sensitivity ,specificity ,positive likelihood ratio ,negative likelihood ratio ,and diagnostic odds ratio for miRNA in breast cancer diagnosis were 0 .77(95% CI:0 .75 -0 .79) , 0 .77(95% CI:0 .74-0 .80) ,4 .19(95% CI:2 .79-6 .30) ,0 .25(95% CI:0 .19-0 .35) ,19 .91(95% CI:9 .68-40 .95) .The area un-der curve of SROC was 0 .895 0 .Conclusion These results suggest that miRNAs have potential value to diagnose breast cancer . However ,effective diagnosis of breast cancer still needs to be conducted with assistance of clinical findings and traditional lab inves-tigations .

Objective To characterize the relationship between such phenotypes and the patterns of genetic mutations in the corresponding resistance genes in drug-resistant Mycobacterium tuberculosis (MTB)isolates in Shenzhen of China during 2007-2008.Methods According to standard of WHO,International Union Against Tuberculosis and Lung Disease(IUATLD),136 strains of MTB were collected by performing drug sensitivity test(DST)to isoniazid,rifampicin,streptomycin,ofloxacin and kanamycin on Lowenstein-Jensen in 1%proportion method.Genetic mutations in the corresponding resistance genes (rpoB,katG,rpsL,rrs1,gyrAB,rrs2)in these MTB isolates were identified by PCR,followed by DNA sequencing of the purified PCR products.The minimal inhibitory concentration(MIC)values of the aforementioned anti-tuberculosis drugs were determined for these MTB clinical isolates by two-fold dilution method in vitro.Results A total of 123 isolates were collected,73 isolates were drug resistant.50 isolates were drug susceptible.Among the isolates that were resistant to isoniazid,rifampicin,streptomycin.ofloxacin and kanamycin,the proportion of isolates that harboured mutations in the respective genes was 84.6%,93.6%,65.9%,100%,61.1%.For katG gene,the mutation detected were S315T or S315N.For rpoB,the most frequently found changes were S531L(30/44,68.2%)and H526D(9/44,20.5%)or H526R(1/44,2.3%).The reported mutations that K43R and KS8Q were founded in the rpsL locus and 491C→T and 513A→C were founded in the rrs1 gene related with streptomycin-resistant strains.For gyrA,all gyrA mutations were clustered in codons 90,91,and 94 apart from the S95T that was natural polymorphism.accounted for 81.1% of the ofloxacin-resistant isolates,and condon 91 was the most frequently mutated.No mutation were found in gyrB.The most frequent substitution were 1400 A→G(9/11,81.8%)and 1483 G→T(2/11,18.2%)in a specific region of the rrs2 gene related with kanamycin-resistant strains.No mutations except S95T of gyrA detected in the drug-susceptible isolates.The MIC values of clinical drug-resistant strains that the same drug-resistant group contains a different resistance phenotype are basically the same with the relevant resistance genes in the same mutation.Associated resistance mutations in different sites varied significantly with their MIC values.Conclusion The mutation characterization of drug-resistant and drug-suscep-tible isolates of MTB have been shown to vary according to geographic region,phenotypic characteristics exist difference in resistance levels due to different muntants of drug-resistant gene.

Acute-On-Chronic Liver Failure ; blood ; pathology ; Case-Control Studies ; Cytochrome Reductases ; metabolism ; Humans ; Prospective Studies ; alpha-Fetoproteins ; metabolism

Objective To explore the relative expression of plasma miR-199a-5p and miR-200c-3p in gastric adenocarcinoma cancer(GAC) patients and its clinical value.Methods Case-control study was used in this research.The relative expression of plasma miR-199a-5p and miR-200c-3p from 47 GAC patients and 50 healthy controls were determined by RT-PCR ( TaqMan Probe method).Meanwhile, the association with age, gender, tumor location, size, degree of differentiation, TNM stage, lymph node metastasis and other clinical pathological parameters were analyzed.The expression of these two miRNAs in plasma of 30 GAC patients during preoperation was compared with their expression 6-8 days after radical surgery.The sensitivity and specificity of plasma miRNAs expression for the diagnosis of GAC were analyzed using the receiver operating characteristic ( ROC ) curve.SPSS20.0 statistical software was used for statistical analysis.T-test, paired t-test and one-factor ANOVA were used for normal distribution of quantitative data.Results The plasma level of miR-199a-5p in GAC patients was significantly lower(1.05 ±0.22) (t =3.058,P =0.003), while miR-200c-3p was significantly higher(15.15 ±3.02) (t =-2.854,P=0.006), when they were compared with those in controls(26.80 ±8.38, 3.39 ±0.87).Low miR-199a-5p expression in GAC patients were associated with lymph node metastasis ( F =4.725, P =0.029) and the differentiation degree of gastric cancer(F=3.854,P=0.032).The relative expression of miR-199a-5p in postoperative plasma was significantly increased(t=-3.814,P=0.001), but the relative expression of miR-200c-3p was significantly reduced when compared to the preoperative samples(t=2.978, P=0.006).Area under the ROC curve of miR-199a-5p, miR-200c-3p and combined miR-199a-5p and miR-200c-3p were 0.692, 0.792 and 0.798, the sensitivity and specificity were 87%,97%,92.5% and 43%,54%, 65%, respectively.Conclusion Combined detection of miR-199a-5p and miR-200c-3p in plasma has a higher sensitivity and specificity than the conventional tumor marker CEA and CA19-9, and may be a useful combination for gastric cancer diagnosis.

Objective To preliminarily investigate the protective effect of chronic clonorchis sinesis(Cs) infestation against sepsis in Sprague Dawley(SD) rats in order to explore its underlying mechanism.Methods Chronic Cs infestation model of SD rats was reproduced by intra-gastric administration with Cs ova.Twenty rats were randomly(random number) divided into normal group(n=10) and Cs group(n=10).The proportion of differentiation in M1 and M2 macrophages were detected by flow cytometry.The expressions of Arg-1(arginine-1),FIZZ 1,iNOs and TNF-αmRNA were examined by reverse transcriptase polymerase chain reaction(RT-PCR).The cecal ligation and puncture(CLP) procedure was performed to reproduce sepsis model of SD rats.Sixty rats were randomly(random number) divided into control group,SHAM group,CLP group,Mφ+CLP group,Cs-Mφ+CLP group,and Cs-CLP group.The cumulative mortalities were calculated.The pathological changes of the lung tissue in different groups were demonstrated by HE staining.The serum levels of cytokines TNF-α and IL-10 were detected by ELISA at 0,24,48 and 72 h after CLP procedure.Results Compared with M1 peritoneal macrophages differentiation in control group(91.9%),rat peritoneal macrophages were activated to M2 differentiation(95.1%) in chronic Cs infection group.RT-PCR assay showed expression of Arg-1 and FIZZ 1 mRNA were higher in M2 macrophages,and on the contrary, the expression of iNOS mRNA expression was higher in M1 macrophages.The expression of TNF-α mRNA in M1 was significantly higher than that in M2, whereas the expression of IL-10 mRNA in M2 was higher than that in M1.The cumulative mortality of septic rats 72 h after CLP procedure were much lower in both chronic Cs infestation group and M2 macrophages adoptive transfer group(CLP group 70%vs.Mφ+CLP group 50%vs.Cs-Mφ+CLP group 30%vs.Cs-CLP group 0%,P<0.05).In these two groups,the pathological damages in lung tissues were significantly improved.The serum level of TNF-α was decreased and the anti-inflammatory IL-10 level was increased significantly in these two groups with Cs compared with other groups.Conclusion M2 macrophages polarization induced by chronic Cs infestation with M2 phenotype gene and expression of anti-inflammatory cytokine gene play key role in increasing anti-inflammatory cytokines and decreasing pro-inflammatory cytokines to allerviate organ damage and ameliorating the survival rate in septic rats.

Objective This retrospective study is designed to analyze the cardiovascular events of CTO to provide new information on secondary prevention of CHD in patients after hospital discharge. Methods 272 patients with definite diagnosis of CTO were enrolled in this study. Patients were divided into two groups according to whether suffering from cardiovascular events, with 167 patients in group A who had not suffered from the cardiovascular events and 105 patients in group B who had suffered from the cardiovascular events. We com-pared the clinical data, severity of coronary artery lesion, treatment in two groups. Results Between two groups, there was statistic signifi-cance in LDL -C, EF, Gensini scots, the number of coronary artery lesion, the number of chronic total coronary occlusion and PCI success. Logistic regression analysis revealed that Gensini scots was the independent factors for prognosis of CTO. Conclusion Gensini scors was the independent factor for prognosis of CTO.

Objective The frequency and the anatomic distribution of involved regional nodes in recurrent and locally advanced breast cancer were analyzed, in order to evaluate the rational of conventional regional node radiation technique and provide evidence for target definition of breast cancer . Methods Patients with recurrent or locally advanced breast cancer who were treated in our hospital from August 2003 to December 2009 were included in this study. 111 patients had contrast enhanced chest CT images of the whole regional nodes before treatment. The regional nodes were categorized into 8 anatomical substructures including medial and lateral supraclavicular nodes ( SC-M, SC-L), axilla nodes ( ALN )- Ⅰ , Ⅱ , Ⅲ,infraclavicular nodes (IFN), Rotter's nodes (RN) and internal mammary nodes (IMN). The frequency of involvement and anatomical distribution of the involved nodes on CT images were analyzed. Results A total of 111 patients were enrolled this study and 199 anatomical substructures with involved nodes were identified. The frequency of involvement were :SC-M 33, SC-L 21, ALN- Ⅰ 30, ALN-Ⅱ 25, ALN-Ⅲ + IFN 35, RN 27, IMN 28. Supraclavicular region and axilla were the most frequently involved area (72. 3% ).The average depth of the SC-M and SC-L nodes was 33.48 mm ± 10. 57 mm and 45.62 mm ±20. 45 mm,and 51.5% and 71.4% of the SC-M and SC-L nodes were located more than 3 cm deep from the skin. The axilla nodes were located cranial and caudal to the axillary vein in 5 and 20 locally advanced breast cancer patients and in 64 and 28 patients who received prior axillary dissection. The majority of involved IMN was located within the first 3 intercostal spaces (26/28). The average distance between the center of involved IMN and chest skin was 24. 23 mm ± 10. 28 mm. The average distance between the center of involved IMN and midline of the body was 29. 38 mm ±6. 7 mm. The center of involved IMN was 6.19 mm ±5.73 mm lateral and 5.73 mm ± 4. 56 mm posterior to the internal mammary vessels. Conclusions Conventional field design is unlikely to provide sufficient dose to the entire risk region because of individual differences.Individualized treatment planning based on CT would become feasible with increasing knowledge of natural risk of nodal involvement.

Objective To assess set-up errors measured with kilovoltage cone-beam CT (KV-CBCT), and the impact of online corrections on margins required to account for set-up variability during IMRT for patients with prostate cancer. Methods Seven patients with prostate cancer undergoing IMRT were enrolled onto the study. The KV-CBCT scans were acquired at least twice weekly. After initial set-up using the skin marks, a CBCT scan was acquired and registered with the planning CT to determine the setup errors using an auto grey-scale registration software. Corrections would be made by moving the table if the setup errors were considered clinically significant ( i. e. , ＞ 2 mm). A second CBCT scan was acquired immediately after the corrections to evaluate the residual error. PTV margins were derived to account for the measured set-up errors and residual errors determined for this group of patients. Results 197 KV-CBCT images in total were acquired. The random and systematic positioning errors and calculated PTV margins without correction in mm were:a) Lateral 3. 1,2. 1,9. 3;b) Longitudinal 1.5, 1.8, 5. 1 ;c) Vertical 4. 2,3.7, 13.0. The random and systematic positioning errors and calculated PTV margin with correction in mm were:a) Lateral 1.1,0. 9, 3.4;b) Longitudinal 0. 7, 1.1, 2. 5;c) Vertical 1.1, 1.3, 3.7. Conclusions With the guidance of online KV-CBCT, set-up errors could be reduced significantly for patients with prostate cancer receiving IMRT. The margin required after online CBCT correction for the patients enrolled in the study would be appoximatively 3-4 mm.

OBJECTIVETo investigate the clinical features, surgical transcranial approaches and outcomes of giant pituitary adenomas.

METHODSA series of 112 consecutive cases of giant pituitary adenomas underwent microsurgery through transcranial approaches at People' s Liberation Army General Hospital were retrospectively analyzed. Of the 112 patients, 58 were male and 54 were female, with age ranging from 3 to 72 years(mean age 44. 3 years). There were 91 non-functioning adenomas and 21 hormone-secreting adenomas. The maximum tumor diameter varied from 4. 0 to 7. 2 cm, with mean diameter of 4. 8 cm. Unilateral subfrontal approach was chosen in 16 cases, pterional approach in 41 cases, anterior interhemispheric approach in 34 cases, transcallosal-interforniceal approach in 6 cases, transcortical- transventricular approach in 5 cases, combined approach in 6 cases and other approaches in 4 cases. Postoperative MRI and endocrine function were re-examined routinely to evaluate the therapeutic efficacy. Staged operation through transsphenoidal approach or adjuvant treatments including medical and radiation therapies were administered in patients with hormone-secreting adenomas when hormonal excess persisted after surgery and in patients with non-functioning adenomas who had postoperative MRI evidence of residual tumor.

RESULTSTotal removal of the lesion was achieved in 57 cases (50. 9%) , 26 patients (23. 2%) underwent subtotal resection, and 29 patients (25. 9%) underwent partial removal. Postoperative mortality occurred in 3 patients (2. 7%). Major surgical morbidity occurred in 38 patients (33. 9%). Vision was preserved or improved in 98 patients (87. 5%). The postsurgical follow-up period varied from 3 to 64 months (mean 19. 5 months) . Nineteen of the 21 patients with hormone-secreting adenomas were considered to be in hormonal remission, and 87. 2% of the cases were capable of normal work and life and 8 patients suffered recurrence during the followed-up period.

CONCLUSIONSSelection of appropriate transcranial approaches is the key to successful microsurgery for giant pituitary adenomas according to the morphological characteristics presented in image examinations and clinical symptoms. Staged transsphenoidal operation and/or adjuvant therapies including medical and radiation therapies offer the best chances to control the residual tumors after the maximal surgical removal of giant adenomas through transcranial approaches.

Adenoma ; surgery ; Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; Combined Modality Therapy ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Microsurgery ; methods ; Middle Aged ; Neurosurgical Procedures ; methods ; Pituitary Neoplasms ; surgery ; Postoperative Period ; Retrospective Studies ; Young Adult