Objective　To study the occurence, development and regulation of reactive gliosis with astrocyte (Ast) in vitro. Methods　Ast was isolated and cultured in vitro and its model of reactive gliosis was established by scratching the cultured astrocytes. The reactivity and rules of Ast to injury was studied by morphological changes, RT-PCR, immunocytochemistry, in situ hybridization and imaging analysis. Results　After scratching, the astrocytes showed typical features of reactive gliosis, with the hypertrophic cell body, thickened and lengtheded processes, and enhanced glial fibrillary acidic protein (GFAP) staining. In situ hybridization and RT-PCR analysis confirmed that the expression of GFAP mRNA was markedly increased. These changes occurred 1 d after scratching and reached the peak 5 to 7 d after injuring. Conclusion　A model of reactive astrogliosis was successfully established in vitro which showed an active reaction to injury. The characteristics of reactive gliosis parallel that seen in vivo.

Objective To analyze distribution and antibiotic resistance pathogenic bacteria in neonatal sepsis in 10 years. Methods The distribution of pathogens and their antibiotic resistance were retrospectively analyzed in neonatal sepsis from January, 2004 to December, 2013. The results were compared between 2004-2008 and 2009-2013. Results The percentage of Streptococcus agalactiae rose from 1.0%to 4.2%and fungi rose from 1.9%to 7.9%in all pathogens in past 10 years. But the distribution of pathogenic bacteria was not significantly different. The ESBLs of Escherichia coli were increased from 28.6%to 36.0%(P>0.05). The Escherichia coli resistant to imipenem and meropenem had not been found. The Staphylococcus resistance to oxacallin and ampicillin/sulbactam was increased. The Staphylococcus resistant to vancomycin had not been found. Conclusions The main pathogens of neonatal sepsis are coagulase-negative staphylococci and E. coli. The fungi and Streptococcus agalactiae infections are signiifcantly increased.

Objective To investigate the effects of mutated PS-1 on the proliferation and apoptosis of RA-induced PC12 cells . Methods The MTT assay, flow cytometry, TUNEL method were used in the research. Results An inhibition of proliferation was demonstrated in the RA-induuced PC12 cells expressing PS-1 mutant L286V. The proliferation of the cells was blocked in the G1→S phase. Whether with or without fetal bovine serum(FBS), PS-1 mutant L286V was able to accelerate the apoptosis of RA-induced PC12 cells. This action was enhanced markedly under the culture without FBS. Conclusions The RA-induced PC12 cells expressing PS-1 mutant L286V shows that the proliferation is inhibited and blocked in the G1→S phase. Whether with or without FBS, PS-1 mutant L286V can accelerate the apoptosis of RA-induced PC12 cells. This action is enhanced markedly under the culture without FBS.

Objective To investigate the cholinergic neuron properties and the changes of proliferation and apoptosis of PC 12 cell induced by different concentrations of all trans retinoic acid(RA). Methods PC 12 cell was induced by RA of concentrations of 1-50 ?mol/L. Changes of cell growth curve and cycle, apoptosis and changes of activities of acetylcholinesterase(AchE) and choline acyltransferase(ChAT) were detected by MTT assay, flow cytometry, TUNEL method, spectrophotometry and radiochemistry assay, respectively. Results ① RA significantly enhanced the activity of ChAT with concentrations between 1 to 20 ?mol/L. The activity of ChAT with concentration of 10 ?mol/L was higher than those with other concentrations. ② The apoptosis of RA induced PC 12 cells was markedly enhanced when concentration was higher than 10 ?mol/L. ③ A concentration dependent inhibition of proliferation was demonstrated in the RA induced PC 12 cells. ④ PC 12 cells proliferation was blocked in the G 1→S phase. Conclusion RA can be used to induce PC 12 cells to show the properties of cholinergic neuron with an optimal concentration of 10 ?mol/L.

Objective To study the relationship between GFAP and the malignant degree of astrocytoma. Methods After successful transfer of retrovirus with GFAP (PLBsKG) into C6 cells mediated with liposome, the morphology, growth curve and the proliferative cycle of the target cells were observed. The expression of the protein and mRNA of GFAP was determined with RT-PCR and immunocytochemical staining. In addition, the relationship between the staining GFAP-IR and the malignant degree of astrocytoma was analyzed according to Kermohan′s grading standard in 20 samples of the cells of astrocytoma with imaging analytic system. Results PLBskG resulted in the decrease of GFAP in the target C6 cells, and changed the morphology and increased the proliferation of C6 cells. The number of the cells in the G 2+M phages was increased. The intensity of GFAP-IR staining was negatively correlated to the malignant potency of astrocytoma. The more intensive staining of the cells, the lower of the malignant degree of astrocytoma and vice versa. Conclusion There is a close relationship between GFAP and malignant degree of astrocytoma. The level of GFAP expression serves the index of the malignant degree and prognosis of astrocytoma in certain degree.

Objective To explore the value of application of choledochofiberscopy in the diagnosis and treatment of the extrahepatic bile duct disease, and the effect on reducing the incidence of the postoperative residual stone in biliary ducts. Methods According to the case history and ultrasonography,if the common bile duct(CBD) diseases suspected,the CBD was explored by intraoperative choledochofiberscope(IOCF). During the procedure,a biliary passage mirror inducer apparatus and biliary tract probe which were manufactured by ourselves were used. Results During LC,IOCF was performed on 385 cases of the 10 396 LC cases,and possitive findings were dicovered in 102 cases(26.49%). Among those positive patients, 67 cases belonged to stricture of the lower biliary tract; 5 cases were Mirizzi syndrome; 2 cases were carcinoma of the periampulla; 1 case was primarily carcinoma of the bile duct; 1case was ascarisis of the biliary system. Conclusions IOCF is a good inspect technique with high success rate and clear image of bile duct, it can discover the common duct diseases which are difficult to be diagnosed through the routine examination.At the same time, it can provide the locative and qualitive diagnosis, determine reasonable methods of operation,and effectively provent postoperative complications.

Objective: To compare the short-term and long-term outcome between robotic gastrectomy and laparoscopic gastrectomy. Methods: The clinical data of 517 patients who had received robotic gastectomy and laparoscopic gastrectomy between December 2011 and December 2013 at Department of General Surgery, Chinese People′s Liberation Army General Hospital was collected. After propensity score matching, 70 patients in robotic gastectomy and 70 patients in laparoscopic gastectomy were identified. Perioperative outcome and overall survival were compared between the two groups using t test, χ² test, Kaplan-Meier curve and Log-rank test, respectively. Prognosis factors were analyzed by Cox′s proportional hazards regression. Results: There were comparable baseline characteristics between patients in robotic group (RG) and those in laparoscopic group (LG). The conversion rate for RG and LG were 5.7% and 4.3% respectively (P=1.000). Compared with LG, RG had similar lymph node retrieval (25.5±7.2 vs. 24.5±8.3, t=0.770, P=0.443) and less blood loss ((147.0±96.8) ml vs. (188.0±111.2) ml, t=-2.326, P=0.021). There were also similar complications (χ²=0.233, P=0.629) and severity of complications (W=70.500, P=0.053). Although there tended to be early mobility, early flatus and less hospital stay for patients in RG group, the difference between RG and LG was not statistically significant. The 3-year survival rate was 72.9% and 60.0% for patients in RG and patients in LG (P=0.578). Multivariable analysis revealed gender (HR=2.529, 95% CI: 1.042 to 6.140, P=0.040), neoadjuvant chemotherapy (HR=0.272, 95% CI: 0.104 to 0.710, P=0.008) and vascular invasion (HR=2.135, 95% CI: 1.027 to 4.438, P=0.042) were independent prognostic factors. Conclusion Compared with laparoscopic gastrectomy, robotic gastectomy could achieve similar short-term and long-term outcomes.

In this study, the current status for breast diseases in a region with high-incidence of cervical cancer were epidemiologically investigated. From March to August, 2009, 17618 women, from Wufeng area of Hubei province, China, were recruited to screen breast diseases by using breast infrared diagnostic apparatus. Other diagnostic methods, such as B-mode ultrasound, X-ray mammography, needle biopsy and pathological examination were, if necessary, used to further confirm the diagnosis. The screening showed that 5990 of 17618 cases (34.00%) had breast diseases, 5843 (33.16%) had mammary gland hyperplasia, 48 (0.27%) had breast fibroadenoma, 11 (0.06%) had breast carcinoma, and 88 (0.50%) had other breast diseases. The peak morbidity of breast cancer was found in the women aged 50-60 ages. The morbidity of breast cancer was significantly increased in women elder than or equal to 50 years old (n=8, 0.157%) in comparison with that in the subjects younger than 50 years old (n=3, 0.024%) (u=2.327, P<0.05). It was shown that the occurrence of breast diseases was concentrated in women aged 20-40 years, while the total morbidity reached its peak at the age of 30 years and then decreased sharply after age of 40. Compared with the patients elder than or equal to 40 years old (n=3289, 27.46%), the morbidity rate of breast diseases was significantly increased in women less than 40 years old (2648 cases, 47.18%; P<0.001). However, there was no significant difference in the morbidity of breast diseases between the age group of 20-29 years and that of 30-39 years (P=0.453), and both of them were high. There was no significant association between the morbidity of breast diseases and cervical cancer. Since the morbidity of breast diseases was higher among young women, more attention should be paid to the screening of breast diseases among young women for early diagnosis.

Objective To understand the genetic and epidemiologic characteristic of Brucella (B.) melitensis strains isolated in Guizhou province in 2010-2012. Methods B. genus specific BCSP31-PCR and species-specific AMOS-PCR were used to identify the bacteria strain,while the identified strains were analyzed under MLVA-16 and cluster analysis of B. melitensis strains. The strains were isolated from Guizhou and other provinces. Results Six B. melitensis strains were identified as B. melitensis using the BCSP31-PCR and AMOS-PCR. Data from the MLVA-16 analysis revealed the differences of repeated numbers at parts of the VNTR locus in the six strains isolated in Guizhou province. The six strains from Guizhou province and 105 B. melitensis strains from other province could be divided into 72 MLVA types(MT). Strain ZY and ZA from Guizhou province were typed as MT63,and LL3,LL4 and LL11 were typed as MT67,while strain SQ was typed as MT72. Data from the clustering analysis showed that ZY,ZA,LL3,LL4 and LL9 were most closely clustered with B. melitensis isolates from Yunnan,Fujian and Guangdong provinces,but strain SQ was genetically remote from other strains. Conclusion PCR methods,combined with MLVA-16, identified the six B. melitensis strains isolated in Guizhou province in 2010-2012 as B. melitensis biovar 3,with the genetic diversity of the strains showed. Six strains were closely related to the B. melitensis strains from Yunnan,Fujian and Guangdong provinces. The results of this study provided scientific basis for the control and prevention of Brucellosis in Guizhou province.

This paper summarized Professor LI Haisong's clinical experience in treating spermatorrhea based on the view of "spirit controlling essence and qi". It is emphasized that the heart spirit has the function of controlling the essence and qi of the human body， believing the frenetic stirring of heart spirit and the insecurity of essence gate are the core pathogenesis of spermatorrhea， LI advocates to regulate the heart spirit first and take into account of the zang-fu organs， essence and qi simultaneously for the treatment. Treatment should be performed according the clinical syndromes differentiatied. For those with heart spirit failing to nourish syndrome， it is recommended to supplement heart qi， nourish spirit and consolidate essence with self-made Yangxin Mijing Formula （养心秘精方）. In case of heart fire hyperactivity， the method of clearing heart heat and draining fire， calming spirit and consolidating essence should be used， and self-made Xiexin Gujing Formula （泻心固精方） is recommended. For heart-liver qi constraint， it is advised to soothe the liver and calm heart， calm the mind and regulate essence with self-made Jieyu Anshen Tiaojing Formula （解郁安神调精方） which is a modifcation to Chaihu Shugan Powder （柴胡疏肝散）. In terms of deficiency of both heart and liver， the treatment principle is supplementing spleen and nourishing heart， calming the mind and controlling essence， for which self-made Xinpi Tongtiao Shejing Formula （心脾同调摄精方） modified from Guipi Decoction （归脾汤） can be used. For deficiency of both heart and kidney， it is better to nourish the kidney and calm heart， calm the mind and consolidate essence with self-made Xinshen Liangzi Tianjing Formula （心肾两滋填精方） that modified from Shuilu Erxian Elixir （水陆二仙丹） and Wuzi Yanzong Pill （五子衍宗丸）. Prescriptions are used to treat the root by harmonizing the zang-fu organs， nourish the spirit by regulating qi and blood， and calm the mind by taking special medi-cinals， and they should be flexibly modified according to the disease.