BACKGROUND: Nerve growth factor (NGF) is characterized by poor stability both in vitro and in vivo, and liable to lose its bioactivity.OBJECTIVE: To study the stability of liposome-encapsulated NGF injection preserved under various conditions.DESIGN: A controlled study of liposome-encapsulated NGF.SETTING: Department of Surgery of the First Affiliated Hospital of Nanjing Medical University.MATERIALS: This study was carried out at the Central Laboratory of the First Affiliated Hospital of Nanjing Medical University between July 2002and March 2004. NGF from rat submaxillary gland was purified, encapsulated by liposome, and prepared into lyophilized dosage form before preserved under different conditions (at 4 ℃, room temperature, 40 ℃ or 40 ℃ with saturated humidity, respectively).METHODS: Chicken embryo dorsal root ganglion cultured in serum-free medium was used to evaluation the bioactivity of NGF in vitro. The dorsal root ganglion from 8-day-old chicken embryo was inoculated in a polylysine-coated 24-well culture plate and cultured in Dulbecco modified Ea-gle medium (DMEM) containing different testing samples. Only DMEM was used for culture in the negative control group, while DMEM containing NGF at different concentrations used in the positive control group. The ganglion was cultured at 37 ℃ with 50 mL/L CO2 and saturated humidity for 24 hours, and the growth of the nerve fibers was observed under an inverted microscope. The bioactivity of NGF was also evaluated in simulated condition in vivo by adding lyophilized liposome-encapsulated NGF and positive control NGF specimen into 0.5 mL rat serum, which, along with the blank control serum, was added into 2.5 mL DMEM at 0.5, 1, 2, 3, 4,6 hours and thoroughly mixed. The bioactivity of NGF was assessed accord-ing to the length and density of the dorsal root ganglion and graded the prominences (recorded as "++" or "+++ "), and very long and dense growth of the prominences (++++).MAIN OUTCOME MEASURES: In vitro bioactivity of NGF preserved for 10, 30, 60, and 90 days by testing the growth of cultured chick embryo dorsal root ganglion in serum-free DMEM and in test of rats serum containing NGF added at 0, 0.5, 1, 2, 3, 4, and 6 hours into DMEM.RESULTS: Lyophilized liposome-encapsulated NGF exhibited stable bioactivity (+++) after preservation at 4℃ and room temperature for 10-90days; at 40 ℃ for 10-60 days, the it retained its the bioactivity (+++),which, however, slightly decreased by 90 days (++); its bioactivity was preserved (+++) at 40 ℃ with saturated humidity for 10 days (+++), slightly decreased at 30-60 days (++) and noticeably lowered (+) at 90 days. When preserved for 0, 0.5, 1, 2, and 3 hours in rat seum, the NGF preparation retained stable bioactivity (++++ or +++), which slightly decreased at 4 hours and 6 hours (++).CONCLUSION: Liposome-encapsulated NGF has stable bioactivity but its preservation at relatively high temperature with high humidity is difficult.Lyophilized liposome-encapsulated NGF exhibits better bioactivity than NGF-liposome suspension after preservation under various conditions.

Objective To investigate the occurrence of postoperative healthcare-associated infection(HAI)and its risk factors in neurosurgical patients undergoing removal of intracranial tumor,so as to provide theoretical basis for formulating intervention measures.Methods Prospective survey was adopted to monitor the occurrence of postoperative HAI in patients who admitted to the department of neurosurgery of a hospital and underwent selective removal of intracranial tumor between April 2013 and December 2014 ,risk factors for HAI were analyzed with univariate and multivariate logistic regression analysis.Results A total of 1 218 patients were surveyed,163 patents developed 193 times of postoperative HAI,inci-dence of postoperative HAI was 13.38%,case incidence of HAI was 15.85%.The main HAI site was intracranial site(n=125,64.77%),the next was lower respiratory tract (n=55,28.49%).Multivariate logistic regression analysis showed that operation grade and subtentorial operation were independent risk factors for postoperative HAI in neurosurgical patients undergoing removal of intracranial tumor,OR and 95%CI were 4.352(1.878-10.080)and 1.812(1.280-2.564)respec-tively.Conclusion Risk of postoperative HAI in neurosurgical patients undergoing high grade operation and subtentorial removal of intracranial tumor is high,effective prevention and control measures should be taken to prevent the occurrence of HAI.

Objective To explore whether hypermethylation in the promoter of p16 gene and protein of p16 were associated with development and clinicopathological characteristics of colorectal cancer. Methods Methylation-specific PCR ( MSP) and immunohistochemistry SP were used to detected hypermethylation of p16 gene and p16 protein in tumor tissues from 32 patients with colorectal cancer. Results The hypermethylation of p16 gene was detected in 40. 6% of tumor tissues. The protein of p16 promoter was detected in 75% of tumor tissues. The hypermethylation of p16 gene was detected in 63% in Dukes stages of C and D tumors. The protein of p16 promoter was detected in 69% of tumor tissues. The hypermethylation of p16 gene was detected in 25% in the stages of A and B tumors. The protein of p16 promoter was detected in 81% of tumor tissues. The hpermethylation of p16 gene was detected in 100% in low differentiated carcinomas. The protein of p16 promoter was detected in 20% , the hypermethylation of p16 gene was detected in 30% , in the high and mediate differentiated carcinomas, the protein of p16 promoter was detected in 85%. Furthermore, the hypermethylation of p16 gene was detected in 63% in the lymph node metastasis and 25% in without lymph node metastasis. The protein of p16 promoter was detected in 65% in rectum and 100% in colon. Conclusions Our study demonstrated that p16 hypermethylation and protein were associated with the development of colorectal cancer and could be used as a putative prognostic indicator for this malignancy.

OBJECTIVE To realize the situation of nosocomial infection and usage of antibiotic in order to prevent and control nosocomial infection effectively. METHODS According to the request of countrywide nosocomial infection net,using the method of clinic investigation and case history investigation,we have investigated the infection complexion about all patients in 19 May 2006 and 24 May 2007,including the duration,transferring department and dead case.Then to analyze the comparison between two results. RESULTS 3489 cases were investigated,including 186 infection cases.The prevalence rate of nosocomial infection was 5.33%.Five departments had the high prevalence rate of nosocomial infection.They were hematology department,ICU,pediatrics department,neonatal unit and cadre ward.The highest rate was 31.30%.The lowest was 0.The infection site focused on lower respiratory tract.The antibiotic utilizing rate was high. CONCLUSIONS Enhanceing the management of nosocomial infection in key departments,regulating the antibiotic utilizing,to reduce the prevalence rate of nosocomial infection.

OBJECTIVE The research aimed to investigate the incidence,clinical characteristics and risk factors of nosocomial infection in patients with mechanical ventilation in ICU,and explore the corresponding prevention procedures.METHODS Active surveillance was carried out by professional fulltime staff.The patients with mechanical ventilation in ICU above 48 hours were selected for the hospital infection surveillance,and the patients after tracheal intubation within 48 hours were also included.RESULTS The incidence of nosocomial infection was 54.28%,and the nosocomial infection were found in 69(65.71%) of the 105 cases.The major complication was mechanical ventilator-associated pneumonia(VAP).The risk factors were more than four times invasive operation and,tracheotomy(P

OBJECTIVE To explore the characteristic of nosocomial infection and formulate the effective measures of nosocomial infection management. METHODS According to the underlying disease condition and method ICD10,the infection data were to classifed and colleced which including of 160 795 cases during 2003-2006.Then the prospective and retrospective investigation were done for studying the nosocomial infection condition. RESULTS The nosocomial infection rate was 4.69%. The highest infection rate was caused by hematological disease (15.43%). By site of infection the upper respiratory infection rate was 35.34%,the lower respiratory infection rate was 28.22%,the gastrointestinal infection rate was 6.82%,and the intra-abdominal infection was 3.75%. In these infection cases,G-bacteria infection occupied 58.35% (which ranked No.1 in all pathogens),and the fungal infection occupied 17.09%. CONCLUSIONS In order to reduce the infection rate,we must enhance the work of preventing the key diseases,standard the measures of disinfection and isolation,increaseing the quarantine inspection rate and applying antibiotic according to the results of antifugal susceptibility testing.

Objective To study the influence of cochlear implantation on residual hearing in children .Methods Behavioral audiometry were performed pre -implant and 3~21 months post -implant on thirty -four cochlear implant recipients with severe to profound hearing loss .According to follow -up time ,they were divided into 2 groups which were Group A(3~12 months ,21 cases) and Group B(≥13 months ,13 cases) .The thresholds at 250 Hz ,500 Hz ,1 000 Hz and 2 000 Hz were analyzed .Results There were 25 out of 34 patients (73 .53% ) had partial residual hearing after cochlear implantation .Comparing to the hearing loss pre -operation and post -operation , which were most obvious at 500 Hz ,followed by 250 Hz ,1 000 Hz ,2 000 Hz (P<0 .05) ,and there were significant different among different frequencies .There was significant difference at different frequencies at hearing loss thresh-olds only in Group A .But there was no significant difference in Group B .With the prolonged time after the cochlear implantation ,residual hearing at all frequencies showed a trend of recovery .Conclusion The residual hearing could be partial preserved after cochlear implant in pediatric patients with severe to profound hearing impaired ,the residu-al hearing at lower frequencies (250 Hz ,500 Hz) were less affected than those at higher frequencies .With the pro-longed time after the cochlear implantation ,the residual hearing showed a certain degree of recovery .

OBJECTIVETo study the efficacy and safety of Naoxintong capsule treatment of stroke recovery with Qi-deficiency and blood-stasis syndrome (cerebral infarction), and to compared the non-inferiority analysis with the positive drug Tongxinluo capsule.

METHODTaking Tongxinluo capsules as control, randomized, double-blind, controlled, multi-center clinical experiments were studied. The evaluating indexes included the decrease of integral value of stroke patients, changes in traditional Chinese medicine, the improvement of the patient viability status (disability level), Chinese stroke scale (CSS), activities of daily living (DAL) scale and barthel index (BI ) points.

RESULTThe total effect of the two groups, Chinese and other symptoms, showed no significant statistical significance.

CONCLUSIONNaoxintong capsule stroke recovery, with Qi-deficiency and blood-stasis syndrome (cerebral infarction) has a therapeutic effect, and more secure.

Activities of Daily Living ; Adult ; Aged ; Capsules ; Double-Blind Method ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Qi ; Stroke ; drug therapy ; Treatment Outcome

Objective To study the mechanism of Wuzi Yanzong Wan (Five-Seed Progeny Pill) on improving heat stress resistance of sertoli cells in rats based on data from the whole genome microarray.Methods Sertoli cells in primary culture were randomly divided into Wuzi Yanzong Wan (WZYZW) group, normal group and model group.The cells in three groups were given WZYZW solution and equal volume of PBS respectively.After 24 hours, rats in WZYZW group and model group were placed in water bath at 43 ℃ for 15 minutes, while rats in normal group were put in water bath at 37 ℃ for 15 minutes, then total RNA of three groups were extracted respectively and data was analyzed with gene chip technology.Results Compared with the model group, there were totally 1,203 differently genes in WZYZW group based on Sertoli cell gene expression profiles, in which 697 were down regulated and 506 were up-regulated.There were totally 473 differently expressed genes in the normal group, including 168 up-regulated genes and 305 down regulated genes.Among these down-regulated gene expressions, RGD1305298, Trpt1, Dll3, Abi3 and Ppp1r12b were significant in the WZYZW group and normal group, while Dusp7 was significant among up-regulated gene expressions.The genes mentioned above were related to cell cycle, apoptosis, cell metabolism, cell adhesion and Notch signal pathway, regulation of actin cytoskeleton, MAPK signaling pathway and tight junction pathway, etc.Conclusion The mechanism of Wuzi Yanzong pill in improving heat stress resistance of sertoli cells may involve maintenance of tight junctions between sertoli cells, inhibition of sertoli cells apoptosis and promotion of cell differentiation and maturation.

ObjectiveTo discuss the effect and mechanism of Wuzi Yanzong Wan (Five Seeds for Multiplying Pills) in the proliferation of Sertoli cells in infant rats.Methods The Sertoli cells were separated from male SD rats (20 d old) and cultured,and then randomly divided into normal group,lowdose Wuzi Yanzong Wan group (5 g/L,low-dose group),mid-dose Wuzi Yanzong Wan group (10 g/L,mid-dose group) and high-dose Wuzi Yanzong Wan group (15 g/L,high-dose group).The changes of optical density (OD) of Sertoli cells were detected by using methyl thiazolyl tetrazolium test (MTT') in all groups.The expression of Ki67 positive granules in Sertoli cells were detected by using immunohistochemistry technique in all groups.The expressions of Akt and p-Akt in Sertoli cells were detected by using Western blot method.Results Compared with normal group,the proliferation of Sertoli cells treated with Wuzi Yanzong Wan was promoted significantly (P ＜ 0.05 or P ＜ 0.01).The expression of Ki67 positive granules increased significantly (P ＜ 0.05 or P ＜ 0.01),and relative expression of p-Akt increased significantly (P ＜ 0.05 or P ＜ 0.01).Conclusion Wuzi Yanzong Wan can possibly promote the proliferation of Sertoli cells in infant rats through activating Akt signal transduction pathway and up-regulating p-Akt expression.