OBJECTIVE: To establish a bacterial endotoxin test method for fluorescein sodium injection.METHODS: Based on the bacterial endotoxin test method stated in Chinese Pharmacopoeia 2005 edition(second part),an interference test was conduced on different batches of fluorescein sodium samples using tachypleus amebocyte lysate from different companies.RESULTS: At a concentration of no more than 0.75 mg?mL-1,fluorescein sodium showed no interfere with the bacterial endotoxin test.CONCLUSION: It is feasible for bacterial endotoxin test of fluorescein sodium injection be conducted in a limit value of 0.15 EU?mL-1.

Objective To study the feasibility of applying extrusion-puffing technique to the preparation of Qingyin Teabag.Methods Extrusion-puffing techique and water decoction were used to prepare the Qingyin Teabag,and thecorresponding products were compared by pharmacodynamic tests .Meanwhile,a dissolution test was carried out for the monarch drug Radix Glycyrrhizae in the teabag before and after pressurizing-expansion,thus to explore the influence of extrusion-puffing technique on the dissolution speed of ammonium glycyrrhetate from Radix Glycyrrhizae.Results With ammonium glycyrrhetate as the index,T50 and Td of Radix Glycyrrhizae before extrusion puffing extraction were 20.25 min and 56.20 min,while were 7.975 min and 18.975 min respectively after extrusion puffing extraction,the difference being very significant.Pharmacodynamic tests showed effects of the products processed by extrusion-puffing technique were slightly superior to those by water decoction.Conclusion It is feasible to adopt extrusion-puffing technique to the preparation of Qingyin Teabag.

Objective To establish a method for the determination of berberine hydrochloride in Huanglian Jiangtang Pill. Methods The sample was extracted with hydrochloric acid-methanol(1 ∶ 100). The chromatographic conditions were as follows:Diamonsil C18 chromatographic column(250 mm? 4.6 mm,5 ? m)with a mobile phase of acetonitrile-0.05 mol/L potassium dihydrogen phosphate(28 ∶ 72),the detection wavelength at 350 nm and the flow rate being 1.0 mL? min-1. Results A good linearity was obtained from 0.244 ? g to 2.928 ? g of berberine hydrochloride in Huanglian Jiangtang Pill(r=0.999 9,n=7).The average recovery was 99.07 % and RSD=1.30 %(n=6). Conclusion This method is simple,sensitive,specific and accurate for determination of berberine hydrochloride in Huanglian Jiangtang Pill.

ObjectiveTo establish nude mouse tumor models bearing human pancreatic adenocarcinoma SW1990 cells in multiple sites at different time-points and investigate the feasibiilty of multiple tumor-bearing in these models; then the findings and detection rate of 3.0T magnetic resonance image (MRI) in subcutaneous transplanted tumors was analyzed. Methods A total of 6 BALB/C nude mice were randomized into 3 groups (2 mice per group ).At the 1st,8th,15th day,the mice were injected subcutaneously with the suspension of SW1990 cells at left axilla and right axilla and right groin in sequence.Three weeks later,all the bearing-tumor mice were performed with MRI non-contrast enhanced scanning plus Gd-DTPA enhanced scan and the subcutaneous masses were subjected to pathological analysis.ResultsAll the 6 nude mice were alive during the study and obvious mass was observed in every injected site.The tumor size was positively associated with the grwing time.There were 9 tumors which could be de.ted by noncontrast enhanced MRI scanning and one more tumor was detected by contrast enhanced scanning.2 tumors were not detected,the 2 tumors were located at subcutaneous of right groin,with the shortest growing time,and the major axis of the 2 un-detected tumors was less than 5mm.Despite the MRI findings of the transplanted masses similar to that of human pancreatic adenocarcinoma with bleeding,necrosis,they presented the characteristics of a clear rim,with pseudocapeule sign.All the 12 masses were similar with human pancreatic adenocarcinoma under light microscope.ConclusionsIt is feasible to transplant human pancreatic adenocarcinoma cell at three different subcutaneous sites (injected at three different points of time) in the nude mouse,with a minimal survival time of three weeks.However,routine 3.0T MRI cannot detect the early tumors (growing time within 1 week,major axis ＜5 mm).

Objective To validate the association between genetic polymorphisms of nucleotide binding oligomerization domain 2 (NOD2) gene and the risk of tuberculosis in Chinese population.Methods A validation case-control study was performed with a total of 1043 pulmonary tuberculosis patients and 808 healthy controls.All controls had no history of tuberculosis or malignancy,and were matched with cases by sex and age.Genomic DNA from the peripheral blood samples of participant was extracted.Single nucleoside polymorphisms (SNP) of rs3135499,rs7194886,rs8057341,and rs9302752 in the NOD2 gene were genotyped using a TaqMan-based allelic discrimination system.The chi-square test was used to analyze categorical data and Logistic regression model was used to calculate the odds ratio (OR) and 95％ confidence interval (CI).Results When all patients as the cases were included in the analysis,no significant association was found between the four SNP of NOD2 gene and the risk of pulmonary tuberculosis.In a subgroup analysis by restricting cases to sputum culture positive patients,the variant genotypes of rs7194886 were significantly associated with an altered risk of tuberculosis.Compared with the CC genotype,individuals carrying the CT/TT genotype of rs7194886 had an increased risk (OR =1.35,95％ CI:1.05-1.72) for sputum culture positive tuberculosis.After adjusted for potential confounders,the increased risk of CT/TT genotype was still observed (OR=1.35,95％CI:1.05-1.73).Stratification analysis revealed that the effect of the SNP rs7194886 (CT/TT vs TT) was significant among men (OR=1.44,95％CI:1.08-1.92),individuals with age ＜55 years (OR=1.51,95％CI:1.05-2.16),smokers (OR=1.53,95％CI:1.07-2.18),and alcohol drinkers (OR=1.84,95％CI:1.01-3.33).But the heterogeneity test was not significant.Haplotype analysis showed that the rs9302752C-rs7194886T haplotype was associated with an increased risk of sputum culture positive tuberculosis (x2 =4.27,P=0.039).Conclusion In summary,genetic polymorphisms of rs7194886 in the NOD2 gene may be associated with the risk of pulmonary tuberculosis in the Chinese population.

ObjectiveTo observe the clinical effects of locking proximal humerus plate(LPHP) in the treatment of complex proximal humeral fracture of aged patients and to analyze the influencing factors. Methods Among 39 cases with complex humeral fractures (aged 60-83 years),there were 29 cases with three-part fractures and 10 with four-part fractures according to the Neer classification.They were operated by LPHP via deltoid-pectoral approach. Neer numerical rating system was employed to evaluate postoperative function of shoulders.Results39 cases were followed up for average of 16 months.According to Neer numerical rating system,the excellence rates of three-part fractures and four-part fractures were 86.2％ (25 cases) and 50.0％ (5 cases),respectively,with total excellence rate of 76.9％(30 cases). Age (OR =1.314, P＜0.05) and fracture type ( OR =1.295, P＜0.05)ofpatientswereindependentriskfactorsforprognosis of proximal humeral fracture of aged patients by multiple logistic regression. Conclusions LPHP is an effective implant for treating complex proximal humeral fracture of aged patients,with age and fracture types as important risk factors of prognosis.<英文关键词>=humeral fractures

Objective To optimize the preparation process of Tangzu Granula. Methods With the paste-forming rate and total content of paeoniflorin as the indexes, the optimal water-extraction condition was screened by orthogonal design. With hydroscopicity, fluidity and formability as the indexes, the appropriate ratio of the excipient and the formula were selected. Results The optimum water-extraction condition was as follows: 10-fold water, decoction for 2 times and 1.5 hour for each time. The optimal ratio of the excipient and the formula for preparation of Tangzu granules were two proportions of the extract and three proportions of the excipient which consisted of lactose and dextrin(2 : 1, w/ w). Conclusion The technology is effec-tive, scientific, reasonable, and practical.

Objective To evaluate 99mTc-ciprofloxacin (Infecton) scintigraphy as a method for detecting secondary infections associated with ANP in swine,in comparison with CT.MethodsTwenty-eight healthy swine were randomly assigned to control group (n =6),non-infected ANP (n =6) and infected ANP group( n =16).ANP model was induced by retrograde injection of sodium taurocholate and pancreatic protease mixture into the biliary and pancreatic duct.Two days after ANP induction,swine in infected ANP group were injected with 3 x 108 E.coli into pancreatic tissue,while swine in non-infected ANP group were injected with inactivated E.coli.At 7 d after inoculation,at 0.5,1,2,3,4,and 6 h after intravenous administration of 370 MBq of Infecton,SPECT scan was performed.Then 64-slice spiral CT scan was performed.Then swine were sacrificed,and histopathology examination and bacterial culture of pancreatic tissue were performed.The sensitivity,specificity,accuracy,positive predictive value and negative predictive value of the two methods to detect secondary infections were determined.ResultsThere were no abnormality in the normal pancreas and the bacterial culture was negative.There were pancreatic necrosis in the non-infected ANP group,but the bacterial culture was negative.There were pancreatic necrosis and infection in the infected ANP group and the bacterial culture was positive.The sensitivity,specificity,accuracy,positive predictive value and negative predictive value of the Infecton method were 93.8％ ( 15/16 ),91.7％ ( 11/12 ),92.9％ ( 26/28 ),93.8 ％(15/16) and 91.7％ ( 11/12),whereas these values for CT were 12.5％ (2/16),100.0％ ( 12/12),50.0％(14/28),100.0％ (2/2) and 46.2％ (12/26),respectively.The sensitivity,accuracy,and negative predictive value of the Infecton method were significantly higher than those in CT group (P ＜0.01 ).ConclusionsInfecton scintigraphy may be a better procedure for detecting ANP secondary infections than CT.

Objective To study the potential and value of ultrasmall superparamagnetic iron oxide (USPIO) conjugated by mesothelin antibody as MRI targeting contrast agent for diagnosis of implanted human pancreatic carcinomas in nude mouse.Methods Nude mouse tumor models bearing multiple human pancreatic carcinomas at different time points was established and they were randomized into two groups,and USPIO or MSLN-USPIO were used as contrast enhanced agents in the 3.0T MRI scan,respectively,then the positive detection rates for smallest tumors,and the signal intensity of tumors in T2 mapping images of both unenhanced and contrast enhanced scanning and the negative enhancement rate were measured,then Prussian blue staining was performed in alI the tumor specimens to observe the difference of Fe3 + ion deposition.Results There was no statistical significance between USPIO group and MSLN-USPIO group in the positive detection rates for smallest tumors.In USPIO group,the negative enhancement rate of left or right axilla tumors was (12.29 ±7.45)％ and (11.06 ±5.91)％,and they were (33.88 ±6.09)％ and (43.29 ± 11.64)％ in MSLN-USPIO group.There was statistical significance in the difference of signal intensity between unenhanced and contrast enhanced in left or right axilla tumors (P ＜ 0.05),and the negative enhancement rate in MSLN-USPIO group was significantly higher than that in USPIO group (P ＜0.05).The Fe3+ ion deposition in tumors' tissue in MSLN-USPIO group was significantly more than that in USPIO group.Conclusions The enhanced effect of MSLN-USPIO is superior to USOPIO,and it can be a tumor targeted MR contrast enhanced agent for the diagnosis of pancreatic carcinoma in nude mouse.

Objective To assess the prevalence of Campylobacter in the children with acute bacterial diarrhea in Shanghai.Methods Epidemiological survey.Totally 6 641 children with acute bacterial infectious diarrhea from outpatients and inpatients in Children′s Hospital of Fudan University were submitted to the investigation during January 2011 to December 2012.The Campylobacter was isolated from stool samples collected from subjects in micro aerobic environment and identified by both multi-PCR and matrix-assisted laser desorption ionization-time-of-flight mass spectrometry ( MALDI-TOF MS ) .Antimicrobial susceptibility tests were assayed by disk-diffusion method according to EUCAST standard.The isolates molecular typing was done by PFGE.SPSS16.0 was used to analyze the results.Results A total number of 6 641 subjects were enrolled, among them, 305 patients were infected with Campylobacter.The prevalence rate was 4.6%(305/6 641).Among the infected patients, 240 patients were infected with Campylobacter jejuni and 65 patients were infected with Campylobacter coli, the infectious rates of these two pathogenic bacteria were 3.6%and 1.0%, respectively.The peak infectious rate in patients older than 1 year of age was 6.2%(209/3 385) which was higher than that in children under 1 year of age (2.9%, 96/3 256),χ2 =35.98,P<0.001.The infectious rate in winter and spring (6.8%, 138/2 040) was higher than that in the other seasons ( 3.6%, 167/4 601 ) ,χ2 =28.59, P <0.001.Antimicrobial susceptibility test results showed that 91.5%( 279/305 ) isolates were resistant to ciprofloxacin and 11.8%( 36/305 ) isolates were resistant to erythromycin.A total of 9 genotypes of Campylobacter were found by PFGE cluster analysis.The similarity were ranged from 65.1%-100.0%for type A, 67.6%-100.0%for type B, 61.7%-100.0%for type C, 59.0%-100.0%for type D, 71.4%for type F, 80.0%for type H, 54.4%-90.9%for type I, and only one strain was classified as type E and G.Conclusions Campylobacter is a major pathogenic bacteria associated with acute bacterial infectious diarrhea in children especially in children older than 1 year of age in Shanghai.The prevalent pattern of this pathogen was sporadic and the sharp peak was in winter and spring.The isolates are highly resistant to ciprofloxacin but still sensitive to erythromycin.