This study was aimed to explore an effect evaluation method of traditional Chinese medicine (TCM) in the treatment of viral pneumonia. Association rules were used to analyze the therapeutic effect of heat, stagnation, and phlegm related syndromes from 297 respiratory syncytial virus (RSV) pneumonia children before and after treatment. The results showed that through the comparison of the frequent itemset of heat, stagnation, and phlegm related syn-dromes, in part of the time, the support degree of the experimental group was obviously lower than that in the con-trol group (P < 0.05). The frequent itemset of the experimental group disappeared 1~3 d earlier than the control group, which indicated the effect in the experimental group was better than the control group. It was concluded that association rules can be used for evaluating therapeutic effect of TCM in treatment of viral pneumonia. The effect evaluation method based on association rules of symptoms is more in line with the syndrome differentiation of TCM.

Objective: To study the factors effecting the expression of the reporter green fluorescent protein (GFP) gene in the mouse embryo stem cell line R1. Methods: Three different kinds of GFP euko-expression vectors were constructed, and the expression efficiency was contrasted both at mRNA and protein levels after they were integrated into the chromosomes of host cells. Results: At protein level, the GFP expression level of the colonies transfected by the expression vector-pEF-GFP with the promoter of the peptide elongation factor (EF) were significantly higher than that of the colonies transfected by pCMV-GFP with CMV promoter and by pdCMV-GFP with double copies of CMV-GFP expression unit. There was no significant difference between the colonies transfected by pCMV-GFP and pdCMV-GFP. The detection results on mRNA level of GFP had the same tendency as that at protein level. Conclusion: (1) GFP gene expression efficiency controlled by EF promoter is distinctly higher than that by CMV promoter in NIH3T3 and R1 ES cell line.(2) A slight increase of the copy number of the foreign gene expression units in the host chromosome can not make obvious increase of its expression efficiency. (3) The vector express GFP in R1 ES cell line efficiently and stablely is obtained.

Objective: To study the factors effecting the expression of the reporter green fluorescent protein (GFP) gene in the mouse embryo stem cell line R1. Methods: Three different kinds of GFP euko-expression vectors were constructed, and the expression efficiency was contrasted both at mRNA and protein levels after they were integrated into the chromosomes of host cells. Results: At protein level, the GFP expression level of the colonies transfected by the expression vector-pEF-GFP with the promoter of the peptide elongation factor (EF) were significantly higher than that of the colonies transfected by pCMV-GFP with CMV promoter and by pdCMV-GFP with double copies of CMV-GFP expression unit. There was no significant difference between the colonies transfected by pCMV-GFP and pdCMV-GFP. The detection results on mRNA level of GFP had the same tendency as that at protein level. Conclusion: (1) GFP gene expression efficiency controlled by EF promoter is distinctly higher than that by CMV promoter in NIH3T3 and R1 ES cell line.(2) A slight increase of the copy number of the foreign gene expression units in the host chromosome can not make obvious increase of its expression efficiency. (3) The vector express GFP in R1 ES cell line efficiently and stablely is obtained.

Objective To observe the influence of suspension concentrate of niclosamide on the enzyme activity of Oncomelania hupensis in order to explore its molluscicidal mechanism. Methods Oncomelania hupensis snails were collected in the habitates of river marshland in Dantu District, Zhenjiang City, Jiangsu Provence and were divided into 2 groups. The snails of the treated group were sprinkled with 25% suspension concentrate of niclosamide. The snails of the control group were sprinkled with distilled water. The soft body tissue of the snail was separated and the sections of snail tissue were made in the Cryostat Microtome. The stain of enzyme-histochemistry showed CCO, LDH, SDH, CHE and NOS had been done, and then the staining block was made by routine method. The staining reaction in the snail tissue and the average gray density were observed with the image analysis system of biomicroscope. Results The enzyme activity of CCO, LDH, SDH, CHE and NOS located in the mouth, muscle fiber, tegumentary membrane, ganglia, liver and pharyngeal cavity of Oncomelania hupensis snails. The enzyme activities of CCO, LDH, SDH, CHE and NOS in the treated group were significantly lower than that in the control group. Conclusions Niclosamide can affect the transmitting of neurohypophysis and obstruct the energy and result in the disorder of the physiological functions in Oncomelania hupensis. It is one of the reasons of Oncomelania hupensis death.

Objective To enhance the protective immunity effects against Schistosoma japonicum infection by priming with cocktail DNA vaccines and boosting with cocktail protein vaccines in infected BALB/c mice.Methods Plasmids and proteins for immunization were prepared and diluted in no bacterial saline solution to final concentration of 1.5 mg/ml,and mixed with pcDNA3.1-SjC23,pcDNA3.1-SjCTPI,pcDNA3.1-(CDR3)6 plasmid DNAs by equal volume to form the cocktail DNA vaccine,and also mixed with recombinant proteins SjC23-HD,SjCTPI,and NP30 by equal volume to form the cocktail protein vaccine.Seventy female BALB/c mice of 4-5 weeks old were randomly divided into 5 groups(A,B,C,D,E).In Group A(control group),each mouse was immunized with 100 ?l saline solution by intramuscular(i.m.);in Group B(pcDNA3.1 control group),each mouse was immunized(i.m.)with 100 ?l pcDNA3.1 for three times at week 0,3,6;in Group C(pcDNA3.1 and cocktail protein group),each mouse was immunized(i.m.)with 100 ?l pcDNA3.1 for three times at week 0,3,6 and immunized with 100 ?l mixed protein vaccines plus 100 ?l FCA by subcutaneous at week 9;in Group D(cocktail DNA vaccines group),each mouse was immunized(i.m.)with 100 ?l mixed DNA vaccines for three times at week 0,3,6;in Group E(cocktail DNA vaccines plus cocktail proteins),each mouse was immunized(i.m.)with 100 ?l mixed DNA vaccines for three times at week 0,3,6 and immunized with 100 ?l mixed protein vaccines plus 100 ?l FCA by subcutaneous at week 9.Four weeks after the last DNA immunization or two weeks after protein boosting,all the mice were challenged with(40?1)cercariae of Schistosoma japonicum by abdominal skin penetration at the same time.Forty-two days post-challenge,the mice were sacrificed and perfused,and the numbers of recovered worms and eggs in liver were counted.The blood was collected from the tail veins of all the mice two days before the first immunization and challenge,respectively,the serum was prepared for detection of IgG,IgG1 and IgG2a.Two days before the challenge,the spleen cells of two mice from each group were cultured and stimulated with ConA and soluble egg antigen(SEA),and the supernatant was collected for detection of IL-2,IL-4 and IFN-?.Results The worm reduction rates in Group C,D and E were 17.70%,32.88% and 45.35%,respectively,compared with the control group.The worm reduction rates in Group D and E were significantly higher than that in Group C(P

Objective To explore the Schistosoma japonicum cercariae collected method and the quantity of cercariae obtained from a great quantity artificial infected snails. Methods In laboratory condition, Oncomelania snails were infected with schistosome miracidia. Sixty days post-infection all snails were divided into 7 shares. Cercariae shed from 1 share snails every day and the number of all shedding days were 40. Cercariae shed from snails were collected with low-velocity centrifuge and the cercariae sediment were weighted. Results One thousand and nine hundred g snails bred for 120 days post-infection, the infection and survival rates were 36. 00% and 51. 58%. Cercariae col-lected were 10. 5 g from 40 days collection. Cercariae quantity of shedding from 1 000 positive snails per day was 0. 257 4 g.

Objective To observe the impact of water temperature on shedding and infectivity of the cercariea of Schistosoma japonicum under laboratory condition. Methods Infected snails were exposed to 1, 5, 10, 15, 20, 25, 30, 35℃ and 40℃ water for 4 hours for shedding of cercariae respectively, the shedding rates and total numbers of cercariae shed were investigated. The mice were infected with cercariae in 1, 10, 20, 30℃ and 40℃ water for 30 minutes respectively, the infection rates and mean worm burden of mice were investigated. Results The cercariae were shed from infected snails with Jiangsu or Yunnan isolate of [WT5”BX]S. japonicum in water ranged from 1℃ to 40℃, but the fittest water temperatures range for shedding of cercariae of Yunnan isolate of S.japonicum[WT5”BZ] is from 20℃ to 35℃, while fittest range for Jiangsu isolate is from 15℃ to 35℃. All of the infection rates of mice infected with cercariae of Jiangsu isolate of S.japonicum in the water ranged from 1℃ to 40℃ were 100.0%. Infection rates of mice infected with cercariae of Yunnan isolate of S. japonicum in the water ranged from 1℃ to 40℃ were more than 83.3%. Conclusions When infected snails are exposed to the water with temperature ranged from 1℃ to 40℃ it is possible for cercariae to shed and infect the final hosts . It is suggested that there is a possibility of infection with S.japonicum when contacting the water near marshlands with infected snails in winter.

Objective To investigate the clinical effect of Ethibond suture and anchor fixation by arthroscopic technique for the treatment of avulsion fracture of the anterior cruciate ligament (ACL) tibial insertion.Methods Twenty patients with avulsion fracture of the ACL tibial insertion hospitalized between July 2013 and June 2014 were collected retrospectively.There were 12 males and 8 females,aged 18-41 years (mean,25.3 years).All patients were identified with type Ⅱ and type Ⅲ fractures according to the Meyers-McKeever classification.Results of Lachman test and anterior drawer test were both positive.All patients accepted the operation that avulsion fracture of the ACL tibial insertion was treated with the Ethibond suture and anchor fixation by arthroscopic technique within 3 weeks after injury.Follow-up X-ray examinations were carried out to evaluate the bone union.Lysholm and international knee documentation committee (IKDC) scores were used to evaluate the function of knee joint postoperatively.Results Operation time was 45-70 min (mean,50 min).Blood loss was 5-15 ml (mean,10 ml).Follow-up was conducted for 12-24 months (mean,16.3 months).Postoperative X-ray showed the reduction was satisfactory.Lachman test and anterior drawer test were both negative after operation.Knee functions were recovered to normal.Lysholm and IKDC scores were 89-96 points [(93.5 ± 2.3) points]and 84-96 points [(91.0 ± 3.9)points] respectively at the final follow-up,improved compared to the preoperative 42-61 points [(51.1 ± 6.2) points] and 46-68 points [(55.2 ± 7.0) points] (both P ＜0.05).Conclusion Arthroscopic Ethibond suture and anchor fixation for avulsion fracture of the ACL tibial insertion is an effective procedure with the advantages of small trauma,reliable fixation,good functional recovery and satisfactory clinical effect.

Objective To understand whether the breeding time of snails influences the evaluation on molluscicidal efficacy against Oncomelania snails so as to make a standardization of methods for screening molluscicides in laboratory. Methods Snails collected from the endemic areas in Nanjing and Tongling cities bred for 1,6,11,16,21.31,61 and 91 days respectively, and then were immersed in 1.000 0,0.500 0,0.250 0,0.125 0,0.062 5 and 0.031 3 mg/L solution of niclosamide for 24 and 48 hours at 25℃ in laboratory. Results One hundred percent killing rate was achieved in the groups of the snails bred for 1 - 11 days and immersed in 1.0 mg/L niclosamide for 24 hours. The LC_(50) concentrations of snails collected from Nanjing, Jiangsu Province and immersed for 24 hours varied from 0.094 7 to 0.133 9 mg/L, and for 48 hours from 0.071 8 to 0.092 6 mg/L.Those of snails collected from Tongling, Anhui Province for 24 hours varied from 0.082 5 to 0.103 9 mg/L, and for 48 hours from 0. 071 8 to 0.082 5 mg/L. The molluscicidal effect was unstable in the groups of snails bred for more than 16 days. There was a significant difference (X_(Nanjing24 h)~2 = 22.26,P

OBJECTIVETo examine the differential levels of fecal Bifidobacterium, Bacteroides, Eubacterium rectale-Clostridium, Escherichia coli, Enterococcus, and Clostridium difficile between patients with hepatic cirrhosis and healthy controls. Fecal samples were collected from 29 patients with hepatic cirrhosis treated in the Department of Digestive Diseases at Zunyi Hospital between March and December of 2010.

METHODSFecal samples were collected from 13 healthy college students for use as controls. All samples were assessed by pH measurement, bacterial culture for turbidity, fluorescence in situ hybridization, and laser scanning confocal microscopy. The t-test and rank correlation test were used to determine statistical significance of intergroup differences in each tested parameter.

RESULTSThe feces of patients with hepatic cirrhosis had higher pH than that of healthy controls (6.79+/-0.64 vs. 6.18+/-0.74, P less than 0.05). The bacterial turbidity was not significantly different between the feces of hepatic cirrhosis patients and healthy controls (1.15+/-0.59 vs. 1.39+/-1.01, P more than 0.05). The numbers of Bifidobacterium, Bacteroides, Eubacterium rectale-Clostridium, Escherichia coli, Enterococcus, and Clostridium difficile in feces of patients with hepatic cirrhosis were significantly lower than those of the controls (all P less than 0.01). No significant correlation was found between the number or ratio of bacteria species and the severity of hepatic cirrhosis (Child-Pugh scores; P more than 0.05).

CONCLUSIONThe total quantity of intestinal bacteria in patients with hepatic cirrhosis is not significantly different from that in healthy patients. However, the profile of intestinal bacteria is different, which may explain the increased pH of fecal samples from patients with hepatic cirrhosis, but the differential profile is not correlated to cirrhosis pathogenesis.

Adult ; Aged ; Aged, 80 and over ; Bacteroides ; isolation & purification ; Bifidobacterium ; isolation & purification ; Case-Control Studies ; Clostridium ; isolation & purification ; Enterobacteriaceae ; isolation & purification ; Feces ; microbiology ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Liver Cirrhosis ; microbiology ; Male ; Middle Aged ; Young Adult