Cryptococcosis ; Groin ; Humans ; Lymph Nodes ; microbiology ; Lymphadenitis ; microbiology

Objective To develop a TaqMan MGB probe-based,sensitive and specific real-time fluorescence quantitative PCR assay for rapid detection of Helicobacter hepaticus.Methods Primers and probes specific toflaB gene of Helicobacter hepaticus were designed.A TaqMan MGB probe-based,real-time fluorescence quantitative PCR was established.The specificity,sensitivity and stability of the assay were assassed.Then,the established TaqMan MGB probe real-time fluorescence quantitative PCR assay was applied to detect Helicobacter hepaticus in 1081 clinical specimens during 2008-2011,compared with bacterial isolation and culture method and conventional PCR assay.Results The specificity of this established TaqMan MGB probe-based real-time fluorescence quantitative PCR was high and there were no cross-reactivity with Helicobacter pylori,Campylobacter jejuni,Clostridium piliforme,Pasteurella pneumotropica,Escherichia coli,Pseudomonas aeruginosa.The detection limits was 8.3 copies.The correlation coefficient and slope value of standard curve were 0.999 and -3.227,respectively and the efficiency of TaqMan MGB-based probe realtime fluorescence quantitative PCR assay was 100％.The TaqMan MGB-based probe real-time fluorescence quantitative PCR and conventional PCR were preformed to detect Helicobacter hepaticus in 1081 clinical specimens,a total of 86 specimens were positive for Helicobacter hepaticus.However,there was only 4 specimens were positive by bacteria isolation and culture method.The results showed that TaqMan MGB -based probe real-time fluorescence quantitative PCR for Helicobacter hepaticas was more sensitive than bacteria isolation and culture method,and it could detect Helicobacter hepaticus DNA from clinical specimens directly,and detection time is only 2 hours.Conclusion The TaqMan MGB-based probe real-time fluorescence quantitative PCR assay was a reliable,specific,sensitive and useful tool for rapid detection of Helicobacter hepaticus.

Objective To explore the effect of Jiangzhipailuan decoction in regulating PPARα( belong to the nuclear receptor family of ligand-activated transcription factors ), PGC- 1α ( peroxime proliferator activated releptour)and SREBP-1c( belong to the baichelix-loop-helix-leucine zipper class of transcription factors), SCAP( SREBP cleavage activating protein) related to lipid metabolism in the treatment of polycystic ovarian syndrome. Methods 84 patients suffered polycystic ovary syndrome were randomly divided into 4 groups: one received traditional Chinese medicine treatment, one western medicine treatment, one combination therapy and one as the control group. Traditional Chinese medicine group was treated with Jiangzhipailuan decoction treatment for 3 months, western medicine group was treated with up to Diane-35 ( ethinyl cyproterone tablets) for three cycles, while the combination therapy group was treated with traditional Chinese medicine ,western medicine as well as combined treatment for 3 courses. Results In the combination therapy group the PPARα, PGC-1α and decreased SREBP-1c, SCAP copy number was significantly improved ,and body mass index was significantly lowered. The total improvement rate of menstrual in three groups were 71.42% ,75.00% ,92.86% respectively. Conclusion Jiangzhipailuan decoction played a prominent role in regulating PPARα,PGC-1α and SREBP-1c,SCAP related to lipid metabolism in the treatment of polycystic ovarian syndrome.

Purpose To analyze the clinical pathological characteristics and pathological diagnosis and prognosis of spindle cell carci-noma of the breast. Methods Three cases of spindle cell carcinoma of the breast were studied by morphological and immunohisto-chemical EnVision techniques. Results The females were 48, 63 and 71 years old. The tumors located in the right breast with 4. 0 cm × 3. 0 cm × 3. 0 cm, 3. 0 cm × 2. 0 cm × 2. 0 cm and 3. 5 cm × 2. 8 cm × 2. 3 cm in size and showed cystic lesion. The neoplasm was composed of bland spindle cells and mimicking fibromatosis. Immunohistochemical staining showed that spindle cells were positive for CK(AE1/AE3), CK(34βE12), CK14, CK5/6, p63 and vimentin, negative for ER, PR and c-erbB-2. Ki-67 was positive in 20%, 25% and 20% of the cells. Conclusion Spindle cell carcinoma of the breast is a rare subtype of the metaplastic carcinoma which tend to show cystic changes. It is important to make a definite diagnosis which combine histopathologic features and immunophe-notyping.

Objective To improve the accuracy of detection through analyzing the phenotypes of P.pneumotropica isolates in laboratory animals in Beijing area .Methods 306 suspicious P.pneumotropica strains were identified by biochemical identification and 16S rDNA sequencing.Then, to obtain the phylogenetic relationships combined with colony characteristics on blood agar plates and biochemical characteristics of 53 biotypes .Results BD Phoenix 100 automated bacterial identification system and 16S rDNA sequencing identified P.pneumotropica positive rate of 306 isolates were 164/306 and 227/306, respectively.There were 140 phenotypes in 227 true-positive strains, of which 106 were biotype Heyl and 23 were biotype Jawetz .Conclusions In the samples of laboratory animals in Beijing area , P.pneumotropica infection mainly are of biotype Heyl , and less is of biotype Jawetz .The phenotypes are diverse and widely distributed .

Objective To develop a multiplex polymerase chain reaction ( mPCR) assay for detection of four pathogenic dermatophytes [Trichophyton mentagrophytes (Tm), Microsporum gypseum (Mg), Microsporum canis (Mc), and Arthroderma simii ( As) ] in laboratory animals, which could be used rapidly and simultaneously for direct detection of those four pathogens.Methods We designed 5 specific primers according to 18S-28S rRNA sequences of the four pathogenic dermatophytes reported in Genbank. The four mPCR assays were established through optimizing the concentration of primers, dNTP, TaqDNA polymerase and the annealing temperature.After verifying the specificity and sensibility, this method was used to detect 15 hair samples with artificial infection and 260 samples taken from laboratory animals.Results This mPCR technique can distinguish the four dermatophytes by producing 192 bp( Tm) ,460 bp( Mg) , 290 bp( Mc) and 602 bp( As) fragments.The sensibility for detection of the four dermatophytes was 5.9 pg/μL, 6.6 pg/μL, 9.5 pg/μL and 5.1 pg/μL, respectively.The results of 15 artificial infection samples were accurate, and the results of 260 hairs samples were negative for the four fungi.Conclusions Our results suggest that the mPCR assay developed in this study can efficiently detect the four dermatophytes, is a useful and rapid technique for rapid detection of the pathogenic dermatophytes in laboratory animals.

With this retrospective study, we analyzed the negotiation subject, drug type, drug access mechanism, negotiation link and negotiation result of drug negotiation in typical regions at home and abroad.In foreign countries, drug negotiation mechanism has been implemented for many years, and many countries have established a relatively perfect negotiation system.We selected 8 typical countries and regions for which we statistically analyzed the negotiation subject, drug type, drug access mechanism, negotiation link and negotiation result of drug negotiation and these include the United States, Australia, Canada, France, Germany, Italy, South Korea and Taiwan.However, in recent years some Chinese medical insurance department carried out pilot works on drug negotiations, and they got some successful results of the implementation of practical experience.Eight typical Chinese regions were also selected for statistical analysis, and these include Zhejiang, Hunan, Jiangsu, Jiangxi, Chengdu and Qingdao.From the analysis of the comparison of drug negotiation mechanism in domestic and foreign typical regional, we found that foreign regional drug negotiation mechanism is more mature and perfect, while in the domestic areas the mechanism is still poor at a certain extent as compared to foreign countries.We should learn from the successful experience of foreign countries and also establish and improve the negotiation mechanism that is suitable for China''s national conditions.

Objective To establish an effective PCR assay for leptospirosis detection , and applicate the assay in tree shrew, mongolian gerbil and gray hamster .Methods Sequence of leptospira was obtained from the NCBI Genbank , and primers were designed based on the sequences .The positive amplified fragments were sequenced to verify the reliability of the method.The samples from tree shrew, mongolian gerbils and hamsters were tested using this PCR method .Results The PCR method for detection of leptospirosis was successfully established .The positive rate of Leptospira was 8.33% in 60 samples of conventional tree shrews , 100% in 104 samples of the conventional Mongolian gerbils , and 0% in 60 samples of clean gray hamsters.Conclusions The establishment of this PCR assay is useful in the detection of leptospirosis in tree shrew, mongolian gerbil and gray hamster .The results of our investigation of leptospira infection levels of the three new experimental animals may promote their application in biomedical research .

Objective To evaluate the effects of delayed cord clamping (DCC) on preterm infants with gestational age ＜32 weeks.Methods Literatures from January 1,1990 to April 30,2013 in Cochrane library,PubMed,EMBASE,China Academic Journal Network Publishing Database,Wanfang Medical Database and VIP Database were searched.Randomized controlled trials (RCT) of DCC in preterm infants with gestational age ＜32 weeks were screened and evaluated.DCC was defined as cord clamping in 30-90 s after delivery,and early cord clamping (ECC) (＜30 s) was as the control.Rev Man 5.1.0 was used for statistical analysis.Mean difference (MD) and 95％CI were used for continuous data while OR and 95％CI were for categorical data.Results Nine studies (11 articles) involving 373 infants were included.Compared with ECC,DCC improved hematocrit (MD=4.19,95％CI:2.97-5.40,Z=6.74,P＜0.000 01),blood volume (MD=11.70,95％CI:6.02-17.38,Z=4.04,P＜0.0001) and mean arterial pressure of preterm infants with gestational age ＜32 weeks (MD=3.11,95 ％CI:1.30-4.92,Z=3.37,P=0.0008),decreased the usage of volume expansion for hypotension (OR=0.32,95％CI:0.11-0.98,Z=2.05,P=0.04) and the incidence of necrotizing enterocolitis (OR=0.48,95％CI:0.25-0.92,Z=2.22,P=0.03).Meanwhile,DCC had no influence on the peak bilirubin concentration,the incidence of sepsis,patent ductus arteriosus,retinopathy and intracranial hemorrhage,also no influence on neonatal mortality on dcscharge,mental developmental index and psychomotor developmental index at seven-month old.Conclusions DCC might be a safe procedure to improve prognosis of preterm infants less than 32 weeks' gestational age.However,due to small sample size and lack of data on follow up,it is necessary to launch clinical trials with higher quality and larger scale to further evaluate the effect and safety of DCC.

Objective To establish a detection technique for H.pylori(HP) infection in Mongolian gerbils using nested PCR technique.Methods H.pylori was cultured in vitro and inoculated into Mongolian gerbils.At the 10th week after infection, the HP in the gastric juice of Mongolian gerbil was detected by conventional PCR assay and the gastric juice, gastric mucosa, duodenal contents and colon stool were examined by nested PCR.Rapid urease test and ELISA were used to analyze the accuracy of the nested PCR assay.All of the PCR products were verified by sequencing.Results The positive rate of gastric juice detected by conventional PCR was 30%, while the positive rates of gastric juice, gastric mucosa, duodenal contents and colon stool detected by nested PCR were 100%, 100%, 90%, and 10%, respectively.The positive detection rates of rapid urease test and serum ELISA were 100% and 0%, respectively.Comparing the results of different methods, both the positive rates of gastric juice and gastric mucosa detected by nested PCR and the detection rate of rapid urease test were 100%, but the results of conventional PCR detection of gastric juice, the nested PCR detection result of stool in colon and of serum ELISA assay were lower than other methods.Conclusions Due to its high accuracy and sensitivity, the nested PCR assay of gastric juice can be used for the long-time detection of H.pylori infection in Mongolian gerbils, especially useful in the experiments of prevention and treatment of H.pylori infection.