Objective To develop a multiplex polymerase chain reaction (PCR )method for detecting and genotyping moxifloxacin-resistant Clostridium difficile (C.difficile)isolates.Methods Specific PCR primers of slpA genotypes gr,hr,fr,gc08 and 078 were designed according to the differences of slpA nucleotide sequences in different C.difficile genotypes,and the house-keeping gene tpi specific PCR primers were also added for the construction of multiplex PCR method.Nine common intestinal normal and pathogenic strains were used to verify the specificity of slpA multiplex PCR for the detection of C.difficile.Forty-six C.difficile reference strains,belonging to 11 slpA genotypes,were used to verify the ability of the multiplex PCR method for dectecting and genotyping.Thirty-nine moxifloxacin-resistant clinical isolates were genotyped by the multiplex PCR,and its clinical value was evaluated by comparing with slpA sequence typing (slpA ST)method.Results All the 9 intestinal normal and pathogenic strains were negative when detected by the multiplex PCR.And tpi of 46 C. difficile reference strains were positive,and 36 strains belonging to slpA genotypes gr,hr,fr,gc08 and 078 were genotyped correctly.Other 10 strains which belonged to other 6 genotypes were non-typeable. Among 39 moxifloxacin-resistant clinical isolates,all were positive of tpi,and 32 isolates were typed correctly by the multiplex PCR method,including 22 slpA genotypes gc08,6 genotypes hr,2 genotypes fr,and 2 genotypes 078,which were consistent with slpA ST.However,7 isolates could not be typed by multiplex PCR,which were identified as other genotypes not included in the multiplex PCR by slpA ST. Conclusions A convenient and rapid multiplex PCR method for the detection of C.difficile is established successfully,which can distinguish among five slpA genotypes.slpA genotype gc08 is the common genotype of moxifloxacin-resistant clinical isolates.

Objective To evaluate the diagnosis and treatment of cystic renal cell carcinoma and to improve the preoperative diagnosis and curative rate of the disease. Methods Ten cases of cystic renal cell carcinoma were retrospectively analyzed in the aspects of imaging and pathologic characteristics. There were 7 males and 3 females with average age of 56 years old (ranging from 38--74 years old) in this study. There were 3 cases complained of sore waist, 7 cases were found renal masses in annual physical examination and 2 cases had the history of renal cysts. The cyst diameter was 3.5 8.2 cm. Six cases had been diagnosed with ultrasound and 7 cases had been diagnosed with CT scan pre-operatively. Eight eases were diagnosed with frozen section during operation. All the 10 cases accepted radical nephreetomies. Results The post-operative histological diagnosis showed that there were 9 cases of clear cell carcinoma and 1 case of granular cell carcinoma. The pathological character istics were tumor necrosis of renal cell carcinoma in 6 cases, multilocular cystic renal cell carcinoma in 2 cases and carcinoma in renal cyst in 2 cases. Eight patients followed up from 6 months to 5 years. Six patients were still alive (mean 28.5 months). Conclusion The keys to improve the diagnosis and curative rate of the cystic renal cell carcinoma are paying attention to the pre-operative imaging study, the intra-operative frozen section examination and histopathology results.

Objective To compare the effect of retroperitoneoscopic ureterolithotomy (RPUL) and ureteroscopic lithotripsy (URL) for upper ureteral calculi.Methods One hundred and twenty cases treated by RPUL and 108 cases by URL from January 2002 to October 2012 were reviewed.In RPUL and URL group,the diameter of stone was (1.56 ± 0.52) cm vs (1.44 ± 0.46) cm,ipsilateral hydronephrosis was (2.85 ± 0.86) cm vs (2.76 ± 0.82) cm,body mass index was (23.65 ± 2.80) kg/m2 vs (22.54 ± 2.68) kg/m2.There were no signficant differences.Data on the operation time,the hospital stay after operation,the operation,successsful rate,complication incidence and stone-free rate were compared between the 2 groups.Results Comparisons between RPUL group and URL group included the following:the operation time was (75.5 ± 25.8) min vs (62.5 ± 15.3) min,the hospital stay after operation was (6.2 ± 1.2) d vs (4.0 ± 0.8) d.There were significant differences.The operation successful rate was 95.0％ (114/120) in RPUL group and 85.2％ (92/108) in URL group.The complications incidence rate was 3.5％ (4/114) in RPUL group and 17.4％ (16/92) in URL group.The stone-free rate was 100.0％ (114/114) in RPUL group and 89.1％ (82/92) in URL group.The differences were significant (P ＜ 0.05).Conclusions RPUL and URL had the advantages of less trauma and blood loss and rapid recovery.RPUL had fewer complication and higher success rate than URL,and could be a minimally invasive option for the treatment of ureteral calculi.

Objective To develop a digital polymerase chain reaction (PCR) ribotyping method and database for Clostridium difficile genotyping.Methods Sequencer based fluorescence capillary gel electrophoresis was used,instead of agarose gel electrophoresis,to establish the digital PCR-ribotyping of Clostridium difficile.Forty Clostridium difficile reference strains,consisting of 10 PCR-ribotypes (RT),were genotyped by the new digital PCR-ribotyping method to set-up the database.Results The sequencer based fluorescence capillary gel electrophoresis correctly detected PCR-ribotyping products of the 40 reference strains,and showed as digital figure;significant differences of these digital figures were found between the 10 RT.High similar digital figures were shown in twenty-one RT027 strains,three RT002 strains and two RT014 strains.However,seven RT001 strains were typed as four subtypes,and two RT014 strains as two subtypes,respectively.Conclusion A digital PCR-ribotyping,and a reference database consisting of 10 RT are successfully established.

Objective To investigate the genotype and production of toxin A and B of C .difficile clinical isolates collected from Sydney ,Australia .Methods Sixty‐eight C .difficile clinical isolates were collected from Westmead Hospital ,the University of Sydney ,which were genotyped by using PCR‐ribotyping ,and toxin A ,B coding gene tcdA ,tcdB were detected by using PCR meth‐od .Results Thirty‐one PCR‐ribotypes (RTs) were confirmed in the 68 C .difficile clinical isolates ,RT014 (19 .1% ) and RT002 (11 .8% ) were the common genotypes .Sixty‐four of 68 (94 .1% ) isolates contained tcdA and tcdB for toxin A and B .Conclusion The common prevalent PCR‐ribotypes of C .difficile were RT014 and RT002 in Sydney ,most of the C .difficile clinical isolates contained toxin A and B .

Objective To investigate the genotype and variance of toxin associated genes of moxifloxacin‐resistant Clostridium difficile clinical isolates in Sydney .Methods Twenty‐two moxifloxacin‐resistant Clostridium difficile clinical isolates were collected from Sydney ,which were genotyped by using sequencer capillary gel electrophoresis based PCR‐ribotyping ,and toxin A and B cod‐ing gene tcdA and tcdB ,and binary toxin coding gene cdtA and cdtB were detected by using PCR method .Toxin regulator gene tc‐dC was analyzed by using PCR‐sequencing ,and was aligned with reference sequence of VPI 10463 (Genbank accession number :X92982) ,and the tcdC sequence types of all 22 isolates were identified by using blast tool in NCBI .Results Twenty‐one isolates were genotyped as hypervirulent PCR‐ribotypes 027 (RT027) ,and one isolate as RT078 ;all 22 isolates contained tcdA and tcdB for toxin A and B and cdtA and cdtB for binary toxin (tcdA+ tcdB+ cdtA+ cdtB+ ) .The tcdC sequence types of the 21 RT027 i‐solates belong to sc1 ,and that of the one RT078 isolate belongs to WA39 .Compared with tcdC reference sequence of VPI 10463 ,a consecutive 18 bp deletion (nt341 to 379) and one nucleotide deletion at position 117 were found in the 21 RT027 isolates ,and a consecutive 39 bp deletion (nt330 to 368) and one nucleotide mutation at position 184(C> T) were found in the one RT078 isolate . Conclusion Clostridium difficile hypervirulent RT027 was the common moxifloxacin resistant genotype ;Clostridium difficile hy‐pervirulent RT027 and RT078 clinical isolates contained genes for toxin A and B and binary toxin ,and contained gene sequence mu‐tation in toxin regulator gene tcdC .

BACKGROUND:Whether long time exposure to the lead concentration which is withinthe state allowed range will cause any bad effects on people's health?OBJECTIVE: To investigate the nerve conducting velocity (NCV) and other biological indices of workers who are exposed to low concentration lead.DESIGN: It was an investigation and the subjects were workers exposed to low concentration lead.SETTING: Occupational Disease Department and Electrophysiological Department of Xinhua Affiliated Hospital of the Shanghai Second Medical University.PARTICIPANTS: In lead contact group were 66 heat treatment workers who were exposed to lead and received physical examination from September 2000 to October 2002. In control group were 40 office workers that worked at the same factory but were not exposed to lead.METHODS: Questionnaire and laboratory examination were adopted for measurement of NCV and other biological indices like blood lead, blood zinc protoaetioporphyrin (ZPP), blood free protoporphyrin (FPP) and hemoglobin content in the two groups. And the relative risk analysis was conducted.biological indices in both groups.RESULTS: All the 106 subjects entered analysis stage. In lead contact group there were 34 workers of more than 10 working years and the other ulnar nerve in those of over 10 working years were faster than those in control group [(50.11 ±4.76) m/s, (63.11±2.58) m/s vs (47.59±4.86)m/s,those of over 10 working years was higher than those of less than 10years [(0.568±0.28), (0.425±0.31) μmol/ L, P ＜ 0.05]. So was the FPP level [(2.24±0.32), (2.09±0.27) μmol/L, P ＜ 0.05] and urine lead level [(0.087±0.008), (0.083±0.007) μmg/L, P ＜ 0.05]. The ZPP level of those of over 10 working years was also higher than those less than 10 years [(1.42±0.33) μmol/L vs (1.25±0.35) μmol/L, P ＜ 0.05] and control [(1.42±0.33) μmol/L vs (1.22±0.44) μmol/L, P＜ 0.05]. The hemoglobin of those of over 10 working years was lower than those of less than 10 years [(12.26±4.5) g/L vs (14.55±4.81) μmol/L, P ＜ 0.05] and control [(12.26±4.5) g/L years presented abnormality in conducting sensory signals and the abnormality rate was 3% (1/32). Whereas four cases in those with more than 10working years did so and the abnormality rate was 12% (4/34). As for biological indices, one case in those less than 10 working years was beyond normal values (abnormality rate 3%) and five did so in those of over 10working years (abnormality rate 15%). It could be seen that changes in both NCV and biological indices of those of over 10 working years were more serious than those les than 10 years exposure with the relative risks of 4.1 and 5.3 respectively.CONCLUSION: Persistent exposure to lead not only alters biological indices but also damages nervous system in different degrees. Sensory verve of lower extremities and motor nerves of upper extremities are usually damaged first. Peripheral nerve injury is common in workers of over 10 working years. Because the regeneration of nervous system is poor, so the nerve injury is usually irreversible. Therefore the neuroelectrophysiological changes always predict early nerve injury and so neuroelectrophysiological monitoring can be adopted for prevention of nerve injury.

The genetic divergence between geographically separated populations can be studied by comparing the divergences in their allele frequencies to the geographical distance of popula-tions. The present study is to examine the divergences in allele frequencies and genetic diver-gence in relation to the geogrephical distance of the samples localities, in order to test the population structure model of Oncomelania from the endemic areas in mainland China.The results showed that three patterns of allele frequency distributions occured in seven polymorphic loci, which were even,chaotic and discontinuous cline distributions. The distcon-tinuous or stepped clines shown in loci Est-4,Got and Mdh-2 suggested that the discrete subpopuation model is the likeliest. The plot of genetic distance (Nei's 1978) between sample populations against the geographic distances suggested that the special pattern of allele fre-quency distribution could be found. The regression analysis shows that logistic sigmoid re-gression is the best model to fit the original data in the plot. This supports the existence of the discrete subpopulations model in the population structure studied.

BACKGROUND:Cytokine induced kil er cells therapy as an effective means of adoptive immunotherapy, becomes a new way to treat acute myeloid leukemia. But, the researches about sequential cytokine induced kil er cells therapy after autologous peripheral blood stem celltransplantation in acute myeloid leukemia patients are stil less, which deserve further research.
OBJECTIVE:To observe the clinical efficiency and safety of sequential cytokine induced kil er cells therapy after autologous peripheral blood stem celltransplantation in acute myeloid leukemia M2 patients.
METHODS:Total y 45 patients with low-or intermediate-risk acute myeloid leukemia M2 were recruited in this study. Among them, 19 patients received sequential cytokine induced kil er cells therapy after autologous peripheral blood stem celltransplantation and 26 patients only received autologous peripheral blood stem celltransplantation. The relapse rate, disease-free survival, and overal survival were compared between two groups, and safety of cytokine induced kil er cells therapy was observed.
RESULTS AND CONCLUSION:(1) Compared with the patients only receiving autologous peripheral blood stem celltransplantation, the relapse rate was lower (21.05%vs. 38.46%;P<0.05), and elevated percentages of the disease-free survival and overal survival were observed in the patients receiving sequential cytokine induced kil er cells therapy after autologous peripheral blood stem celltransplantation (P<0.05). (2) The 19 patients who received sequential cytokine induced kil er cells therapy after autologous peripheral blood stem celltransplantation al completed the treatment scheme successful y. Only four patients appeared to have chil s and fever, and no more side effects were observed. These findings suggested that the sequential cytokine induced kil er cells therapy after autologous peripheral blood stem celltransplantation can improve the disease-free survival and overal survival of low-or intermediate-risk acute myeloid leukemia M2 patients without remarkable side effects, which is a safe, effective and feasible way for the treatment of acute myeloid leukemia M2.

Objective To investigate the carrying status and characteristics of Clostridium difficile isolated from infants.Methods Two hundred and thirty-eight stool specimens were collected from infant younger than 1 year old,that were hospitalized or outpatient from August to November 2015.Immunochromatography targeted GDH and toxin A&B of C.difficile was used for C.difficile screening,and those positive specimens were inoculated in CDIF and anaerobic culture.C.difficile isolates were genotyped by using slpA sequence typing (slpA ST),and tcdA,tcdB,cdtA and cdtB of C.difficile isolates were detected by PCR.Results Fifty C.difficile strains were isolated from 238 stool samples,and the isolated rates of C.difficile from <3 months,3 months to <6 months,and 6 months to 1 years old groups were 9.3%,17.6% and 27.3%(χ2=6.940,P=0.031<0.05),respectively.52.0%(26/50) of the C.difficile isolates were toxigenic,and 69.2% (18/26) toxigenic isolates harbored tcdA+tcdB+cdtA-cdtB-.Fifty C.difficile isolates were genotyped as 11 slpA STs,slpA ST fr-02 and kr-02 were the commonest genotypes in toxigenic C.difficile isolates;however,that was slpA ST xr-03 in non-toxigenic isolates.Conclusion High C.difficile carriage is found in infants younger than 1 year old,and more than half of C.difficile isolates are toxigenic.Most of toxigenic isolates harbored toxin A and B.The genotype of C.difficile isolates is different between toxigenic isolates and non-toxigenic isolates.