Objective To prepare a monoclonal antibody(McAb) against Trichinella spiralis (T.s) adult and study its protective immunity. Methods BALB/c mice were immunized with soluble antigens of 3 day adult of T.s. Immnuized spleen cells of BALB/c mice were fused with myeloma cells sp2/0. The hybridoma culture supernatant was isotyped by the Ouchterlony double diffusion technique and the specificity of the McAb was determined by immunoblotting. The protective immunity of the McAb was detected after challenging the mice by the intravenous way. Results A McAb against T.s adult was obtained. The titer of culture fluid and ascetic fluid was 1∶6 400 and 1∶204 800, respectively. This McAb was identified as IgM. Western blotting results showed this McAb can be used to identify 40 000 protein of adult worm, muscle larvae and newborn larvae of T.s. No cross reactions with antigens of Ascaris suum Goeze, Taenia solium Linnaeus, Echinococcus granulosus and Schistosoma were oberserved. The number of muscle larvae was decreased by 55.63% when giving McAb to mice intravenously. Conclusions A species specific McAb against T.s adult was established and its protective immunity was identified. This McAb can be used as a powerful tool in screening Trichinella vaccine candidates.

Objective To detect circulating antigen (Cag) for diagnosing neurocysticercosis. Method ELISA was performed with monoclonal antibody 4B6 against the cyst fluid antigen of Cyticercus cellulosae for detecting Cag in serum and/or cerebrospinal fluid (CSF) of patients with neurocysticercosis or with other diseases. Results In the group of 82 cases of neurocysticercosis, the positive rate of serum Cag was 79.2% (65/82) and the positive rate of CSF Cag was 100% (26/26). After chemotherapy for 20 cases with positive serum Cag, the titer of serum Cag in 17 cases dropped to zero(85% ). Cag could not be detected in specimens from patients with other diseases. Conclusion These results indicate that the determination of Cag, especially of the CSF Cag, is useful for the diagnosis of neurocysticercosis and the drop in serum Cag is a good parameter for the evaluation of the effectiveness of chemotherapy.

Objective To establish axenic cultivation of Pneumocystis carinii (P.c). Methods The organisms of P.c were isolated from the bronchoalveolar lavage fluid (BALF) of the rats with Pneumocystis carinii pneumonia (PCP) and cultured in a medium which was based on IMDM (GIBCO) supplemented with S-adenosyl-L-methionine, putrescine, N-acetyl glucosamine, putrescine, L-cysteine and L-glutamine, and newborn calf serum. The organisms cultured in the system were identified by observing the morphology of cysts in smears stained with Gomori's methenamine silver nitrate stain (GMS). Ultrastructure of the cysts/trophozoites was examined by transmission electron microscopy. The sequences of mitochondria] large ribosomal DNA subunit of the cutured organisms were compared with the Pneumocysti carinii f.sp. ratti variant isolate (GenBank No U20173) and Pneumocystis carinii f.sp.hominis (GenBank No M58605). Results Five isolates of P.carinii received from BALF of 8 rats with PCP were cultured axenically and continuously in the system. The cultured organisms could be stored in frozen condition and used to reinitiate culture, and were amplified by 19-22 times within 72 h. The morphology, ultrastructure and gene sequencing of the cultured organisms confirmed that the isolated organisms were P.carinii. Conclusion Five continuously and axenicly cultured isolates of P.carinii have been received.

Objective To investigate the effects of mefformin on the differentiation of osteoclastas well as relative mechanism.Methods Raw264.7 cells from the murine macrophage cell line was used.Receptor activator of NF-κB ligand (RANKL) was used to stimulate osteoclast differentiation from Raw264.7 cells.Osteoclast differentiation was assessed by tartrate-resistant acid phosphatase (TRAP) and actin fluorescence staining and counting the TRAP-positive cells after exposure to different concentrations of mefformin (0 μmol/L,400 μmol/L,800 μmol/L and 1000 μmol/L) or rapamicin (100 nmol/L) in the presence of 50 ng/ml RANKL for 5 days.Bone-resorbing activity was evaluated by BD BioCoatTM OsteologicTM Bone Cell Culture System.The expression of osteoclast-specific genes like TRAP,capthesin K,calcitonin receptor (CTR) and matrix metalloproteinase (MMP-9) was evaluated by RT-PCR.The expression of tumor necrosis factor-α(TNF-ct) S6K1Thr389,S6 Ser235/236,4E-BP1Thr37/46 and c-Fos protein was evaluated by ELISA kit and Western blot analysis,respectively.Results Mefformin dose-dependently inhibited RANKL-stimulated osteoclasts differentiation in Raw264.7 cell culture,as manifested by decrease of TRAP-positive multinucleated cells and pit erosion area,down-regulation of TRAP,cathepsin K,CTR and MMP-9 mRNA and reduction of TNF-α and c-Fos protein expression.Further study revealed that RANKL activated mTOR complex 1(mTORC1) signaling,while mefformin impaired RANKL-stimulated mTORC1 signaling.Rapamycin,an mTORCl-specific inhibitor and immunosuppressive macrolides could also prevent RANKL-induced osteoclast differentiation and bone resorption in vitro.Conclusion Mefformin inhibits osteoclastogenesis in vitro,which may due to reduction of TNF-α and c-Fos protein expression,and mTORC1 signaling is involved in this process.

Objective To analyze the clinical features of 6 patients with imported schistosomiasis mansoni,including the epidemic history,clinical manifestations,laboratory tests and therapeutic effect,so as to provide references for improving the levels of diagnosis and treatment of physicians. Methods The clinical data of 6 patients with imported schistosomiasis mansoni from January 2009 to July 2016 were collected and analyzed. Results All the 6 imported patients with schistosomiasis mansoni had a clear history of cercarial infested water exposure. The main manifestations were continuous fever and eosinophilia. Three (50%)patients were accompanied with diarrhea. Anti-Schistosoma japonicum IgG antibody were cross positive in 2(33.3%)pa-tients,while live eggs of S. mansoni were explored in intestinal mucosa specimens of all the patients. CD3+CD8+T cell ratio was decreased significantly but B cell ratio was elevated in all the patients,and the main immunoglobulin of the patients was IgG. Hydroperitoneum and splenomegaly signs were discovered by abdominal ultrasonography in 16.6%(1/6)of the patients. Multi-ple liver nodules and wall thickening of rectum and sigmoid colon were revealed by pelvic MR scan in 16.6%(1/6)of the pa-tients. Colitis was found in all the patients,and 66.6%(4/6)of the patients were combined with multiple colonic ulcers by the electronic colonoscopy examination. Chronic inflammation and eosinophil infiltration were found in all the patients by rectum pa-thology. All 6 patients were cured with chemotherapy named praziquantel. Conclusion Comprehensive analysis of clinical data including epidemiological history,specific manifestations,laboratory tests and intestinal mucosa pathology may be benefit of the management of schistosomiasis mansoni.

Objective To investigate antimicrobial resistance among gram-negative bacilli in China in 2010.Methods A total of 1 259 consecutive and non-repetitive gram-negative bacilli were isolated from 13 teaching hospitals from September to December in 2010 in China.All of these isolates were sent to the central laboratory for re-identification and susceptibility testing.The MIC of meropenem and other antibacterial agents were determined by agar dilution method.Interpretive results was determined by CLSI M100-S21.Results The activity of 14 antibacterial agents against 845 Enterobacteriaceae isolates was as follows in order:meropenem ( 98.1％,829 ),amikacin ( 94.0％,794 ),imipeuem ( 90.0％,761 ),piperacillin/tazobactam ( 87.5％,739 ),cefepime ( 83.0％,701 ),ertapenem ( 82.4％,696 ),cefoperazone/sulbactam ( 80.3％,678 ),colistin (75.4％,637),ceftazidime (70.0％,591 ),ciprofloxacin (59.1％,499 ),cefoxitin ( 54.8％,463 ),ceftriaxone ( 53.5％,452 ),cefotaxime ( 52.3％,442 ) and minocycline(51.5％,435).The prevalence of ESBL was 61.3％ (106/173) in Escherichia coli,which was much higher than 41.2％ (70/170) in Klebsiella pneumoniae.The susceptibility rate of Klebsiella pneumoniae against meropenem,imipenem,amikacin and colistin were more than 90％,but were highly resistant to ceftriaxone and cefotaxime.Over 80％ of Enterobacter cloacae,Enterobacter aerogenes,Citrobacter freundii,were susceptible to meropenem,amikacin,cefepine,cefoperazone/sulbactam,imipenem,piperacillin/ tazobactam,and ertapenem.The most active antibiotics against Pseudomonas aeruginosa were Colistin (98.4％,182),Amikacin ( 85.9％,159 ),Piperacillin/Tazobactam ( 80％,148 ),Ceftazidime ( 79.5％,147),Meropenem (74.1％,137),Ciprofloxacin (74.1％,137),Cefepime (73.5％,136),Imipenem (71.9％,132) and Cefoperazone/Sulbactam (70.8％,131 ).Less than 37％ of Acinetobacter baumannii isolates were resistant to carbapenems.The susceptible rate to Minocycline was 47.8％.Colistin kept good activity against Acinetobacter baumannii (susceptible rate,97.8％,n =176),The prevalence of Pan-drug resistant P.aeruginosa and A.baumannii was 18.9％ (n =35),and 61.8％ (n =108),respectively.Conclusions Carbapenems remained very high activity against Enterobacteriaceae.Increasing resistance to the antimicrobials agents test among A.baumanni and P.aeruginosa,especially carbapenems among A.baumanni brought great concern.

Objective To investigate the clinical results of selective decompression and short-segment fusion with fixation for symptomatic degenerative lumbar stenosis combined with lumbar scoliosis.Methods All of 30 patients from Jul.2008 to Oct.2013 were recruited for this retrospective study.There were 11 males and 19 females,whose mean age was 60.3±12.7 years.The preoperative X-ray of the total spine showed the mean Cobb's angle was 24.3°±8.8°.And the mean lumbar lordosis angle was 30.5°±15.5°.Pain and function were assessed by Visual Analogue Scale (VAS) and Oswestry dsability index (ODI).The responsible segments were determined from physical examination and radiological findings.Selective decompression and short-segment fixation and fusion were performed.The radiographic parameters,ODI,VAS of pre-operation and post-operation were recorded and compared.Results All the patients were followed up for 21-73 months with mean 46.0±10.9 months.The complication incidence was 33.3％.The ODI and VAS assessment was significantly improved during the follow-up,as well as the sagittal and coronal radiographic parameters (LL,SS,PT,SVA,Cobb's angle,C7PL-CSVL).The improvement of VAS and ODI of lumbar spine was significant correlated with sagittal parameters (LL,PT),whilst not correlated with coronal parameters.Conclusion The surgical strategy of selective decompression and short-segment fusion with fixation is effective for the patients with symptomatic degenerative lumbar stenosis combined with lumbar scoliosis.

Objective To evaluate the performance of modified Hodge test on the detection of carbapenemases among Enterobacteriaceae with decreased susceptibility to carbapenems. Methods Fortynine Enterobacteriaceae isolates with decreased susceptibility to carbapenems ( MIC of imipenem, meropenem or ertapenem was ≥ 2 μg/ml ) were collected from 16 teaching hospitals from 2004 to 2008. MICs of imipenem, meropenem and etapenem were determined by agar dilution method. Carbapenemases were detected by modified Hodge test. Carbepenemase-causing positive results and AmpCs-causing positive results were differentiated by phenyl boronic acid and oxacillin. Beta-lactamases encoding genes including blaNDM-1were detected by PCR and sequencing. Results Thirty-six of 49 isolates were non-susceptible to imipenem (MIC >4 μg/ml), 31 were non-susceptible to meropenem (MIC > 4 μg/ml) and 47 were non-susceptible to ertapenem (MIC > 2 μg/ml). Twenty-three isolates showed positive modified Hodge test result, including 9 weak-positive results and 14 strong-positive results. Through PCR detection and sequencing, 2 out of 9 isolates showing weak-positive results carried blaKPC-2 and other 7 did not carry any carbapenemase genes but AmpCs/ESBLs genes. Among the 14 isolates showing strong-positive results, 4 carried blaKPC-2, 8 carried blaIMP-4 and 2 caried blaIMP-8. All 26 isolates with negative modified Hodge test result didn't carry any carbapenemase genes. No isolate carried blaNDM-1. Carbapenemases genes PCR detection was regarded as a gold standard, and the sensitivity, specificity, positive predictive value and negative predictive value of modified hodge test was 100%, 79%, 70% and 100% on the detection of carbapenemases among Enterobacteriaceae with decreased susceptibility to carbapenems. Conclusions Modified Hodge test revealed great sensitivity but showed a few false positive results. True and false positive results can be effectively differentiated by phynel boronic acid and oxacillin.

80% activity rate against E.coli included piperacillin/tazobactam(93.4%)、ceftazidime(86%),and amikacin(83.3%);The susceptible rate to piperacillin/tazobactam in K.pneumoniae was 84.6%. The susceptible rate to ceftazidime decreased from 82.3% to 69.9%, which was lower than to cefepime (77.2%). Over 50% of Enterobacter cloacae were resistant to ceftazidime, cefotaxime and ceftriaxone. Susceptible rates to piperacillin/tazobactam in E. cloacae,E. aerogenes,Citrobacter freundii and Serratia marcescens (67.7%-96.4%) were higher than those to cefepime (68.8%-77.5%), cefoperazone/sulbactam (59.7%-87.5%). Susceptibility to amikacin among these 4 species (70%-83.7%) was higher than to ciprofloxacin (48.1%-79.5%). All of Morganella morganii and Proteus vulgaris isolates were susceptible to meropenem and imipenem; Over 90% of the isolates were susceptible to cefepime, cefoperazone/sulbactam and piperacillin/tazobactam.The most active agent against Pseudomonas aeruginosa was meropenem (84%), followed by amikacin, piperacillin/tazobactam, ceftazidime and imipenem (72.5%-76.6%). Mutiple-drug-resistant Acinetobacter baumannii increased from 33% in 2003 to 48% in 2004. Resistance to carbapenems increased to 18% in this species in 2004. The most active agents against Burkholderia cepacia were meropenme (64.9%), cefoperazon/sulbactam (63.2%), ceftazidime (59.6%), piperacillin/tazobactam (56.1%) and cefepime (52.6%).Conclusions Carbapenems remained very high activity against Enterobacteriaceae. Increasing resistance to 10 antimicrobials agents tested among A. baumanni brought great concern. Meropenem was 4-to 16-fold more active against common gram-negative bacilli than imipenem.

With the increasing proportion of human papilloma virus(HPV)infection in the pathogenic factors of oro-pharyngeal cancer,a series of changes have occurred in the surgical treatment.While the treatment mode has been im-proved,there are still many problems,including the inconsistency between diagnosis and treatment modes,the lack of popularization of reconstruction technology,the imperfect post-treatment rehabilitation system,and the lack of effective preventive measures.Especially in terms of treatment mode for early oropharyngeal cancer,there is no unified conclu-sion whether it is surgery alone or radiotherapy alone,and whether robotic minimally invasive surgery has better func-tional protection than radiotherapy.For advanced oropharyngeal cancer,there is greater controversy over the treatment mode.It is still unclear whether to adopt a non-surgical treatment mode of synchronous chemoradiotherapy or induction chemotherapy combined with synchronous chemoradiotherapy,or a treatment mode of surgery combined with postopera-tive chemoradiotherapy.In order to standardize the surgical treatment of oropharyngeal cancer in China and clarify the indications for surgical treatment of oropharyngeal cancer,this expert consensus,based on the characteristics and treat-ment status of oropharyngeal cancer in China and combined with the international latest theories and practices,forms consensus opinions in multiple aspects of preoperative evaluation,surgical indication determination,primary tumor re-section,neck lymph node dissection,postoperative defect repair,postoperative complication management prognosis and follow-up of oropharyngeal cancer patients.The key points include:① Before the treatment of oropharyngeal cancer,the expression of P16 protein should be detected to clarify HPV status;② Perform enhanced magnetic resonance imaging of the maxillofacial region before surgery to evaluate the invasion of oropharyngeal cancer and guide precise surgical resec-tion of oropharyngeal cancer.Evaluating mouth opening and airway status is crucial for surgical approach decisions and postoperative risk prediction;③ For oropharyngeal cancer patients who have to undergo major surgery and cannot eat for one to two months,it is recommended to undergo percutaneous endoscopic gastrostomy before surgery to effectively improve their nutritional intake during treatment;④ Early-stage oropharyngeal cancer patients may opt for either sur-gery alone or radiation therapy alone.For intermediate and advanced stages,HPV-related oropharyngeal cancer general-ly prioritizes radiation therapy,with concurrent chemotherapy considered based on tumor staging.Surgical treatment is recommended as the first choice for HPV unrelated oropharyngeal squamous cell carcinoma(including primary and re-current)and recurrent HPV related oropharyngeal squamous cell carcinoma after radiotherapy and chemotherapy;⑤ For primary exogenous T1-2 oropharyngeal cancer,direct surgery through the oral approach or da Vinci robotic sur-gery is preferred.For T3-4 patients with advanced oropharyngeal cancer,it is recommended to use temporary mandibu-lectomy approach and lateral pharyngotomy approach for surgery as appropriate;⑥ For cT1-2N0 oropharyngeal cancer patients with tumor invasion depth＞3 mm and cT3-4N0 HPV unrelated oropharyngeal cancer patients,selective neck dissection of levels ⅠB to Ⅳ is recommended.For cN+HPV unrelated oropharyngeal cancer patients,therapeutic neck dissection in regions Ⅰ-Ⅴ is advised;⑦ If PET-CT scan at 12 or more weeks after completion of radiation shows intense FDG uptake in any node,or imaging suggests continuous enlargement of lymph nodes,the patient should undergo neck dissection;⑧ For patients with suspected extracapsular invasion preoperatively,lymph node dissection should include removal of surrounding muscle and adipose connective tissue;⑨ The reconstruction of oropharyngeal cancer defects should follow the principle of reconstruction steps,with priority given to adjacent flaps,followed by distal pedicled flaps,and finally free flaps.The anterolateral thigh flap with abundant tissue can be used as the preferred flap for large-scale postoperative defects.