Objective To investigate the cross reaction characteristics of recombinant human papillomavirus 16 type L2 full-length fusion protein in HPV types of 6, 11, 18.Methods The serum samples of 108 condyloma acuminatum patients, 156 cervix cancer patients and 100 healthy control subjects were collected.The gene of full-length HPV16 L2 was amplificated from the tissue DNA of cervical cancer patient and inserted into the prokaryotic expression vector PGEX-4T-1 to construct the recombinant plasmid PGEX-4T-1-HPV16 L2.After sequencing identification, the recombinant plasmid was tranformed into E.coli BL21 (DE3).After induction by IPTG, the fusion protein containing HPV16 L2 was expressed and analyzed by both SDS-PAGE and WB.Furthermore, the specific binding capacity of the fusion protein to the HPV 6,11, 16 and 18 DNA positive patient's sera were analyzed by WB.The fusion protein was purified with NiNTA Agarose Kit and coated with ELISA reaction plates.The specific serum IgG were detected by indirect ELISA.Results The recombinant plasmid PGEX-4T-1-HPV16 L2 was constructed successfully. Highly expressed HPV16 L2 full-length fusion protein was obtained and the expression level was 27.2 %.The relative molecular mass(Mr) of the fusion protein is about 82 × 103, which matches up to the expected Mr.Meanwhile, the sera of HPV 6,11,16,18 DNA positive patients were used as the primary antibody and the Mr of the specific band was detected to be about 82 × 103 by WB.The results of indirect ELISA showed that the average levels of specific IgG in condyloma acuminatum group, cervical cancer group and healthy control subjects were 0.848 ±0.257, 0.822 ±0.247 and 0.173 ±0.143 with the positive rate of 92.6%, 94.2%and 8.0% respectively.There was no significance of the specific IgG levels between condyloma acuminatum group and cervical cancer group ( F = 0, P ＞ 0.05 ), but there was significant difference of specific IgG levels and positivity among the three groups ( F = 305.201 ,x2 = 253.178, P ＜ 0.01 ).Conclusions The HPV16 L2 full-length fusion protein has better antigenicity.However cross reactions with HPV6, 11 and 18 were found.It can be applied in serological screening reagents for HPV infection and associated cancer.

Objective To analyze the drug resistance gene carrying situation of carbapenem-resistant Klebsiella pneumoniae (CRKP) and molecular epidemiological characteristics to provide a reference basis for studying the bacterial drug resistance.Methods A total of 37 clinically isolated strains of CRKP were collect ed from February 2016 to February 2017.The broth microdilution method was used to determine the strain drug susceptibility.The phenotypes of CRKP carbapenemases were detected by using the modified Hodge test and EDTA-imipenem synergistic method.The drug resistance genes of KPC-2,NDM-1 and OXA-48 were detected by PCR.The sequencing and internet comparison were performed for determining the genotype.The multilocus sequence typing (MLST) was adopted to conduct the genetic correlation study on the strains.The evolutionary trees were constructed by using the MEGA software and the genetic relationship was analyzed by using the eBURST software.Results The drug resistance to commonly used antibacterial drugs was over 90％.The KPC-2 gene was detected in all strains,3 strains simultaneously carried the NDM-1 gene,and other genes were negative.In MLST typing,25 strains were ST11,each 2 strains were ST524 and ST789,each 1 strain was ST35,ST29,ST1066 and ST244 respectively.Also a new ST type(2 strians) was confirmed by the PubMlst database and named as ST1792.The ST11 type group and non-ST11 type group had no statistical difference in the aspects of the age,sex,infection route and antibiotics use(P＜0.05).Conclusion Carrying KPC-2 gene is the main cause leading to bacterial resistance to carbapenem and ST11 type is the most popular clone type.

OBJECTIVETo assess the feasibility of using saliva for Schistosomiasis japonica diagnosis.

METHODSSchistosoma japonicum infected animal model was established. Pairs of saliva and serum samples from rabbits and chronic schistosomiasis patients were collected. Anti-schistosoma specific antibodies in saliva and serum were detected by indirect ELISA.

RESULTSThe specificities of antibody detection of rabbit saliva and serum were 93% (28/30) and 97% (29/30), respectively, and the sensitivities of antibody detection of rabbit serum and saliva were 100% (24/24) and 88% (21/24), respectively. A significant correlation (r = 0.5307, P = 0.0038 < 0.05) existed between anti-SEA IgG levels in serum and saliva. As with those in serum, anti-SEA IgG levels in saliva could reflect the state of infection and treatment. The sensitivity of antibody detection was 91% (29/32) for patient saliva samples and 100% (32/32) for their sera. 8 samples were positive in 140 normal saliva samples (i.e. 6% false positive rate) and 6 samples were positive in 156 normal serum samples (4% false positive rate). There was a significant correlation (r = 0.4227, P = 0.008 < 0.05) between specific antibodies in saliva and serum.

CONCLUSIONThe detection of specific antibodies in saliva can be used as a non-invasive immunodiagnosis method of Schistosomiasis japonica.

Adolescent ; Adult ; Animals ; Antibodies, Helminth ; analysis ; Child ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Immunoglobulin G ; analysis ; Male ; Middle Aged ; Rabbits ; Saliva ; immunology ; Schistosoma japonicum ; immunology ; Schistosomiasis japonica ; diagnosis

Objective To evaluate the effect of electroacupuncture at Chize and Shangjuxu on the prognosis and respiratory dynamics in patients with acute respiratory distress syndrome(ARDS).Methods A prospective randomized controlled study was conducted to collect patients with ARDS admitted to the Department of Emergency Medicine of Tongde Hospital of Zhejiang Province from December 2020 to September 2022.A total of 142 patients with ARDS from the Department of Emergency Medicine of Tongde Hospital of Zhejiang Province were selected for this study.Seven patients were excluded due to rapid deterioration,3 patients withdrew the informed consent,and finally 132 patients were enrolled.According to the order of enrollment,they were randomly divided into the electroacupuncture group and the conventional treatment group based on the random number table.The conventional treatment group received conventional treatment for ARDS,while the electroacupuncture group combined with electroacupuncture at Chize and Shangjuxu,the treatment lasted for 30 minutes.The mortality at day 60 and secondary efficacy indicators such as hospital stay,ventilator-free days at 28 day,and mechanical ventilation time of two groups were compared,and the indicators of respiratory mechanics and oxygenation such as inhalation oxygen concentration(FiO2),tidal volume(VT),positive end-expiratory pressure(PEEP),plateau pressure(Pplat),lung static compliance(Cst),oxygenation index(PaO2/FiO2)were also compared.Then the Kaplan-Meier survival curves were drawn to compare the differences in cumulative survival rates within 60 days between the two groups.Results The mortality at day 60 in the electroacupuncture group was lower than that in the conventional treatment group[33.33％(22/66)vs.40.91％(27/66)],but the difference was not statistically significant(P>0.05),and the hospital stay in the electroacupuncture group was significantly shorter than that in the conventional treatment group(days:25.09±11.57 vs.30.21±15.94,P<0.05).The ventilator-free days at 28 days in the electroacupuncture group were significantly higher than those in the conventional treatment group(days:10.06±7.35 vs.7.67±5.72,P<0.05).There was no statistically significant difference in mechanical ventilation time between the two groups(days:15.70±9.05 vs.18.39±9.55,P>0.05).After 72 hours of treatment,the FiO2,VT,and PaCO2 levels in the electroacupuncture group were significantly lower than those in the conventional treatment group[FiO2:0.39±0.07 vs.0.44±0.09,VT(mL/kg):6.27±0.74 vs.6.62±0.74,PaCO2(mmHg,1 mmHg≈0.133 kPa):39.94±4.52 vs.41.95±4.19,all P<0.05].Conclusion Electroacupuncture at Chize and Shangjuxu can effectively shorten the course of ARDS and improve prognosis,which may be related to the improvement of respiratory dynamics by electroacupuncture treatment.

With the development of modern medical technology,accurate resection of tumor and timely repair and repair of defective tissues and organs are important concerns in the field of tumor research.The precise excision of tumor,refers to the preoperative assessment of systemic and local detection based on detailed to personalized surgical planning,the use of precise operation in operation,ensure as much as possible while minimizing surgical trauma to patients after removal of the lesions,creating the optimal conditions of recovery for trauma patients.Repair and regeneration of defective tissues and organs refers to the deletion or damage of tissues and organs,and gradually resume its anatomical structure and function process under the action of a variety of cells,extracellular mechanisms and related regulatory factors.Then from the tumor resection,tumor resection and accurate regeneration after three point repair technology to change rapidly in the tissue of tumor plastic organ regeneration in tissue of origin.