In combination with the physiologic classroom instruction of high altitude pathophysiology,this text discusses ten kinds of heuristic teaching methods.These methods have certain functions of relaxing the atmosphere of the classroom,encouraging students'enthusiasm and helping to formulate their thinking ability.But we shoud grasp three principles in carrying out the heuristic education.First,the inspiration should have pertinency.Second,we should consider the accepting level of the students.Third,we should make best use of the situation and advance step by step.This article has certain directive significance to the classroom instruction.

The spleen cells from the BALB/C mouse immunized with the type B toxoid were fused with aNS-1 myeloma cell line. The superatant containing the hybridoma-formed cells with growing pores wasscreened by EL1SA, in which 66.7% the pores could secrete the specific antibodies against botulin.After a Subclonizing culture by the limiting dilution technique four hybridoma cell lines(3B10, 3B11, 3G12 and 4A5) were established and could secrete the specific antibodies persistently in the culture medium. in which antibody titers came to 10-3-10-5, while they were injected into the BALB/C mice intrape ritoneally ascites rich in antibodies with a titer of 10-5-10-8 was produced. The results testing the (our monoclonal antibodies with the type A and B toxoids showed that the antibodies of 3G12and 4A5 were specific for the type B toxoid, and those of 3B10 and 3B11 had light cross reaction with the type A toxoid. Identification of 1g showed that the antibodies of 3B10 and 3G12 were of IgG1, while those of 3B11 and 4A5 of IgG2. The chromosomal assay confirmed the four cell lines to be hybridoma. The neutroligation test in mice revealed that those four monoclonal antibodies did not show any protective effects on botulin.

[ ABSTRACT] AIM: To investigate the seroprevalence of neutralizing antibodies to human adenovirus type 5 (AdHu5) , human adenovirus type 26 (AdHu26) and chimpanzee adenovirus type 68 (AdC68) in the patients with chro-nic hepatitis B ( CHB) and the patients with primary liver cancer ( PLC) , and to provide guidance for developing safe and effective biotherapy vectors against CHB and PLC.METHODS:The blood samples from 196 patients with CHB and 193 patients with PLC were examined to assess the presence of neutralizing antibodies against AdHu5, AdHu26 and AdC68 by adenovirus neutralization assays.RESULTS:The seroprevalence rates of neutralizing antibodies to AdHu5, AdHu26 and AdC68 in the CHB patients were 84.7%, 58.2%and 39.8%, respectively.Among the patients with PLC, the prevalence rates of neutralizing antibodies were as follows:AdHu5, 75.1%;AdHu26, 66.8%;AdC68, 32.1%.CONCLUSION:The prevalence rates and titers of neutralizing antibodies against AdC68 were the lowest among the 3 adenoviruses.There-fore, AdC68 serves as more suitable biological therapy vectors for CHB and PLC than AdHu5 and AdHu26.

Objective To evaluate the curative efficacy of multimodality for severe pulmonary infection (SPI) following kidney transplantation (KT).Methods Fifty-seven cases of SPI following KT were treated with multimodality therapy in our hospital between Jan.2014 and Jan.2017.The outcome and data were analyzed and evaluated retrospectively.Results Of these 57 patients,45 cases were cured (41 cases were alive with functioning grafts,and 4 cases had grafts loss).The pulmonary lesions in 4 cases of pulmonary fungal infection were improved and oral anti-fungal drugs were continuously given after discharge.The symptoms in one case of tuberculosis were obviously improved and anti-tuberculosis treatment was given continuously after discharge.There were 5 deaths,including 2 deaths due to functioning grafts loss.Two cases abandoned treatment during therapy because of financial problem.Pathogens could be detected in only 29 cases.Conclusion SPI after KT is an acute important complication with rapid progression.Early and prompt treatment with combined antibiotics,antifungal drugs as well as antivirus is essential.The keys to successful rescue for SPI should also include immunosuppressant reduction,intravenous immunoglobulin and nutrition support.The combined therapy is successful and could reduce mortality of SPI obviously.

In recent years,the infection of nontuberculous mycobacterium(NTM)has been increasing rapidly,which captivates widespread attention.The infection rate of NTM in kidney transplant recipients is more significantly elevated due to the impact of immunosuppressive drugs and other factors.However,due to the lack of sufficient research evidence,relevant guidelines for the diagnosis and treatment of NTM after kidney transplantation are still lacking.To further standardize the diagnosis and treatment of NTM disease in kidney transplant recipients,and deepen medical practitioners'understanding and diagnosis and treatment of NTM disease in organ transplantation in China,Branch of Organ Transplantation of Chinese Medical Association organized relevant experts to formulate this guideline by referring to the latest edition of"An official ATS/IDSA statement:diagnosis,treatment,and prevention of nontuberculous mycobacterial diseases","Expert Consensus on the Diagnosis and Treatment of Nontuberculous Mycobacterial Disease",and"Technical Specification for Clinical Diagnosis and Treatment of Nontuberculous Mycobacteria in Organ Transplant Recipients(2019 Edition)",and considering the characteristics of kidney transplant recipients.

Chronic myeloid leukemia (CML) appears an ideal and exciting immunological target. Novel and rational immunotherapy may therefore play an important adjuvant role in the treatment of CML patients. Peptides derived from the BCR-ABL fusion region have been shown to be immunogenic and are able to stimulate the production of BCR-ABL-specific T cell lines and clones. In this study, A 280 bp multiple epitope region of BCR-ABL fusion antigen was designed and synthesized. This region contains three BCR-ABL antigen epitopes which can bind to HLA-A2, HLA-A3 and HLA-DR11 molecules, respectively, and epitopes of cholera toxin B (CTB) and tetanus toxoid (TT) which are able to elicit vigorous T cell responses. The fusion antigen gene has highly been expressed in E. coli and the purified fusion protein reserved satisfied activity and antigenicity. The results of this investigation provided a basis for further research on the developing specific T cell immunotherapy of CML.

Objective To evalute the role of phosphatidylinositol 3?kinase(PI3K)∕serine?threo?nine kinase(Akt)∕endothelial nitric oxide synthase(eNOS)signaling pathway in sevoflurane postcondi?tioning?induced attenuation of brain injury in a rat model of hemorrhagic shock and resuscitation(HSR). Methods Seventy?two pathogen?free healthy adult male Sprague?Dawleg rats, weighing 300-350 g, were divided into 4 groups(n=18 each)using a random number table: sham operation group(group S), group HSR, sevoflurane postconditioning group(group SP)and sevoflurane postconditioning plus PI3K∕Akt signaling pathway specific inhibitor wortmannin group(group SP+WT). Hemorrhagic shock was in?duced by withdrawing blood(40% of the total blood volume)from the right common carotid artery over an interval of 30 min, and 1 h later the animals were resuscitated with infusion of the shed blood via the left jugular vein over 30 min. In group SP+WT, wortmannin 0.6 mg∕kg was administrated via the jugular vein at 30 min before establishment of the model. In SP and SP+WT groups, 2.4% sevoflurane was inhaled for 30 min starting from the onset of infusion of the shed blood. At 10 min before withdrawing blood(T0), im?mediately after the end of withdrawing blood(T1), at 30 min and 1 h after the end of withdrawing blood (T2,3)and immediately after infusion of the shed blood(T4), blood samples from the common carotid ar?tery were collected for blood gas analysis, the blood lactate concentration was recorded, and mean arterial pressure was simultaneously recorded. At 24 h after infusion of the shed blood, 6 rats were randomly select?ed from each group and sacrificed, and their brains were immediately removed for determination of cerebral infarct volume(by TTC staining), expression of hippocampal caspase?3(by immuno?histochemistry), and expression of Akt, phosphorylated Akt(p?Akt)and eNOS(by Western blot). The ratio of p?Akt∕Akt was calculated. Results Compared with group S, the mean arterial pressure was significantly decreased and the blood lactate concentration was increased at T1?3, the cerebral infarct volume was increased, and the expression of caspase?3 was up?regulated in the other three groups, and the ratio of p?Akt∕Akt was sig?nificantly increased, and eNOS expression was up?regulated in group SP(P<0.05). Compared with group HSR, the cerebral infarct volume was significantly decreased, the expression of caspase?3 was down?regula?ted, the ratio of p?Akt∕Akt was increased, and eNOS expression was up?regulated in group SP(P<0.05). Compared with group SP, the cerebral infarct volume was significantly increased, the expression of caspase?3 was up?regulated, the ratio of p?Akt∕Akt was decreased, and eNOS expression was down?regula?ted in group SP+WT(P<0.05). Conclusion PI3K∕Akt∕eNOS signaling pathway activation mediates sevoflurane postconditioning?induced attenuation of brain injury in a rat model of HSR.

Objective To evaluate the role of mitochondrial permeability transition pore (mPTP)in reduction of brain injury by sevoflurane postconditioning in a rat model of hemorrhagic shock and resuscitation (HSR).Methods Ninety pathogen-free healthy adult male Sprague-Dawley rats,weighing 300-350 g,were divided into 5 groups (n =18 each) using a random number table:sham operation group (group S),group HSR,sevoflurane postconditioning group (group SP),sevoflurane postconditioning plus atractyloside (ATR,a specific mPTP opener) group (group SP + ATR) and ATR group.Hemorrhagic shock was produced by withdrawing 40％ of the total blood volume from the right carotid artery over an interval of 30 min,and 1 h later the animals were resuscitated by infusion of the shed blood via the left jugular vein over 30 min.SP and SP+ATR groups were exposed to 2.4％ sevoflurane for 30 min starting from the onset of reinfusion.In ATR and SP+ATR groups,ATR 5 mg/kg was intravenously injected at 10 min before reinfusion.Six rats in each group were randomly sacrificed at 24 h after the end of autologous blood reinfusion,and the hippocampus was harvested for determination of the expression of Bcl-2 and Bax in hippocampal tissues (by Western blot) and degree of mPTP opening.At 72 h after the end of autologous blood reinfusion,the rest 6 rats in each group were selected and underwent Morris water maze test,and the cognitive function was evaluated.Results Compared with group S,the escape latency was significantly prolonged,the number of crossing the original platform and locomotor distance in the target quadrant were decreased,the expression of Bcl-2 was down-regulated,the expression of Bax was up-regulated,and the degree of mPTP opening was increased in group HSR (P＜0.05).Compared with group HSR,the escape latency was significantly shortened,the number of crossing the original platform and locomotor distance in the target quadrant were increased,the expression of Bcl-2 was up-regulated,the expression of Bax was down-regulated,and the degree of mPTP opening was decreased in group SP (P＜0.05),and no significant change was found in each parameter in ATR and SP+ATR groups (P＞0.05).Compared with group SP,the escape latency was significantly prolonged,the number of crossing the original platform and locomotor distance in the target quadrant were decreased,the expression of Bcl-2 was down-regulated,the expression of Bax was up-regulated,and the degree of mPTP opening was increased in group SP+ATR (P＜0.05).Conclusion The mechanism by which sevoflurane postconditioning ameliorates brain injury may be related to inhibiting mPTP opening in a rat model of HSR.