Promotion of immune response by soluble Tim-3 in vitro and therapeutic potential / 军事医学

Objective To evaluate the role of recombinant human soluble Tim-3 (hTim-3-Fc) in regulating immune response.Methods Soluble hTim-3 was incubated with human macrophage cell line U 937, human T cell line Jurkat and normal human PBMC before cytokines secreted by or expressed in different immune cells were analyzed using ELISA , RT-PCR and Western-blotting, respectively.Results Soluble hTim-3 significantly promoted the activation of different immune cells.Our data showed that IL-8 secretion by U937 cells, IL-2 secretion by Jurkat cells , IL-2 and IFN-γsecretion by human PBMCs were all significantly increased .In addition , soluble hTim-3 significantly increased the IFN-α2 and IFN-β1 mRNA expression in U937, Jurkat and PBMCs and increased the phosphorylation of stat-1 in Jurkat and U937 cells.Conclusion Recombinant soluble hTim-3 can significantly promote the activation of immune cells in vitro, which shows its therapeutic potential .

Analysis on common complications during anesthesia recovery period after deep brain stimulation in patients with Parkinson disease / 中国医师进修杂志

Objective To investigate the common complications during anesthesia recovery period after deep brain stimulation in patients with Parkinson disease. Methods The complications during anesthesia recovery period after deep brain stimulation in 869 patients with Parkinson disease were retrospectively analyzed. Results The median recovery time during anesthesia recovery period was 15 (0 - 30) min. The complications of 869 patients with Parkinson disease were hypertension in 133 cases (15.3%), arrhythmia in 99 cases (11.4%), agitation in recovery period in 76 cases (8.7%), respiratory obstruction in 24 cases (2.8%), postoperative nausea and vomiting (PONV) in 18 cases (2.1%), hypoxemia in 17 cases (2.0%), pain in 10 cases (1.2%), delayed emergence in 10 cases (1.2%), shivering in 3 cases (0.3%), and hypotension in 1 case (0.1%). The incidence of 1 complication was 26.8%(233/869), the incidence of ≥ 2 complications was 9.9% (86/869), and the total incidence of complications was 36. 7% (319/869). In the 869 patients, the modified Aldrete score ≥ 9 scores when patients were removed away from the anesthesia recovery room was in 849 cases (97.7%), and≤8 scores was in 20 cases (2.3%). Conclusions The common complications during anesthesia recovery period after deep brain stimulation in patients with Parkinson disease are special. To maintain a stable circulation, preventing respiratory complications and aspiration are important to reduce the incidence and improve the prognosis.

Effect of estrogen on IL-10 production in mouse splenic B cells in vitro / 国际药学研究杂志

Objective To ascertain whether estrogen could induce B cell to produce interleukin(IL)-10. Methods C57BL/6 splenic B cells were purified by magnetic activated cell sorting(MACS)method,followed by estrogen treatment for 3 days. The secretion of IL-10 from cultured cell supernatant was tested by ELISA technique. The abundance of mRNA for IL-10、PD-L1 and RBM47 in B cells with estrogen treatment was tested by real-time PCR method. The intracellular IL-10 expression and the surface PD-L1 expression of treated B cells were tested by fluorescence activated cell sorting(FACS)method. And the expression of RBM47 in B cells by estrogen treatment was tested using Western blotting method. Results Estrogen could induce B cell to produce IL-10 in a dose-dependent manner. Estrogen treatment could increase the percentage of IL-10+B cells,the abundance of mRNA for IL-10,PD-L1 and RBM47 in B cells,as well as the expression of PD-L1 on B cell surface. Furthermore,our experimental result indicated the upreg?ulation of RBM47 expression in B cells by estrogen treatment. Conclusion Estrogen treatment in vitro can induce the upregulation of IL-10+regulatory B cells(Breg). Upregulation of RBM47 in the treated B cells might participate in this process by stabilizing IL-10 mRNA.

Roles of Tim-3 in paraquat poisoning induced acute inflammatory response / 军事医学

Objective To examine whether Tim-3 plays a protective role in paraquat poisoning induced excessive immune response and tissue damage based on the critical roles of Tim-3 controlling inflammatory response.Methods A paraquat poisoning model was established in wild type and in Tim-3 transgenic C57BL/6 mice by intraperitoneal injection of paraquat (40 mg/kg) .In addition, C57BL/6 mice with paraquat poisoning were injected with Tim-3 soluble protein( sTim-3) or control protein to see the effect of Tim-3 blocking on the progression of paraquat poisoning.Samples were collected at 6 and 24 h after paraquat injection respectively and were examined for tissue damage, cytokine expression and paraquat metabolism.Results After paraquat poisoning, there was significantly attenuated tissue damage in the lungs and kidneys and decreased TNF-α,IL-6 and IL-1 beta expression in the PBMCs or in the serum from Tim-3 transgenic mice compared to wild type mice.The serum concentration of paraquat in Tim-3 transgenic mice was also significantly decreased.However, in sTim-3 treated paraquat poisoning mice, there was significantly increased cytokine expression and tissue damage compared to control protein treated mice.The in vitro data showed that Tim-3 signaling negatively regulated macrophages mediated inflammatory response.Conclusion Tim-3 plays a critical role in maintaining the homeostasis after paraquat poisoning. Further investigation on the regulatory roles of Tim-3 in inflammation will shed new light on the pathogenesis of paraquat poisoning and provide new therapeutic strategies.

Postoperative Complications and Related Risk Factors in Neurosurgery Patients during Anesthesia Recovery / 中国康复理论与实践

Objective To investigate the characteristics of postoperative complications and related risk factors in neurosurgery patients. Methods A retrospective study was carried out in neurosurgery patients during anesthesia recovery period from March, 2009 to November, 2013. The recorded complications included respiratory and circulatory system complications, pain, shivering, nausea and vomiting, agitation and delaying recovery. Multivariate logistic regression analysis was performed to screen the risk factors for these complications. Results Da-ta of 13,495 patients were available for analysis. The general incidence was 48.8%. Post-operative complications included post-operative nausea and vomiting (PONV) (14.5%), agitation (13.5%), hypertension (13.4%), arrhythmia (9.3%), shivering (8.9%), pain (5.9%), hypox-emia (2.5%), delayed recovery (1.9%), airway obstruction (1.7%) and hypotension (0.3%). Regression analysis showed that the risk factors for hypoxemia included male,<59 years old and infratentorial tumor (P<0.05);the risk factors for PONV included male, supratentorial tu-mor, infratentorial tumor, cerebrovascular disease, inhalation anesthesia, the use of muscle relaxants antagonism and tramadol (P<0.05);and the risk factors for postoperative restlessness included male, infratentorial tumor and cerebrovascular disease, inhalation anesthesia, the use of muscle relaxants antagonism and no use of patient-controlled analgesia (PCA) pump (P<0.05). Conclusion Patients with different neuro-logical diseases showed different post-operative complications and exhibited different risk factors for these complications. Anesthesiologists should closely monitor patients receiving various neurosurgery and provide timely treatment.

Death patterns of patients in Department of Respiratory Care Unit of General Hospi-tal of PLA / 军事医学

Objective To analyze retrospectively the death pattern, risk factors, and death time of 253 patients at the Respiratory Care Unit of General Hospital of PLA in order to improve care quality and reduce mortality.Methods The information of patients was extracted from the hospital information system ( HIS) , and then classified and calculated accord-ing to different time points.Results Between November and next March,the mortality rate was higher than in other months (P<0.05), accounting for 19.5%.Mortality of those admitted between 8∶01 and 9∶00 or between 23∶01 and 24∶00 was higher than at other times(P<0.05), accounting for 41.7%and 50.0%, respectively.There was statistically significant difference(P<0.01) in mortality between days of the week,with the highest on Saturday, accounting for 43.1%.Mortality on non-work days was higher than on workday(P<0.01), accounting for 38.3% and 13.2%, respectively.Mortality at off-hour was higher than at office time(8∶00-11∶30 and 14∶30-18∶00 on workday) (P<0.01), accounting for 31.3%and 5.2%, respectively.Logistic regression analysis showed that age, month of admission, and the hour of discharge were associated with the outcome.Conclusion The high mortality between November and next March may be related to the higher incidence of respiratory diseases in winter, air pollution and cold weather.High mortality is also significantly associ-ated with the care quality of the medical staff.

Biological activity and application study of a monoclonal antibody against human Tim-3 / 军事医学

Objective To develop a human Tim-3 specific monoclonal antibody and evaluate its biological activity and possible use in clinical diseases associated with dysregulated Tim-3 expression .Methods The BALB/c mice were immu-nized by conventional method, and positive clones were used to develop anti-human Tim-3 antibody, the binding and neutralization activities of which in vitro and in vivo were investigated.Results ①A monoclonal antibody (clone L3D) which could specifically bind to human Tim-3 protein in ELISA assay was obtained and the subtype of the monoclonal antibody was IgG2a .②Flow cytometry indicated that the monoclonal antibody could bind to Tim-3 expressed in human U937 cells.This antibody also showed a cross activity to mice′Tim-3.③The monoclonal antibody inhibited the apoptosis of THP1 cells induced by Gal-9, the ligand of Tim-3.④Injection of Tim-3 antibody exacerbated sepsis in mice as marked by the decreased survival rate and increased expression of pro-inflammatory cytokines .Conclusion An anti-human Tim-3 monoclonal antibody is successfully obtained.The excellent binding and neutralization activities of this antibody enable it to be widely used in clinical diseases associated with deregulated Tim-3 expression .

Ligament reconstruction tendon interposition arthroplasty for first carpometacarpal joint osteoarthritis / 中华骨科杂志

Objective To explore the clinical outcomes of Ligament reconstruction tendon interposition (LRTI) arthro-plasty for first carpometacarpal joint osteoarthritis. Methods From January 2008 to January 2011, 19 patients (21 thumbs) had surgery for thumb carpometacarpal arthritis using ligament reconstruction tendon interposition arthroplasty with flexor carpi radia-lis (FCR). There were 1 male and 18 were females with an average age of 60 years (range, 52-75 years);8 thumbs were on the left side and 13 thumbs on the right side. According to Eaton-Glickel classification, 1 thumb belonged to stageⅡ, 14 thumbs to stageⅢ, and 6 thumbs to stageⅣ. Pain level, grip strength, tip pinch strength, range of motion, and radiographic measurement were re-corded. According to the first metacarpal subsidence, the cases were classified in mild, moderate, and severe groups. Clinical out-comes of different group were evaluated and compared. Results All patients were followed up for 9-28 months with an average of 13.9 months. Comparision with the preoperative X-rays showed the first metacarpal had subsided 54.8% of the arthroplasty space after surgery. Grip strength improved from 18.6±10.1 kg to 20.5±11.9 kg, and tip pinch strength increased from 4.4±2.1 kg to 4.5 ± 1.9 kg after the surgery. Radial abduction increased from 55.7° ± 8.2° to 60.6° ± 7.2° and palmar abduction improved from 56.7° ± 8.5° to 63.5° ± 8.2° after the procedure. Patient pain levels (visual analogue scale, VAS) were significantly reduced, from 6.6 ± 1.4 to 0.5 ± 0.7. There was no difference of grip strength, tip pinch strength, thumbs range of motion, and VAS after LRTI in mild, moderate and severe groups. Conclusion LRTI resulted in excellent relief of pain and increase in range of motion. Howev-er, LRTI cannot sustain the arthroplasty space. Compared with the preoperative X-ray, the first metacarpal subsided more than 50%. Subsidence of the first metacarpal doesn't affect the pain relief, range of motion and strength improvement.

Analysis of postoperative residual paralysis and respiratory function of patients undergoing neurosurgical anesthesia / 中国医师进修杂志

ObjectiveTo investigate the incidence of postoperative residual paralysis and respiratory function of patients undergoing neurosurgical anesthesia.Methods Three hundred andtwenty-eight patients undergoing neurosurgical anesthesia (ASA Ⅰ - Ⅱ ) were divided into vecuronium group (135 cases) and rocuronium group (193 cases) by random digits table.Anesthesia was maintained with propofol in combination with sevoflurane and intermittented intravenous infusion boluses of muscle relaxatant.Extubation of the intratracheal tube in operation room was performed under clinical criteria.The neuromuscular function were detected by acceleromyography with supramaximal train-of- four(TOF) stimulation of the ulnar nerve,and pulmonary function were measured in postanesthesia care unit(PACU).The TOF ratio and pulmonary function were compared between two groups.ResultsThe incidence of postoperative residual paralysis was 31.9％(43/135) in vecuronium group,and 14.5％(28/193) in rocuronium group.Hypoxemia and hypercapnia occurred in vecuronium group with TOF ＜ 0.9 were higher than in those with TOF ≥0.9[7.0％(3/43) vs.4.3％(4/92) and 44.2％(19/43) vs.18.5％(17/92),P ＜ 0.01 ],while in rocuronium group with TOF ＜ 0.9 were higher than in those with TOF ≥ 0.9 [ 3.6％( 1/28 ) vs.1.2％ (2/165) and 39.3％( 11/28 )vs.17.0％ (28/165),P ＜ 0.01 ].ConclusionsThere is a high incidence of postoperative residual paralysis according to the clinical criteria of recovery of neuromuscular function of patients undergoing neurosurgical anesthesia,which would impair respiratory function.

Comparison of preventive effects of tramadol and ketamine on remifentanil-induced postoperative hyperalgesia after neurosurgical anesthesia / 中国医师进修杂志

ObjectiveTo compare the preventive effects oftramadol and ketamine on the patients with postoperative hyperalgesia after remifentanil-induced neurosurgical anesthesia.MethodsOne hundred and fifty patients undergoing craniotomy were assigned to tramadol group,ketamine group and normal saline group with 50 cases each by random digits table.Anesthesia was maintained with infusion of remifentanil [0.1-0.2μ g/( kg· min ) ],propofol and sevoflurane.Tramadol ( 1.5 mg/kg),ketamine (0.5 mg/kg) or normal saline was given before skin closing.The emergence time,trachea extubation time,patients required analgesia and tramadol consumption,reverse effect were recorded.The visual analog scale (VAS),Ramsay scores at 15,30,60,120 minutes after emergence were performed.ResultsPatients required analgesia and tramadol consumption in tramadol group and ketamine group were significantly lower than those in normal saline group (P ＜0.01 or ＜0.05 ).The occurrence of shiver in tramadol group was lower than that in normal saline group (P ＜0.05).VAS scores in tramadol group at 15,30 minutes and in ketamine group at 30 minutes after emergence were significantly lower than those in normal saline group (P＜ 0.05).Ramsay score in ketamine group at 15 minutes after emergence was higher than that in tramadol group and normal saline group [ (2.9 ±0.6) scores vs.(2.3 ±0.7) scores and (2.3 ±0.9) scores](P＜0.01).ConclusionTramadol has a goodpreventive effect for postoperative hyperalgesia induced by remifentanil in neurosurgery compared with ketamine.

Incidence and Prevention of Postoperative Hypoxemia in Transsphenoidal Pituitary Tumor Resection / 中国康复理论与实践

@#ObjectiveTo observe the incidence and mechanism of postoperative hypoxemia in the patients after transsphenoidal pituitary tumor resection.Methods579 postoperative patients in postanesthesia care unit (PACU) in March, 2009～June, 2010 were investigated retrospectively using PACU databank. The incidence and mechanism of hypoxemia were analyzed.ResultsThe incidence of hypoxemia was 5.2%. Upper airway obstruction, change of breath mode, residual effects of anesthetics, bronchiospasm, aspiration, acute pulmonary edema and lung disease were the main reasons of postoperative hypoxemia after transsphenoidal pituitary tumor resection.ConclusionCareful monitor and treatment are the key to prevent hypoxemia after transsphenoidal pituitary tumor resection.

Postoperative Complications after Intracranial Neurosurgery in the Postanesthesia Care Unit / 中国康复理论与实践

@#ObjectiveTo investigate the complications after intracranial neurosurgery in the postanesthesia care unit (PACU). Methods2166 patients enrolled into PACU after intracranial neurosurgery were reviewed. ResultsAmong the complications, the most frequent ones were hypertension (14.0%), arrhythmia (13.9%), pain (13.7%), shivering (10.5%), nausea and vomiting (9.3%), and delirium (8.6%). The least ones were hypoxemia (3.4%), respiratory obstruction (1.9%), delayed recovery (1.5%), and hypotention (0.5%). Abnormal temperature and residual block occurred at 7.2% and 22.8% in the patients who were monitored. All complications were treated immediately, and all the patients transferred to neurosurgical ward with modified Aldrete score over 9～10. ConclusionThe patient in PACU need well management to insure safe and smooth recovery from anesthesia after intracranial surgery.

Effectiveness analysis on hysteroscopy combined with Chinese traditional medicine treating obstructive fallopian tube infertility / 中国医师进修杂志

Objective To discuss the effect of treating tubal obstruction infertility through hydrotubation by hysteroscopy combined with Chinese traditional medicine. Methods Chose 308 patients with tubal obstruction infertility from October 2001 to December 2008 and used the hysteroseopy under which making tubal cannulation, then injected methylene blue and clear liquid to record cases of tubal patency.After the operation the patients would be given Chinese traditional medicine and followed up the case of pregnancy. Result The recanalization rate in 308 patients was 90.9% (280/308), and the rate ofre-pregnancy was 55.5%(161/290). Conclusion The effect of treatment through hysteroscopy to make tubal cannulation and have patients take medicine after operation is very satisfactory and it is worth promoting.

Construction of Expression Vector for Human CD40-Ig Fusion Protein and Its Expression in COS-7 Cells / 中国实验血液学杂志

CD40/CD40L, besides B7/CD28, is an alternative important costimulation signal transduction pathway. It plays a pivotal role in T cell activation. Moreover, it may play a critical role at many levels of sensitization and effector phases of allograft rejection. In order to get the fusion protein of human CD40 extracelluar region and IgG 1 Fc fragment, and investigate the potential role of blocking CD40/CD40L costimulation pathway in immunotherapy, total RNA was extracted from human lymphoma cell line Daudi, and CD40 gene extracelluar region was amplified by RT-PCR. The PCR products were inserted into pGEM T Easy vector, and the cloning vector pGE40 was obtained. The DNA sequence was analyzed by automatic DNA sequencer. After sequencing, the transient expressing vector was constructed by inserting correct fragment into pIG vector, which contains the genomic human IgG1 Fc (hinge, CH2 and CH3) gene. Hence the recombinant fusion expression vector was constructed successfully, and named after pIG/40 Ig. Then, COS-7 cells were transfected through DEAE-Dextran/chloroquine method. The CD40-Ig fusion protein expressed in COS-7 cell culture supernatant was identified by sandwich ELISA and Western blot. Result showed that the CD40-Ig fusion protein can be detected by sandwich ELISA in the cell culture supernatant. Western blot analysis also showed that it could react with McAbs of mouse anti-human CD40 G28-5 and mouse anti-human Ig gamma chain. There is only one obvious band at the position of relative molecular weight 50 kD, and it is equivalent to the expected value. Above all, the recombinant fusion expression vector pIG/40 Ig was constructed, and CD40-Ig fusion protein gene was expressed in COS-7 cells successfully. It could be laid a foundation to investigate the potential role of CD40/CD40L pathway as the target of GVHD prevention and therapy.

The usage of abdominal double layer subcutaneous flap in repairing late complicated hand injury / 中华骨科杂志

Objective To introduce the method, indications and the usage of abdominal double layer subcutaneous flap for the repair of late complicated hand injury. Methods A new type of flap was designed to repair the complicated hand injury. Sixteen cases were treated with this method since 1998. Results The duration of the follow up ranged from 3 to 24 months with an average of 13 months. All of the flaps survived well. After physiotherapy, the overall excellent and good rate of joint movement was 80%(TAM standard). No further surgery was required in all of the cases. Conclusion The abdominal double layer subcutaneous flap was especially indicated in those cases with bone nonunion, tendon and nerve defect and tendonous adhesions. This kind of flap not only provide a good skin covering, but also offer a soft bed for the tendons, nerves, and bones.

Antitumor Responses Induced by Recombinant Vaccinia Viruses Expressingp53 Antigenic Peptide and B7 / 中国肿瘤生物治疗杂志

Objective: To explore antitumor responses induced by recombinant vaccinia viruses expressing a p53 antigenic peptide (rVV-p53M) and enhanced effect of recombinant vaccinia viruses expressing costimulatory molecule B7 (rVVB7). Methods: A 135 Cys to Tyr point mutation p53-transduced P815 mastocytoma (P815-mp53) was used as an experimental tumor and the p53 protein as the model of tumor associated antigen． rVV-p53M and rVV-B7 were used as vaccine to test their induction of CTL and antitumor immunity. Results: Immunization of BABL/c mice with rVV-p53M could elicit specific CTLs, which could specifically lyse P815-mp53 cells. Immunization of mice with rVV-p53M could survive a part of mice challenged with 1×106 P815-mp53. Treatment with rVV-p53M could significantly prolong the survival oftumor-bearing mice. Admixture at 1:1 ratio of rVV-p53 M and rVV-B7 could enhance antitumor responses induced by rVV-p53M. Conclusion: Immunization with recombinant vaceinia virus expressing antigenic peptide is a useful alternative to peptide-based vaccine. Costimulatory molecule B7 can enhance antigenic peptide to induce antitumor responses.

Antitumor Responses Induced by Recombinant Vaccinia Viruses Expressing p53 Antigenic Peptide and B7 / 中国肿瘤生物治疗杂志

Objective: To explore antitumor responses induced by recombinant vaccinia viruses expressing a p53 antigenic peptide (rVV p53 M) and enhanced effect of recombinant vaccinia viruses expressing costimulatory molecule B7 (rVV B7). Methods: A 135 Cys to Tyr point mutation p53 transduced P815 mastocytoma (P815 mp53) was used as an experimental tumor and the p53 protein as the model of tumor associated antigen. rVV p53 M and rVV B7 were used as vaccine to test their induction of CTL and antitumor immunity. Results: Immunization of BABL/c mice with rVV p53 M could elicit specific CTLs, which could specifically lyse P815 mp53 cells. Immunization of mice with rVV p53 M could survive a part of mice challenged with 1?10 6 P815 mp53. Treatment with rVV p53 M could significantly prolong the survival of tumor bearing mice. Admixture at 1∶1 ratio of rVV p53 M and rVV B7 could enhance antitumor responses induced by rVV p53 M. Conclusion: Immunization with recombinant vaccinia virus expressing antigenic peptide is a useful alternative to peptide based vaccine. Costimulatory molecule B7 can enhance antigenic peptide to induce antitumor responses.

The Therapeutic Effects of Intratumoral Injection with MHC Class Ⅱ Gene on Tumor-Bearing Mice / 中国肿瘤生物治疗杂志

Many gene therapy protocols can induce antitumor immunity, however, the ex vivo approach restricts their uses. This sutydy intended to induce antitumor immunity by direct transfer of MHC class Ⅱ gene in vivo. Methods: MHC class Ⅱ gene cDNA was introduced directly into two tumors: P815, (a murine weak immunogenic mas-tocytoma) and B16 (a murine nonirnmunogenic melanoma) to observe the survival rate of the mice. Results: Tumori-genicity of P815 was reduced when MHC class Ⅱ gene was introduced directly into tumors in vivo. Further more, most vaccinated mice could survive after second challenge of P815. Co-injection of MHC class Ⅱ and B7 genes in the B16 also resulted in the tumor grow slowly, while the injection of MHC class Ⅱ gene was not enough to induce effective antitumor responses. Conclusion: The results showed the potential applications of direct transfer of MHC class Ⅱ gene in the treatment of tumor.