Effect of electroacupuncture at the acupoints for Tiaozang Xingshen on cerebral metabolism in the patients with type 2 diabetes mellitus-associated cognitive dysfunction / 中国针灸

OBJECTIVES@#To investigate the cerebral metabolism in the patients with type 2 diabetes mellitus-associated cognitive dysfunction (T2DACD) and explore the mechanism of electroacupuncture (EA) at the acupoints for Tiaozang Xingshen (adjusting zangfu function and rescuing the spirit) in treatment of T2DACD, using magnetic resonance spectroscopy.@*METHODS@#Fifteen patients with T2DACD (observation group) and 22 healthy subjects (control group) were enrolled. In the observation group, the patients were treated with EA for Tiaozang Xingshen at Baihui (GV 20) and Shenting (GV 24), and bilateral Feishu (BL 13), Pishu (BL 20), Shenshu (BL 23), Zusanli (ST 36), Sanyinjiao (SP 6), Hegu (LI 4) and Taichong (LR 3). EA was operated with disperse-dense wave, 2 Hz/100 Hz in frequency and 0.1 mA to 1.0 mA in current intensity; 30 min each time, once daily. One course of EA consisted of 5 treatments, at the interval of 2 days and the intervention lasted 8 courses. Before treatment in the control group, before and after treatment in the observation group, the score of Montreal cognitive assessment scale (MoCA), the score of clinical dementia rating (CDR), Flanker paradigm, Stroop paradigm, Nback paradigm, the score of self-rating anxiety scale (SAS), the score of self-rating depression scale (SDS), and the score of Hamilton depression rating scale (HAMD) were evaluated separately; the glycolipid metabolic indexes (fasting plasma glucose [FPG], glycosylated hemoglobin type A1c [HbA1c], total cholesterol [TC], triacylglycerol [TG], high-density lipoprotein cholesterol [HDL-C] and low-density lipoprotein cholesterol [LDL-C]) were determined；with the magnetic resonance spectroscopy technique adopted, the metabolites in the basal ganglia area were detected. The correlation analysis was performed for the metabolite values with MoCA score, CDR score , Flanker paradigm, Stroop paradigm, and Nback paradigm.@*RESULTS@#Before treatment, compared with the control group, in the observation group, MoCA score was lower (P<0.001), CDR score and the levels of FPG and HbA1c were higher (P<0.001); the reaction times of Flanker non-conflict, Flanker conflict, Stroop neutrality, Stroop congruence, Stroop conflict, and 1-back were prolonged (P<0.05, P<0.001), and the accuracy of Flanker conflict, Stroop conflict, and 1-back decreased (P<0.05, P<0.01); the ratio of N-acetyl aspartate (NAA) to creatine (Cr) in the left basal ganglia area was dropped (P<0.001), and that of myo-inositol (MI) to Cr in the right side increased (P<0.05). In the observation group after treatment, compared with the levels before treatment, MoCA score was higher (P<0.001), the scores of CDR, SAS and HAMD were reduced (P<0.01, P<0.05), the reaction times of Flanker conflict and Stroop conflict shortened (P<0.001, P<0.05), and the accuracy of Flanker conflict and 1-back increased (P<0.001, P<0.05); the ratio of NAA to Cr in the left basal ganglia area and that of the gamma-aminobutyric acid (GABA) to Cr in the right increased (P<0.05), that of MI to Cr in the right decreased (P<0.05). Before treatment, in the observation group, the ratio of MI to Cr in the right basal ganglia area was positively correlated with the reaction time of Stroop congruence (r=0.671, P=0.012) and this ratio was positively correlated with the reaction time of Stroop conflict (r=0.576, P=0.039).@*CONCLUSIONS@#Electroacupuncture for "adjusting zangfu function and rescuing the mind" improves the cognitive function of T2DACD patients, which may be related to the regulation of NAA, MI and GABA levels in the basal ganglia.

Research progress in pH-responsive antibacterial nanomaterials for inhibiting oral plaque biofilms / 中华口腔医学杂志

pH-responsive antibacterial nanomaterials are a new type of nanomaterials that can selectively undergo structural changes and trigger drug release according to significant pH differences produced by the body under physiological and pathological conditions. The formation of acidic microenvironment in plaque is the key to the pathogenicity of oral plaque biofilm, which also creates conditions for the oral application of pH-responsive antibacterial nanomaterials. pH-responsive antibacterial nanomaterials can respond to changes in pH of plaque microenvironment and accurately control the release of antibacterial drugs, providing a new direction for improving drug efficacy and targeted antibacterial. The present article reviews the classification, mechanism and application of pH-responsive antibacterial nanomaterials in oral plaque inhibition.

Significance of early lymphocyte-platelets ratio on the prognosis of patients with extensive burns / 中华烧伤杂志

Objective: To analyze the changing trend and characteristics of lymphocyte-platelets ratio (LPR) of early stage in patients with extensive burns, and to explore the prognostic significance of LPR. Methods: A retrospective case series study was conducted. From January 2008 to December 2018, 244 patients with extensive burns were admitted to the First Affiliated Hospital of Naval Medical University, including 181 males and 63 females, aged (44±16) years. The total burned area of patients was 60.0% (42.0%, 85.0%) total body surface area. Platelet and lymphocyte test results of patients were collected on the 1st, 2nd and 3rd day after admission, and LPR of patients was calculated to analyze the changing trend of the three days after admission. Univariate and multivariate logistic regression analysis were conducted to investigate the risk factors or independent risk factors for death of patients, including age, sex, total burn area, area of full-thickness burns and above, inhalation injury, and LPR. According to the 1st day's LPR after admission of patients, the receiver operating characteristic (ROC) curve predicting death of patients was drawn to find the optimal value of LPR. Patients were divided into high LPR group (n=136) and low LPR group (n=108) based on the optimal value of LPR, and the clinical data of total burn area, area of full-thickness burns and above, inhalation injury, tracheotomy, offline time of patients within 28 days, and mortality in the 2 groups were compared. The surviving curve of patients was drawn by Kaplan-Meier method to predict the difference of the 90-day survival rate between the two groups of patients. Data were statistically analyzed with Student's t test, Mann-Whitney U test, and chi-square test. Results: Within 3 days of admission, the LPR of patients showed a time-dependent upward trend. LPR of patients on the 2nd and 3rd day after admission was 8.6 (5.3, 14.4) and 8.6 (4.9, 13.7), respectively, which were significantly higher than the 1st day's 6.3 (4.2, 9.8), with Z values of -4.25 and -3.43, respectively, P<0.01. Univariate logistic regression analysis showed that age, total burn area, area of full-thickness burns and above, inhalation injury, and LPR were all risk factors for death of patients (with odds ratios of 1.03, 1.73, 1.31, 4.74, and 3.11, respectively, 95% confidence intervals of 1.01-1.06, 1.40-2.13, 1.21-1.42, 1.62-13.86, and 1.41-6.88, respectively, P<0.01). Multivariate logistic regression analysis showed that age, area of full-thickness burns and above, and LPR were independent risk factors for death of patients (with odds ratios of 1.06, 1.36, and 2.85, respectively, 95% confidence intervals of 1.03-1.09, 1.19-1.55, 1.02-7.97, P<0.05 or P<0.01). The area under ROC curve of the 1st day's LPR, predicting death of patients, was 0.61 (with 95% confidence interval of 0.51-0.71, P<0.05), and the optimal predicted value was 5.8 with corresponding sensitivity of 77% and specificity of 52% respectively. The total burn area, area of full-thickness burns and above, rates of incidence of inhalation injury, tracheotomy, and mortality of patients in high LPR group were significantly higher than those in low LPR group (with Z values of -3.06 and -3.19, χ2 values of 5.42, 11.64, and 8.45, respectively, P<0.05 or P<0.01). The offline time of patients within 28 days in high LPR group was significantly shorter than that in low LPR group (Z=-2.98, P<0.01). Kaplan-Meier survival analysis showed that the 90-day survival rate of admission of patients in low LPR group was significantly higher than that of patients in high LPR group (χ2=8.24, P<0.01). Conclusions: The early LPR of patients with extensive burns showed a time-dependent upward trend. The LPR on the first day after admission that is closely correlated with total burn area, area of full-thickness and deeper burns, inhalation injury, tracheotomy, and mortality of patients, is an independent risk factor for the prognosis of patients with extensive burns. The first day's LPR after admission is significantly correlated with the 90-day survival rate of patients, which can be used as an evaluation index for the severity of extensive burns.

Gut-derived lipopolysaccharide promotes alcoholic hepatosteatosis and subsequent hepatocellular carcinoma by stimulating neutrophil extracellular traps through toll-like receptor 4

Background/Aims@#Binge drinking leads to many disorders, including alcoholic hepatosteatosis, which is characterized by intrahepatic neutrophil infiltration and increases the risk of hepatocellular carcinoma (HCC). Molecular mechanisms may involve the migration of bacterial metabolites from the gut to the liver and the activation of neutrophil extracellular traps (NETs). @*Methods@#Serum samples from both binge drinking and alcohol-avoiding patients were analyzed. Mouse models of chronic plus binge alcohol-induced hepatosteatosis and HCC models were used. @*Results@#A marker of NETs formation, lipopolysaccharide (LPS), was significantly higher in alcoholic hepatosteatosis and HCC patients and mice than in controls. Intrahepatic inflammation markers and HCC-related cytokines were decreased in mice with reduced NET formation due to neutrophil elastase (NE) deletion, and liver-related symptoms of alcohol were also alleviated in NE knockout mice. Removal of intestinal bacteria with antibiotics led to decreases in markers of NETs formation and inflammatory cytokines upon chronic alcohol consumption, and development of alcoholic hepatosteatosis and HCC was also attenuated. These functions were restored upon supplementation with the bacterial product LPS. When mice lacking toll-like receptor 4 (TLR4) received chronic alcohol feeding, intrahepatic markers of NETs formation decreased, and hepatosteatosis and HCC were alleviated. @*Conclusions@#Formation of NETs following LPS stimulation of TLR4 upon chronic alcohol use leads to increased alcoholic steatosis and subsequent HCC.

Clavien-Dindo classification and influencing factors analysis of complications after laparoscopic pancreaticoduodenectomy / 中华外科杂志

Objective@#To semi-quantify the postoperative complications occurred after laparoscopic pancreaticoduodenectomy(LPD) using Clavien-Dindo score, thereafter exploring its impact factors.@*Methods@#In this retrospective cohort study, the clinical data of 124 patients who had undergone LPD for periampullary tumor from June 2016 to June 2017 at Department of Biliary Pancreatic Surgery, Affiliated Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology were collected.Malignancy was confirmed based on postoperative pathological reports.Postoperative complications were semi-quantitated using Clavien-Dindo score.Multivariable logistic regression model was applied to explore the factors related to severe complications(Clavien-Dindo Ⅲb-Ⅴ).@*Results@#Of the 124 patients, there were 64 males(51.6%) and 60 females(48.4%), with age of 57 years(range, 23-82 years). In total, postoperative complications occurred in 30 patients(24.2%). Among the 30 patients, 4 patients suffered Clavien-Dindo grade Ⅰ, 18 patients(14.5%) suffered Clavien-Dindo grade Ⅱ, 6 patients(4.8%) suffered Clavien-Dindo grade Ⅲa, 1 patient(0.1%) suffered Clavien-Dindo grade Ⅳb, and 1 patient(0.1%) suffered Clavien-Dindo grade Ⅴ.Intraabdominal hemorrhage occurred in 8 patients, pancreatic fistula was found in 10 patients(7 patients had biochemical leakage and 3 of them had grade B pancreatic fistula), both biliary fistula and gastrointestinal fistula were found in 1 patient.Abdominal infection occurred in 10 patients, both liver failure and renal failure occurred in one patient.Moreover, arrhythmia was found in two patients, and mortality occurred in one patient.Five patients suffered multiple complications.Univariable analysis showed that postoperative complications were associated with body mass index, American Society of Anesthesiologists(ASA) score, intraoperative blood transfusion, and pancreatic texture(P<0.05). In multivariable logistic regression, ASA grade Ⅲ, intraoperative blood transfusion, and pancreatic softness were independently related to postoperative complications after LPD(P<0.05).@*Conclusions@#Clavien-Dindo score is feasible to be applied in management of patients with LPD.ASA score, texture of pancreas, and intraoperative blood transfusion were independently associated with postoperative complications.

Short-term outcome analysis of laparoscopic and open pancreaticoduodenectomy for pancreatic head cancer / 中华消化外科杂志

Objective To explore the short-term outcome of laparoscopic pancreaticoduodenectomy (LPD) and open pancreaticoduodenectomy (OPD) for pancreatic head cancer.Methods The retrospective cohort study was conducted.The clinicopathological data of 108 patients with pancreatic head cancer who were admitted to the Affiliated Tongji Hospital of Huazhong University of Science and Technology between July 2014 and July 2015 were collected.Among 108 patients,47 and 61 who respectively underwent LPD and OPD were allocated into LPD and OPD groups.Observation indicators:(1) intraoperative situations;(2) postoperative situations;(3) postoperative pathological situations;(4) follow-up and survival situations.Follow-up using outpatient examination and telephone interview was performed to detect chemotherapy and postoperative survival situations at 1 and 3 years postoperatively up to June 2018.Measurement data with normal distribution were represented as x±s and comparison between groups was analyzed using the t test.Comparison between groups of count data was analyzed using the chi-square test.Results (1) Intraoperative situations:operation time in the LPD and OPD groups was respectively (288±24)minutes and (265±29)minutes,with no statistically significant difference between groups (t=5.138,P＞0.05).Volume of intraoperative blood loss in the LPD and OPD groups was respectively (136±14)mL and (388±21)mL,with a statistically significant difference between groups (t=-7.297,P＜0.05).Cases with blood transfusion were respectively 3 and 7 iu the LPD and OPD groups,with no statistically significant difference between groups (x2 =0.325,P ＞ 0.05).(2) Postoperative situations:of 47 patients in the LPD group,16 with postoperative complications were improved by conservative treatment,including 7 with pancreatic fistula (5 with biochemical pancreatic fistula and 2 with grading B and C of pancreatic fistula);4 with delayed gastric emptying were cured by gastrointestinal decompression and gastric motility promoting treatment;2 with postoperative bleeding were improved by conservative treatment;2 with intraabdominal infection were improved by enhanced antibiotic therapy and transabdominal percutaneous drainagc;1 with biliary fistula was improved by transabdominal percutaneous drainage;there was no wound infection and perioperative death.Of 61 patients in the OPD group,28 with postoperative complications were improved by conservative treatment,including 12 with pancreatic fistula (9 with biochemical pancreatic fistula and 3 with grading B and C of pancreatic fistula);8 with delayed gastric emptying were cured by gastrointestinal decompression and gastric motility promoting treatment;3 with intra-abdominal infection were improved by enhanced antibiotic therapy and transabdominal percutaneous drainage;2 with postoperative bleeding were improved by conservative treatment;2 with wound infection were c ured by conservative treatment;1 with biliary fistula was improved by transabdominal percutaneous drainage;there was no perioperative death.There was no statistically significant difference in the cases with postoperative complications between groups (x2 =1.546,P＞ 0.05).Duration of hospital stay in the LPD and OPD groups was (13.6±2.1)days and (19.3 ±4.4)days,respectively,with a statistically significant difference (t =-4.354,P＜0.05).(3) Postoperative pathological situations:R0 resection rate was respectively 100.0％ (47/47) and 98.4％ (60/61) in the LPD and OPD groups,with no statistically significant difference (x2 =0,P＞0.05),and there was 1 patient with R1 resection in the OPD group.The total number of lymph node dissected in the LPD and OPD groups was respectively 19±4 and 13±4,with a statistically significant difference (t=-4.126,P＜0.05).The cases with high-and moderate-differentiated tumor and low-differentiated tumor (tumor differentiation),staging T1-T2 and T3-T4 (T stage),staging N0 and N1 (N stage),staging Ⅰ and Ⅱ-Ⅲ (TNM staging) and nerve or vascular invasion were respectively 35,12,28,19,20,27,16,31,21 in the LPD group and 50,11,36,25,36,25,14,47,32 in the OPD group,with no statistically significant difference (x2=0.891,0.003,2.882,1.628,0.643,P＞0.05).(4) Follow-up and survival situations:44 and 55 patients in the LPD and OPD group respectively underwent postoperative adjuvant therapy during the follow-up,with no statistically significant difference (x2=0,P＞0.05).The postoperative 1-year follow-up:47 patients in the LPD group were followed up,37 survived and 10 died;of 61 patients in the OPD group,3 lost to follow-up,and 58 were followed up (43 survived and 15 died);there was no statistically significant difference in survival between groups (x2=0.301,P＞0.05).The postoperative 3-year follow-up:of 47 patients in the LPD group,3 lost to follow-up,and 44 were followed up (21 survived and 23 died);of 61 patients in the OPD group,6 lost to follow-up,and 55 were followed up (23 survived and 32 died);there was no statistically significant difference in survival between groups (x2 =0.346,P＞0.05).Conclusion LPD is safe and feasible for pancreatic head cancer,with advantages of less bleeding,shorter duration of hospital stay and more total number of lymph node dissected,and its survival effect is equivalent to that of OPD.

The protective effect of rosiglitazone on the rats with high altitude pulmonary edema / 中华急诊医学杂志

Objective To investigate the protective effect of rosiglitazone on the rats with high altitude pulmonary edema.Methods Thirty-six SD rats were randomly (random number) divided into 6 groups (n =6 each):control group (Control),hypobaric hypoxia model group (HH),rosiglitazone groups (RSG) which were administered with 3 different doses [RSG-L:5 mg/ (kg · d),RSG-M:10 mg/ (kg·d),RSG-H:20 mg/ (kg· d)],dexamethasone group [Dex,4 mg/ (kg· d)].Rats were injected intraperitoneally with different doses of rosiglitazone (RSG),dexamethasone (Dex) or vehicle (Control and HH) for 3 days before placed in simulated altitude of 6 000 m hypobaric hypoxia animal chamber where the temperature and pressure were constant.After 72 h in the chamber,each rat was anesthetized.The water content of lung was determined with wet/dry weight ratio.Bronchoalveolar lavage fluid was measured by bradford method.The contents of GSH was measured by micro-ezymed labeled method.The contents of MDA was measured by TBA method.The enzymatic activities of SOD was measured by WST-1 method.The changes of the TNF-α,IL-6 and IL-10 in serum were determined by ELISA.Light microscope was used to observe the pathological changes of lung tissue.Results Compared with Control group,the wet/dry weight ratio of lung (5.08 ± 0.24) and total protein content of BALF (351.06 ± 44.55) μg/mL increased significantly (P ＜ 0.01) in HH group.There were red blood cells in the alveolar and interstitium,pink fluid exudation in the alveolar,the alveolar septum enhancement,and a large number of inflammatory cell infiltration;the SOD activity (10.65 ± 0.94) U/mgprot and the content of GSH (1.63 ±0.20) μmol/gprot in lung tissue were significantly decreased (P ＜ 0.01),the contents of MDA (2.1 5 ± 0.18) nmol/mgprot increased significantly (P ＜ 0.01),TNF-o (56.92 ± 2.87) pg/mL and IL-6 (217.80 ±48.01) pg/mL levels in serum were significantly increased (P ＜0.01),and IL-10 (76.85 ± 16.72) pg/mL level decreased (P ＜ 0.05).Compared with the HH group,the wet/dry ratio of lung and total protein content of BALF in different doses of rosiglitazone group significantly decreased (P ＜ 0.01),the pathological changes of the lung tissue was significantly improved,SOD activity and the content of GSH in lung tissue was significantly increased (P ＜ 0.01),the content of MDA decreased (P ＜ 0.01),The levels of TNF-α and IL-6 in serum were significantly decreased (P ＜ 0.01),while the IL-10 level was significantly increased (P ＜ 0.01).Conclusion Rosiglitazone could protect the high altitude pulmonary edema by alleviating the oxidative stress and inflammatory response.

Optimization of prokaryotic expression condition and purification of soluble GST-CRH protein / 天津医药

Objective To obtain the recombinant corticotropin releasing hormone (CRH) protein with soluble, high purity protein through optimizing prokaryotic expression condition and purifying glutathione thiol transferase (GST)-CRH protein. Methods To detect the expression of soluble CRH protein through grope of the host strain GST-CRH temperature of induction expression, the host strain concentration (OD600), IPTG concentration and induction time, the purification of GST-CRH was performed by GST-CRH agarose gel. Western Blot assay was used for the expression identification of the target protein. Results The optimal conditions for the induction of CRH protein were determined: temperature of 30 ℃, IPTG induced concentration 0.1 mmol/L, bacteria density (OD600) 0.8, the induction time of 8 hours, purified GST-CRH>95% fusion protein was obtained. Conclusion The optimal expression conditions of GST-CRH are obtained, and the soluble protein of high purity GST-CRH is also obtained.

Preparation of conotoxin GI antiserum and determination of neutralization activity / 军事医学

Objective To determine whether the antiserum produced by immunizing mice with conotoxin GI coupled with bovine serum albumin (BSA) could neutralize GI conotoxin.Methods The GI-BSA was prepared by glutaraldehyde-coupled method,and the mice were immunized with the GI-BSA to produce antiserum.The antibody neutralization assay was used to test the detoxication of the antiserum.Results The SDS-PAGE protein electrophoresis showed that the coupling reaction of GI hapten with BSA was successful.The two distinct protein bands of GI-BSA were more than 120×103.Each mouse was immunized four times with 99 μg every two weeks.After the fourth immunization,the serum neutralization titer was more than 1:64 000.After the intraperitoneal injection of the mixture of 100 or 200 μl of the antiserum and different doses of GI,75% of the mice survived in the group with 100 μl of the antiserum and 1× LD50 GI(16.3 μg/kg).The same percentage of mice also survived in the group of with 200 μl of serum and 25.8 μg/kg of GI.Conclusion The antiserum produced by immunizing mice with GI-BSA exhibits significant detoxication activity to conotoxin GI.

High altitude hypoxia aggravates chronic inflammation induced by LPS in rats / 军事医学

Objective To investigate the effect of high altitude hypoxia on chronic inflammation in rats.Methods Forty SD rats were randomly divided into 4 groups: control group (Con),chronic inflammation group (CI),high altitude hypoxia group (HH),high altitude hypoxia+chronic inflammation group (HH+CI).Rats in CI group were injected with lipopolysaccharide (LPS) (0.5 mg/kg) through the caudal vein twice a week for 4 weeks.Rats in HH+CI group were treated just as CI group was,but together with HH group rats were settled in a hypoxic environment of 6000 m altitude for three days.Pathological changes in lung tissues were observed by hematoxylin eosin stain.The peripheral white blood cell count and classification were measured.The levels of interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-α) in serum and lung tissues were detected by enzyme-linked immunosorbent assay (ELISA).Changes in IL-6 expression in rat lung tissues were observed by Western blotting.Results After LPS and high altitude hypoxia exposure,inflammatory cells infiltration and alveolar capillary expansion were observed in rats' lung tissue.Compared with Con group,not only the peripheral white blood cell count,but also the level of IL-6 and TNF-α in serum and lung tissue increased in CI and HH group(P<0.01).IL-6 expression levels observed by Western blotting were also increased in HH and CI group(P<0.01).High altitude hypoxia and chronic inflammation interacted(P<0.01).The peripheral white blood cell count was higher in HH+CI group than in other groups,and IL-6 and TNF-α expressions in lung tissue were increased(P<0.05).Conclusion An LPS-induced chronic inflammation model in rats is successfully obtained,and high altitude hypoxia could aggravate chronic inflammation.

Oxidative damage and related signal pathways caused by lipopolysaccharide induced sepsis in rat brain / 军事医学

Objective To investigate the mechanism of oxidative damage caused by lipopolysaccharide (LPS)induced sepsis in rat brain.Methods The rats were randomly divided into control group and model group (low LPS group and high LPS group).Twenty-four hours after the modeling,the rats were sacrificed before their brain tissue was taken out and prepared for the test.The changes in malondialdehyde (MDA),superoxide dismutase (SOD),glutathione peroxidase (GSH-Px),total antioxidant capacity (T-AOC),hydrogen peroxide (H2 O2 )and succinate dehydrogenase (SDH)were detected.The expression level of JNK and Nrf2 protein in brain tissue was detected by qRT-PCR and Western blotting. Results Compared with the control group,the MDA,SOD,GSH-px,T-AOC,H2O2 and SDH level increased significantly in the model group,and the difference in expressions of JNK and Nrf2 was statistically significant (P <0.05). Conclusion The LPS induced septic oxidative brain damage model in rats is successfully established,and the process may be regulated through the Nrf2 and JNK signal pathways.

Preparation of anti-TLR4 extracellular C-terminal domain monoclonal antibody and its application to sepsis / 中华创伤杂志

Objective To prepare the anti-TLR4 C-terminal domain monoclonal antibody and investigate its effect in treatment of sepsis.Methods TLR4 C-terminal polypeptide (amino acid sequence:368-579,named as TLR4-C) was obtained through prokaryotic expression and Sephacryl S-100 gel purification,and then was used to immunize female Balb/c mice (6-8 weeks old).After cell fusion,antibody screening and purification,monoclonal antibody specific for the C terminal of TLR4 was obtained.Specificity of monoclonal antibody was detected by Western blot and cell immunofluorescence.In vitro antibody activity test,NR8383 was cultured for 1 h with adding antibody (100 μg/ml) and then 12 h after adding lipopolysaccharide (LPS) (10 ng/ml),and level of tumor growth factor (TNF)-α in the culture medium was tested by ELISA.In vivo septic animal experiment,40 SD rats were assigned to control antibody group (n =20) and anti-TLR4 monoclonal antibody group (n =20) according to the random number table.Each group was rejected 50 mg/kg corresponding antibodies via caudal vein for 1 h,and then LPS (10 mg/kg) via intraperitoneal injection for 4 h.Blood samples from caudal vein of ten rats in each group were collect to test the serum level of TNF-α.The rest rats in each group were used to measure the animal survival rate within 72 h.Results Three highly specific anti-TLR4 monoclonal antibodies were obtained and could combined with TLR4-C and TLR4 holoprotein.In vitro cell activity study indicated only one monoclonal antibody could obviously inhibit the release of TNF-α.In vivo animal experiment showed serum TNF-α level in anti-TLR4-C antibody group was (1.54 ± 0.18) ng/ml,significantly lower than (0.51 ± 0.10) ng/ml in antibody control group (P ＜ 0.01).Animal survival rate in anti-TLR4-C antibody group was 70％,higher than 30％ in antibody control group (P ＜ 0.05).Conclusion Anti-TLR4-C monoclonal antibodies have great capacity to neutralize TLR4 and good protective effect on LPS-induced sepsis.

Stability of anticoagulant peptide Hirulog-S / 军事医学

Objective To study the stability of anticoagulant peptide Hirulog-S and its lyophilized product, and to provide data on the storage conditions and clinical applications.Methods RP-HPLC was used to determine the content and the related substances of Hirulog-S and its lyophilized powder with influence factor test, accelerated test and long-term storage test.Results Light, temperature and humidity had no significant effect on the stability of Hirulog-S and its lyophilized powder in the influence factor test.The content and related substances of Hirulog-S and its lyophilized powder did not significantly change in the accelerated test ( 40℃, RH75%) and 24-month long-term storage test at room temperature and 4℃.Conclusion Hirulog-S and its lyophilized product are very stable, even after being stored at room temperature for two years.

CXCR4 and Nrf2 expressions in non-small cell lung cancer and their clinical implications / 南方医科大学学报

<p><b>OBJECTIVE</b>To detect the expressions of CXCR4 and Nrf2 in non-small cell lung cancer (NSCLC) tissues and analyze their association with the clinicopathological features of NSCLC.</p><p><b>METHODS</b>We investigated the expressions of CXCR4 and Nrf2 in 66 NSCLC and corresponding distant normal tissue specimens using immunohistochemistry and real-time PCR.</p><p><b>RESULTS</b>The expressions of CXCR4 protein and mRNA were significantly higher in NSCLC tissue specimens than in the distant normal tissues, while the expression of Nrf2 protein and mRNA increased significantly in NSCLC tissues compared to those in the distant normal tissues (P<0.01). A high expression level of CXCR4 was positively correlated with a large tumor size (P=0.048), poor differentiation (P=0.024), advanced TNM stage (P=0.018), lymph node metastasis (P=0.004), and distant metastasis (P=0.016). The expression of Nrf2 protein was positively correlated with a large tumor size (P=0.008), advanced TNM stage (P=0.028), lymph node metastasis (P=0.038), and distant metastasis (P=0.023). A strong correlation was found between CXCR4 and Nrf2 expressions in NSCLC tissues (r=0.324, P<0.01), and the co-expression of CXCR4 and Nrf2 was strongly correlated with lymph node metastasis and distant metastasis.</p><p><b>CONCLUSION</b>Abnormal expressions of CXCR4 and Nrf2 may contribute to the progression and malignant biological behavior of NSCLC.</p>

A tracking evaluation of the quality of clinical science doctors graduating from military medi-cal colleges / 中华医学教育探索杂志

Objective To evaluate the level situation of clinical doctors graduating from mil-itary medical university within a certain period, understand their career development, and explore and analyze the problems and deficiencies in doctoral education and training to provide reference for deep-ening the reform of clinical disciplines doctoral education. Methods Stratified random samples of 9 Hospitals were selected and the established evaluation index system of clinical discipline quality tracking and investigation questionnaire was used to evaluate doctoral quality. Evaluation was surveyed by self rating and other rating combination. Quality evaluation system contains 4 first level indicators such as the military and political quality, clinical, teaching and research level, the development po-tential as well as 15 second-level indexes. The scores of self-assessment and other evaluation were compared by Pearson rank correlation analysis. The corresponding indicator scores of different doctorate types were compared using the test of variance and the subject research and published papers were analyzed by χ2 inspection. Results The comprehensive score of graduated doctors is excellent, with self-evaluation score (92.72±7.06) and other evaluation score (93.61±8.05). Correlation coefficient is 0.33(P=0.04); The academic doctors have done better in publishing papers(χ2=5.97, P=0.01) and undertaking subject research(χ2=6.08, P=0.00), but poorer in clinical work compared with the doctors of professional degree. The assessed groups are inadequate in publishing high level papers and in un-dertaking research projects and doctors of different degree types have different cognition of the ele-ments of improving the quality. Conclusions Cultivating the doctors' clinical ability and innovation ability are the cores of deepening the reform of clinical doctoral education. We should focus on inno-vation ability , value the cultivation of the clinical professional doctoral degree and explore various joint evaluation systems to attain the goal of enhancing the education quality of doctors.

Reflections on the follow-up quality survey and evaluation of the medical graduate students / 中华医学教育探索杂志

This paper analyzes the characteristics of the follow-up quality survey and evaluation of medical graduate students in China.It raises that the follow-up quality survey of graduate students in medical schools should be institutionalized,standardized and normalized; the survey and evaluation should be conducted through close cooperation with the relevant departments and training units; comparative studies on graduates of different years should be conducted; and the indicator system and survey content should value the characteristics of times,training units and subjects; Finally,specific technical issues such as when and to whom the evaluation and survey should be conducted,survey arrangements,evaluation methods,the diversity of participants and determination of results are discussed and suggestions are put forward.