Objective To investigate the effect of δ-opioid receptor agonist D-Ala2-D-Leu5-enkephalin (DADLE) on the celluar immune function in septic rats. Methods One hundred and fifty healthy male SD rats weighing 154-198 g were randomly divided into 3 groups (n = 50 each):group Ⅰ sham operation (group S);group Ⅱ sepsis (group SEP) and group Ⅲ DADLE. Sepsis was induced by cecum ligation and punture (CLP) in group Ⅱ and Ⅲ. In group Ⅲ 0.5 rmg/kg DADLE 10 ml/kg was injected intraperitoneally (IP) immediately after CLP operation. Seven day survival rate was calculated. Blood samples were collected from 10 animals at 4, 8 and 12 h after operation (T1-3) respectively in each group for determination of serum TNF-α and IL-10 concentrations by ELISA and changes in T-cell subsets by flow cytometry. Results CLP significantly increased serum TNF-α and IL-10 concentrations and TNF-α/IL-10 ratio at T1-3 and decreased CD4+ and CD4+/CD8+ ratio and increased CD8+ at T3 in group Ⅱ as compared with group S (group Ⅰ). DADLE treatment significantly attenuated the CLP-induced above changes. Seven-day survival rate was significantly higher in DADLE group than in CLP group.Conclusion δ-opioid receptor agonist DADLE can improve the celluar immune function of rats with sepsis and increase the survival rate.

Objective To investigate the characteristic of NK cells and NKT cells in HBV infected pediatric subjects for evaluation of their clinical implication. Methods Fresh peripheral blood samples were obtained from 42 HBV-infeeted pediatric cases and 15 healthy counterparts. NK cells and NKT cells were analyzed by flow cytometry assay. The ehnical data such as serum ALT level and HBV viral load was simultaneously recorded from each HBV cartier. Results HBV-infected children had an obviously increasing percentage of NK cells 12.071% ± 7.100%, there were significant differences between the children with chronic B hepatitis and the healthy children ( P < 0.05). As far as percentage of NKT 3.048% ± 1.937% was concerned, there were not differences. Furthermore the association was not found between serum HBV viral load level and the NK lymphocyte. Conclusion Our data may provide valuable information of NK .and NKT lymphocyte for evaluation of disease progression of HBV infected children NKI cells.

Objective To explore clinical efficacy of F-type forceps for assisted reduction in femoral shaft fracture reduc-tion.Methods Forty-five patients with femoral shaft fracture treated with intramedullary nail and internal fixation from January 2019 to December 2021 were retrospectively analyzed and divided into two groups according to different reduction methods.In observation group,there were 21 patients,included 15 males and 6 females,aged from 27 to 92 years old with an average of(53.38±18.81)years old;9 patients on the left side,12 patients on the right side;7 patients were type A,8 patients were type B and 6 patients were type C according to AO fracture classification;the time from injury to operation ranged from 7 to 13 days with an average of(4.62±3.34)days;reduction was assisted by F-shaped forceps.In control group,there were 24 patients,in-cluding 17 males and 7 females,aged from 20 to 92 years old with an average of(51.96±20.43)years old;12 patients on the left side,12 patients on the right side;11 patients were type A,8 patients were type B and 5 patients were type C according to AO fracture classification;the time from injury to operation ranged from 2 to 13 days with an average of(6.29±3.04)days;tra-ditional reset mode was adopted.Operative time,intraoperative blood loss,intraoperative fluoroscopy times,intraoperative open reduction ratio,clinical healing time of fracture,postoperative complications,hospital stay,hospital cost and Lysholm score of knee joint at 6 and 12 months after surgery were compared between two groups to evaluate clinical effect.Results All patients were followed up for 12 to 24 months with an average of(16.60±3.45)months.In observation group,operative time,intraoper-ative blood loss,intraoperative fluoroscopy times,open reduction cases,and clinical healing time of fractures were(58.19±7.93)min,(88.10±44.45)ml,(25.29±5.54)times,0 case,(4.76±0.77)months,respectively;while in control group was(79.33±22.94)min,(222.92±144.45)ml,(47.46±26.25)times,5 cases,(7.13±1.80)months,and the difference between two groups were statistically significant(P＜0.05).There were no significant difference in postoperative complications,length of stay and hospitalization cost between two groups(P＞0.05).At 6 months after surgery,Lysholm score of knee joint in observa-tion group(88.62±4.48)was better than that in control group(79.21±8.91)(F=21.948,P=0.000).There were no significant difference in support use,pain and squat score between two groups(P＞0.05).At 12 months after surgery,Lysholm scores of stair climbing and pain in observation group were(9.62±1.20)and(19.76±1.92),which were better than those in control group(7.83±2.04)and(21.88±2.88)(P＜0.05).There were no significant difference in scores and total scores of other items between two groups(P＞0.05).Conclusion Compared with traditional reduction method,F-type forceps instrument could short-en operation time,reduce intraoperative blood loss,reduce intraoperative fluoroscopy times,accelerate clinical healing of frac-ture,and promote earlier functional recovery of knee joint.

Human glutamate decarboxylase 65(hGAD65) is an enzyme that catalyzes the transformation of L-glutamic acid into ?-aminobutyric acid.It has been found that Type 1 diabetes mellitus(T1DM)is an autoimmune disease,in which pancreatic islet ?-cells are destroyed due to immune response mediated by autoantigen.hGAD65 is considered as a key autoantigen of the autoimmune response,so anti-hGAD65 antibody(hGAD65-Ab) is the most effective and specific immune marker for T1DM diagnosis,and hGAD65 can be used to detect hGAD65-Ab in serum of T1DM patients.The hGAD65 gene was cloned into pET32a(+),then the recombinant plasmid with hGAD65 was transformed into E.coli BL21(DE3) and expressed by IPTG induction.The fusion protein containing thioredox,hexahistidine and hGAD65(Trx-hGAD65) was mostly insoluble,but the band of soluble Trx-hGAD65 could also be detected by SDS-PAGE,and it was a great improvement compared with the results reported.Trx-hGAD65 was isolated from lysate and purified by immobilized metal ion affinity chromatography(IMAC).After enterokinase digestion and IMAC purification,hGAD65 with high purity was obtained.Detection of thin-layer chromatography(TLC) showed that both Trx-hGAD65 and hGAD65 had enzymatic activity,whereas Trx-hGAD65 had better stability.Furthermore,it was confirmed that Trx-hGAD65 was able to conjugate with hGAD65-Ab in the serum of T1DM patients by ELISA assay.In conclusion,Trx-hGAD65 instead of hGAD65 can be used for T1DM diagnosis,and its application in prophylaxis and therapy of T1DM is expectable.

Objective To study the influences of mental disorders on female systemic lupus erythematosus(SLE)and analyze the factors. Methods We used symptom check list -90 (SCL-90) as a basis for judging mental disorders disease activity. Disease activity, social support and depreciation - discrimination were used as possible influencing factors. Social support and discomfort – discrimination were possible influencing factors. Multivariate unconditional logistic regression model was used to analyze the influencing factors of mental disorders. Results The total score of SCL-90 of patients with female SLE was significantly higher than that of norm models ［(136.39±48.66) vs (129.96±38.76)］ (P<0.05), in 289 SLE patients, the number of patients with mental disorders was 128 (44.3%). High monthly income(OR=0.770, 95% CI:0.604-0.981, P=0.034) was a protective factor for mental disorders. High disease activity (OR=1.792, 95% CI:1.023-3.138, P=0.042)and high discomfort–discrimination (OR=1.100, 95% CI:1.035-1.169, P=0.002)were risk factors for mental disorders. Conclusions Female SLE patients have a higher risk of mental disorders than the general population. And eliminating self-depreciation, reducing social discrimination, active employment, increasing monthly income, standardizing treatment and reducing disease activity may effectively alleviate mental disorders in SLE patients.

ATP-Binding Cassette, Sub-Family B, Member 1 ; analysis ; Cytotoxicity, Immunologic ; Dendritic Cells ; physiology ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Humans ; Immunophenotyping ; K562 Cells ; Leukemia ; drug therapy ; immunology ; Lymphocyte Culture Test, Mixed

To investigate the effects of 2-(4-methoxycarbonyl-2-tetradecyloxyphenyl)carbamoylbenzoic acid (CX09040) on protecting pancreatic β cells, the β cell dysfunction model mice were induced by injection of alloxan into the caudal vein of ICR mice, and were treated with compound CX09040. Liraglutide was used as the positive control drug. The amount and the size of islets observed in pathological sections were calculated to evaluate the β cell mass; the glucose stimulated insulin secretion (GSIS) test was applied to estimate the β cell secretary function; the oral glucose tolerance test (OGTT) was taken to observe the glucose metabolism in mice; the expressions of protein in pancreas were detected by Western blotting. The effects on the target protein tyrosine phosphatase 1B (PTP1B) were assessed by the PTP1B activities of both recombinant protein and the intracellular enzyme, and by the PTP1B expression in the pancreas of mice, separately. As the results, with the treatment of CX09040 in alloxan-induced β cell dysfunction mice, the islet amount (P<0.05) and size (P<0.05) increased significantly, the changes of serum insulin in GSIS (P<0.01) and the values of acute insulin response (AIR, P<0.01) were enhanced, compared to those in model group; the impaired glucose tolerance was also ameliorated by CX09040 with the decrease of the values of area under curve (AUC, P<0.01). The activation of the signaling pathways related to β cell proliferation was enhanced by increasing the levels of p-Akt/Akt (P<0.01), p-FoxO1/FoxOl (P<0.001) and PDX-1 (P<0.01). The effects of CX09040 on PTP1B were observed by inhibiting the recombinant hPTP1B activity with IC50 value of 2.78x 10(-7) mol.L-1, reducing the intracellular PTP1B activity of 72.8% (P<0.001), suppressing the PTP1B expression (P<0.001) and up-regulating p-IRβ/IRβ (P<0.01) in pancreas of the β cell dysfunction mice, separately. In conclusion, compound CX09040 showed significant protection effects against the dysfunction of β cell of mice by enlarging the pancreatic β cell mass and increasing the glucose-induced insulin secretion; its major mechanism may be the inhibition on target PTP1B and the succedent up-regulation of β cell proliferation.
Alloxan ; Animals ; Benzoates ; pharmacology ; Biological Assay ; Disease Models, Animal ; Glucose ; metabolism ; Glucose Tolerance Test ; Insulin ; secretion ; Insulin Resistance ; Insulin-Secreting Cells ; drug effects ; Liraglutide ; pharmacology ; Mice ; Mice, Inbred ICR ; Molecular Weight ; Pancreas ; drug effects ; enzymology ; Protein Tyrosine Phosphatase, Non-Receptor Type 1 ; antagonists & inhibitors ; Signal Transduction

Objective To construct a new conditionally replicative adenovirus (CRAds) targeting carcinoembryonic antigen (CEA) positive colorectal cancer cells. Methods The DNA fragment of the CEA gene promoter was amplified through PCR and cloned into the vector carrying fusion reporter gene EGFP-Luc to construct expression plasmid pCEA-EGFPLuc. The constructed plasmid pCEA-EGFPLuc was transfected into CEA positive and negative cells by liposome. The activity of CEA gene promoter was evaluated by detecting the expression of EGFP and luciferase activity. The conditionally replicative adenovirus Ad.CEA-E1A/CMV-TK carrying suicide gene HSVtk was constructed, in which the E1A gene was controlled under CEA promoter. CEA positive(Lovo and SW620)and negative tumor cells(HeLa) were infected with Ad.CEA-E1A/ CMV-TK. The selective cytotoxicity of Ad.CEA-E1A/CMV-TK and the synergistic effect of the virus with GCV in CEA positive tumor cells were evaluated by the expression of E1A, cytopathic effect and cell survival rate. Results CEA promoter possesses a good specificity as well as high activity. The expression of E1A only presented in CEA positive tumor cells. After infection with Ad. CEA-E1A/CMV-TK, the cell survival rates of Lovo and SW620 were (36.72±2.49)% and (39.82±4.76)%, respectively, significantly lower than that of Hela[(87.44±2.76)%1 (P<0.01). When combined with GCV, Ad.CEA-E1A/CMV-TK had better oncolytic effect on Lovo and SW620 cells, with cell survival rates of (17.26±3.65) % and (23.93±5.40) %, respectively, significantly lower than those without GCV[(36.72±2.49) % and (39.82±4.76) %, respectively] (P<0.01). Conclusion Ad. CEA-E1A/CMV-TK under the control of CEA promoter has selective cytotoxic effect on CEA positive colorectal cancer cells, and the effect can be enhanced when combined with GCV.

We conducted prediction and analysis for secretory proteins from Thelazia callipaeda at Genome Scale based on the previous full genome annotation.The software SignalP,TMHMM,big-PI Predictor,MEME,Protcomp and SecretomeP were combined to process the prediction of the secretome of Thelazia callipaeda.The analyses of secretory proteins by GO function enrichment,KEGG pathway,and statistics of domains were performed.Results showed that totally 259 secretory proteins were found in Thelazia callipaeda genome and the amino acid lengths of secretory proteins were mainly concentrated between 100 to 700 aa exclusively.GO function analysis of secretory proteins indicated that they were enriched in the secreting pathways and in the interactions with host.The results of KEGG metabolism secretory proteins suggested that some of them contributed to drug metabolism and glutathione metabolism.And domain analysis suggested that most of them were glycoside hydrolase,contributing to sugar metabolism.Around 126 secretory proteins had antigenicity of B-cell epitope.In summary,we found that secretory proteins in Thelazia callipaeda were most small proteins,which were involved in sugar metabolism and antioxidative activity,facilitating Thelazia callipaeda to invade the hosts and play a key role in the parasitic course.

OBJECTIVETo explore the safety and effectiveness of endoscopic mucosal resection (EMR) in the treatment of colorectal polyps.

METHODEMR was applied in the treatment of colorectal polyps.

RESULTSA total of 3578 polyps in 2609 patients were all completely resected except 2 cases and the integrated rate of samples was 99.6%. Intra- and post-operation complications occurred in 22 cases(0.8%), including 7 intraoperative bleeding, 5 postoperative bleeding, and 10 thermal burn, which were cured by symptomatic treatment. A total of 1530 (58.6%) cases were followed-up with 3-12 months and no relapse was found in former place of excision.

CONCLUSIONEMR can be applied in resection of colorectal polyps effectively and safely.