ObjectiveTo observe the effects of high mobility group box-1 protein ( HMGB1 ) on function of murine spleen IL-10-producing dendritic cells (DCs) subset CD11clowCD45RBhighDCs.MethodsSplenic CD11clowCD45RBhighDCs and CD4 +T cells in Balb/c mice were purified by magnetic beads sorting.CD11clowCD45RBhighDCs were treated with various doses of HMGB1 (0,20,100,500ng/ml).Flow cytometry was used to determine expressions of CD11clowCD45RBhighDC surface molecules including CD40,CD80,CD86,Ⅰ-a/e and Toll-like receptor (TLR) 4.IL-10 level in CD11clow CD45RBhighDC culture supernatants was determined by ELISA method.The CD4+ T cells were divided into four groups: control group (without only treatment),untreated group (HMGB1-untreated CD11clow CD45RBhigh DCs plus CD4+T cells),high-dose HMGB1-treated group (500 ng/ml HMGB1-treated CD11clowCD45RBhigh DCs plus CD4 +T cells),high-dose HMGB1-treated + antibody 1 group (500 ng/ml HMGB1-treated CD11clowCD45RBhigh DCs plus IL-10 antibody and CD4+T cells),high-dose HMGB1-treated + antibody 2 group (500 ng/ml HMGB1-treated CD11clow CD45RBhigh DCs plus homotype IL-10 antibody and CD4 +T cells).IL-4 as well as interferon( IFN-γ) contents in CD4 + T cell culture supernatants were determined by flow cytometry.ResultsHMGB1 could markedly enhance the expressions of CD40,CD86 and TLR4 on CD11clowCD45RBhigh DC surface as well as level of IL-10 secreted by CD11clow CD45RBhigh DCs as compared with the non-HMGB1 treatment.Meantime,the secretion of IL-10 was HMGB1 concentration dependent.IFN-γ level in high-dose HMGB1-treated group was (279 ± 17) pg/ml,which was markedly lower than that in the untreated group [ (963 ± 11 ) pg/ml,P ＜ 0.05 \ ].IL-4 level in high-dose HMGB1-treated group was (372 ± 14) pg/ml,which was significantly higher than that in the untreated group [ (213 ± 10) pg/ml,P ＜0.05) \].ConclusionsHMGB1 promotes IL-10 production in CD11clowCD45RBhighDCs,induces the differentiation of CD4 +T cells to Th2 cells and downregulates the immune response via activating CD11clowCD45RBhigh DCs.

Objective To study the effect of Nourishing-Yin Removing-Fire and Dissipating-Stagnation Decoction on intrathyroid vascularity( ITV) and serum vascular endothelial growth factor(VEGF) levels in patients with new diagnoses of Graves disease(GD).Methods 100 patients with new diagnoses of GD were divided into two groups:Group A were treated with Nourishing-Yin Removing-Fire and Dissipating-Stagnation Decoction combined with methimazole(MT) and Group B were treated with MT alone.Thyroid volumes, superior thyroid artery peak systolic velocity(STA-PSV), thyroid vascularity index(TVI) and serum VEGF levels in patients were also observed in Group A and B before and after treatment.The correlation between thyroid volumes and serum VEGF levels in patients was analyzed.Results The parameters of GD were significantly higher than those in control group (P<0.01) .There were positive correlation between thyroid volumes and thyroid vascularity index(TVI) or serum VEGF levels in patients(r=0.57, P<0.05;r=0.59,P<0.05).The parameters in Group A and Group B decreased after treatment(P<0.01).STA-PSV, TVI and serum VEGF levels decreased significantly in Group A than those in in Group B after treatment(all P<0.05).Conclusion Nourishing-Yin Removing-Fire and Dissipating-Stagnation Decoction can improve goiters more effectively than traditional western medicine alone by decreasing serum VEGF levels of patients with GD.

Objective:To investigate the mechanism of therapeutic effects of Rhein on insulin resistance from aspects of the expression of resistin mRNA and plasma free fatty acid(FFA) in diabetic obese rats.Methods:The streptozotocin(STZ) and high fat diet-induced diabetic obese rats were divided into three groups at random.They were differently treated with Rhein,pioglitazone and saline by gavage.The expressions of resistin mRNA,plasma free fatty acid(FFA),as well as blood lipid in all groups were detected after administration by gavage for 8 weeks.Results:Similar to pioglitazone,Rhein can reduce the expression of resistin mRNA and the level of FFA,TC,TG and LDL(P

BACKGROUND: Both density gradient centrifugation and adherence method arc frequently used to isolate bone marrow mesenchymal stem cells (BMSCs).OBJECTIVE: To investigate the approaches to isolate, culture and identify the rabbit BMSCs in vitro by the combination of den,ity gradient centrifugation and adherence method. DESIGN, TIME AND SETTING: Contrast cytological study, which was performed in Central Laboratory of Shanghai 6th People's Hospital between October 2007 and March 2008.MATERIALS: Six 2-week-old rabbits were selected for BMSCs preparation and primary culture; Percoll separating medium (1.073 kg/L) was also used for this study.METHODS: BMSCs were separated and purified with Percoll separating medium by density gradient centrifugation and adherence method. The three-, five-, seven-, and nine-passage BMSCs were counted for growth curve. MAIN OUTCOME MEASURES: Morphological features and growth states of primary and passage cells were observed under inverted microscope. Indirect immunofluorescence of CD44 and CD34 antibodies were used to examine the stem cells. CD44 staining was positive, and CD34 staining was negative, suggesting the extracting and purifying cells were BMSCs. RESULTS: The passage BMSCs were uniformly distributed like fusiform shape, which were more uniform than primary cultured cells. The BMSCs grew productively and proliferated rapidly; meanwhile, the nucleolus was clear, caryopla.sm was in a large proportion, morphological features were uniform, ceils like bostrychoid or whirlpool were arranged parallelly, and the five-pa.ssage cells were not changed remarkably. Proliferation was decreased gradually with the passage increasing; especially, the proliferation of three-five-passage cells was the strongest. The separated cells expressed CD44 but not CD34. CONCLUSION: High-purified rabbit BMSCs are obtained by both density gradient centrifugation and adherence method.

Objective:To determine the effects of TPS on peripheral blood Tregs in sepsis mouse induced by burn plus P.aeruginosa infection.Methods: The experimental mice were separated into five groups randomly ,including sham burn group ,burn plus P.aeruginosa infection group ,burn plus P.aeruginosa infection with TPS (50,100,200 mg/kg) treatment group.Peripheral blood Tregs were isolated with Magnetic Microbeads and cultured in vitro from the day after burn (PBD0) to 4 days after burn(PBD4).IL-10, IFN-γ,IL-4 levels in Tregs culture supernatants were determined by sandwich enzyme-linked immunosorbent assays ( ELISA ) . Purification of CD4+CD25high Tregs and CD4+T cells in C57BL/6 mice were administrated by magnetic beads sorting .Tregs and CD4+T cells were cultured in vitro after joining TPS to without TPS cells as a control .The phenotypes of Tregs were analyzed by flow cytometry , and cytokines were measured by ELISA .Results:Vis-a-vis the results of the untreated group ,TPS could markedly decrease IL-4 and IL-10 secretion level and significantly increase the secretion of IFN-γ,and the secretion of IL-10 level and concentration of TPS dose effect.Vis-a-vis the results of the untreated group ,in vitro experiment ,without stimulation of TPS ,CD4+T cell proliferation and IFN-γwere significantly reduced ( P<0.05 ) and IL-4 levels increased significantly;CD4+T cell proliferation and IFN-γwere significantly increased and IL-4 levels were significantly reduced in the group of TPS with antibody-1;there was no significant difference in CD 4+T cell proliferation and the levels of IFN-γand IL-4 in the group of TPS with antibody-2.Conclusion:TPS could inhibit the abnormal ac-tivities of CD4+CD25highTregs in burn with P.aeruginosa infection mice,at least in part via inhibiting IL-10 secretion,and trigger a shift of Th2 to Th1 with activation of CD4+T cells in burn with P.aeruginosa infection mice.

Objective To observe the effects of electro-acupuncture (EA) on body weight, insulin resistance, hypothalamic agouti gene-related protein (AGRP) and neuropeptide Y (NPY) in diet induced obesity (DIO) rats; To discuss its mechanism for DIO.Methods Fifty SD male rats were randomly divided into low fat diet groups (LFD) and high fat diet group. After the DIO models were established, the successful model rats were randomly divided into model group, EA group and Orlistat (OLST) group. EA was applied to “Zusanli” (ST36) and “Quchi” (LI11) for 20 minutes. The treatment was done once a day for 28 days. OLST was treated with orlistat by gavage. LFD and model did not receive treatment. Body weight was recorded every day. FPG and FINS were detected. The expressions of AGRP and NPY were detected by RT-QPCR. Morphological changes of adipocyte and liver were examined by HE staining.Results The body weight, HOMA-IR, AGRP, NPY and diameter of adipocyte of EA group and OLST group were significantly lower than model group (P<0.05), difference between EA group and OLST group. The states of hepatic steatosis were improved in EA group and OLST group.Conclusion EA has the effects on weight loss by inhibiting the expressions of AGRP and NPY by improving IR.

Background Diabetic retinopathy (DR) has become the main cause of blindness in the world,But the etiology of DR is still not clear,and the results of the studies on the risk factors of DR are not completely consistent.Fully understanding the risk factors of DR has an important clinical value for the prevention and treatment of DR.Objective This study was to analyze the prevalence rate and risk factors of DR inpatients with type 2 diabetes and provide a basis for the establishment of ophthalmic intervention programs and measures for diabetic inpatients.Methods Cross sectional study was performed.Four hundred and seventy three patients with type 2 diabetes in the department of endocrinology,Dongguan People's Hospital from July 2011 to July 2012 were included.The patients were divided into DR group and non diabetic retinopathy (NDR) group.The DR group was subdivided into mild,moderate and severe non-proliferative DR (NPDR) group and proliferative DR (PDR) group.The sex and age of patients,course of diabetes,body mass index (BMI),systolic blood pressure,diastolic blood pressure,fasting blood glucose (FBG),2 hours postprandial blood glucose (2 h PBG),glycosylated hemoglobin (HbA1 c),fasting insulin,2 hours postprandial insulin,high density lipoprotein cholesterol,low density lipoprotein cholesterol,apolipoprotein A1 (APOA1),APOB,alpha lipoprotein,total cholesterol,three glycerol,urea nitrogen,creatinine,uric acid,percentage of neutrophil,24 hours urinary albumin total (ALBU-24 h) and ALBU were detected.Logistic regression was used to analyze the relationship between DR and various factors,and the risk factors of DR were screened out.Results The prevalence of DR inpatients with type 2 diabetes was 28.33％,the prevalence of mild,moderate and severe NPDR were 2.54％,16.28％ and 4.23％,respectively,the prevalence of PDR was 5.29％.The prevalence of DME was 10.36％ in the DR patients.The course of diabetes,the levels of serum lipoprotein,creatinine,ALBU-24 h and ALBU were statistically significant between DR group and NDR group (all at P＜0.05).By stepwise Logistic regression analysis,the course of disease,FBG were identified as the independent risk factors of DR (course of disease:odds ratio [OR] =1.155,95 ％ confidence interval [CI]:1.067-1.251;FBG:OR =1.313,95％ CI:1.071-1.610).Conclusions The course of diabetes,lipoprotein,creatinine,ALBU-24h,ALBU are closely related to the occurrence and development of DR.The course of diabetes and FBG are the independent risk factors of DR.

Abstract: Objective　To investigate the molecular characteristics and drug resistance of Staphylococcus aureus (SA) isolated from wounds of paatients with orthopedic trauma, and analyze the molecular subtyping, virulence genes and drug resistance of SA in wounds of patients, so as to provide reference for the prevention and treatment of wound SA infection in patients. Methods　From January 2020 to June 2022, a total of 128 SA isolates were collected from wound specimens of orthopedic trauma patients at Wuxi 9th People's Hospital Affiliated to Soochow University. Methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-susceptible Staphylococcus aureus (MSSA) were differentiated using PCR. Multilocus Sequence Typing (MLST), staphylococcal protein A (spa), staphylococcal chromatoidal cassette mec (SCCmec), and accessory gene regulator (agr) typing were performed to determine the molecular typing and presence of virulence genes and drug resistance profiles. Results　Among the 128 SA isolates, 76 (59.38%) were MRSA and 52 (40.62%) were MSSA. MRSA typing showed that, MLST was dominated by ST59 (46 strains, 60.53%), spa was dominated by t437 (52.63%), SCCmec was dominated by Ⅰ (42.11%) and Ⅳ (39.47%). MSSA typing showed that, MLST was dominated by ST188 (30.77%), spa was dominated by t189 (61.54%), agr was dominated by Ⅰ (53.85%). In MLST typing, ST59 of MRSA was higher than that of MSSA, and ST188 and ST6 of MRSA were lower than those of MSSA (χ2=36.207, 20.227, 9.984, P<0.05). In spa typing, the t437 of MRSA was higher than that of MSSA, and the t189 of MRSA was lower than that of MSSA (χ2=18.276, 32.781, P<0.05). The virulence genes showed that, the detection rates of hlb and seb in MRSA were higher than those in MSSA (χ2=47.838, 10.261, P<0.05), and the detection rates of cna and ebpS in MRSA were lower than those in MSSA (χ2=26.176, 8.305, P<0.05). Drug susceptibility test showed that, and the drug resistance rates of MRSA and MSSA to vancomycin (VAN) and linezolid (LNZ) were 0. The drug resistance rates of MRSA to oxacillin (OXA), ERY and CLI were 86.84%, 68.42% and 76.32%, which were higher than corresponding 7.69%, 42.31% and 46.15% of MSSA (χ2=78.055, 8.623, 12.200, P<0.05). The analysis of multi-drug resistant strains (MDR) showed that 76 MRSA strains were MDR strains, and 12 of 52 MSSA strains (23.08%) were MDR strains. Conclusions　The molecular characteristics of SA isolated from orthopedic trauma patients' wounds were predominantly associated with MRSA strains of ST59-t437-SCCmec Ⅰ/Ⅳ-MRSA and ST188/ST6-t189-agr Ⅰ. These strains showed higher resistance to oxacillin, erythromycin, clindamycin, and higher susceptibility to vancomycin and linezolid. Such characteristics were closely related to the carriage of virulence genes. Clinicians should pay attention to the presence of MDR MSSA and develop appropriate antimicrobial strategies based on SA's molecular characteristics and antimicrobial resistance.

OBJECTIVETo discuss the operation method and characteristic of correcting upper eyelid depression by transposition of orbital septum fat.

METHODSDuring the double eyelid surgery, we set.the lateral orbital septum fat completely free, while the bottom is still connected with the middle orbital septum fat. We separate a tunnel from the middle to the inner side in orbital septum, and the separated orbital septum fat is transposed to the inner side of orbital septum by the tunnel with suturing fixation.

RESULTSFrom March 2008 to October 2013, 51 cases with upper eyelid depression were treated successfully. Patients were followed up for 3 months to 3 years (average, 7. 5 months) with sustained aesthetic results.

CONCLUSIONSOrbital septum fat transposition can successfully correct the upper eyelid depression. It should become a regular procedure in blepharoplasty.

Adipose Tissue ; transplantation ; Blepharoplasty ; methods ; Esthetics ; Eyelids ; abnormalities ; surgery ; Humans ; Orbit

Objective To investigate the bcl?2 gene modification on neurological function recovery in rats with spinal cord injury in neural stem cell transplantation. Methods Cultured rat neural stem cells by Ad?EGFP as vector?mediated side B?cell lymphoma 2 gene ( bcl?2 ) gene transfection of neural stem cells were divided into 3 groups: control group, negative transfection group, bcl?2 transfection group. Use western?blot to detect the expression of bcl?2 protein in neural stem cells before and after transfection. 85 adult female SD rats, successful model 72, were randomly divided into control group, NSCs group, bcl?2?NSCs groups, 24/group, rat acute spinal cord injury model in accordance with a modified Allen’ s method. Assess the motor function by BBB rating and the swash plate test. 7 days after modeling by RT?PCR and Western blot detection of spinal cord injury around HSP27, c?fos gene expression, TUNEL assay apoptosis. Four weeks after model drawn line HE staining and fluorescence microscopy EGFP?labeled NSC survival and distribution of the rats neurophysiological recovery by SEP and MEP. Results bcl?2 gene transfection of rat neural stem cells, bcl?2 transfection group and control group, negative transfection group compared to bcl?2 mRNA and protein levels were expressed ( P < 0. 05 ); lower extremity motor function in rats evaluation of bcl?2?NSCs group than NSCs group, NSCs group than the control group. 72 hours after modeling, bcl?2?NSCs number of apoptotic cells were significantly lower than the control group and NSCs group (P < 0. 05). 7 days after modeling, compared with the control group and NSCs group, bcl?2?NSCs group HSP27 gene and protein expression was significantly higher than that (P < 0. 05), bcl?2?NSCs group c?fos mRNA and protein expression was significantly reduced compared (P < 0. 05). 4 weeks after modeling, HE staining control group showed spinal cord tissue loss and the formation of syringomyelia, no axonal through. NSCs group damage zone few of neuraxis?like structures, syringomyelia smaller, bcl?2?NSCs group showed more nerve axon?like structure, no syringomyelia. EGFP?positive cells labeled:bcl?2?NSCs group the most, NSCs group followed, no control group, and the difference between the groups was statistically significant (P < 0. 05). After the 4th week, SEP and MEP latency period:bcl?2?NSCs group NSCs group > control group, and between the groups was significant difference ( P < 0. 05 ) . Conclusions By Ad?EGFP as vector?mediated side B?cell lymphoma 2 gene (bcl?2) gene transfection make neural stem cells can promote cultured rat neural stem cells. bcl?2 gene?modified neural stem cell transplantation can promote the regeneration of spinal cord injury synaptic elevated HSP27 expression after spinal cord injury, reduced expression and neural cell apoptosis after spinal cord injury bcl?2 gene and improve limb movement in rats function and electrophysiological function.