Objective: To investigate the influence of transfection of TCRVJ37.1 gene in cytotoxicity of PBMC of healthy donor to breast cancer cell line MCF-7/S. Methods :pcDN*31vp'1 packaged by lipofectamine were transfected to healthy donor's PBMC,Flow Cytometer Analysis and MTT Colorimetric Assay were used respectively to test the expression of pcDNA3.1VB7.1 gene and the cytotoxicity of PBMC before and after gene transfection of healthy donor to breast cancer cell line MCF-7/S. Results: The expression of TCRVJ37.1 gene after transfection was obviously higher.There was distinguish differences of cytotoxicity before and after gene transfection. Conclusion:The modified healthy donor's PBMC by TCR gene could have stronger cytotoxicity to breast cancer cell line.

Objective To investigate the effect of vascular endothelial growth factor (VEGF) on the leukocyle function in a rat denervated infect flap model and to investigate the action mechanison of VEGF in the flap anti-infection .Methods An island pedicle flap measured 2 cm × 2 cm was raised on the right abdomen of sixty wister rats ,which were divided into three groups .All flaps re-ceived intradermal inoculation of 107 Staphylococcus aureus ,and the animals were observed for 96 h .The indexes of the leukocyte count ,leukocyte vitality ,hemiluminescence of neutrophils ,tissue bacterial count ,naked eyes observation of falptissue and the light microscope observation were detected .Results The leukocyte count in the exudation at postoperative 96 h had no statistical differ-ences among 3 groups(P>0 .05);while in the indexes detection of the leukocyte vitality ,chemiluminescence of neutrophils ,tissue bacterial count ,etc .,the comparison between the chronic denervated group and the control group showed very significantly differ-ences(P<0 .01);the differences between the VEGF treatment group and the chronic denervated group was very significant (P<0 .01) .The falp pathological change in the control group and the VEGF treatment groups was slight .Conclusion The soft issue af-ter losing the innervation decreases the leukocyte function .VEGF might improve the flap micro circulation and play an important role in improving the leukocyte function .

AIM: To investigate the effects of fibroblast growth factor 10 ( FGF10 ) on lipopolysaccharide ( LPS)-induced microglial activation .METHODS:Mouse BV2 microglial cells were maintained in DMEM in a humidified incubator with 95%/5%( V/V) mixture of air and CO 2 at 37℃.The medium was changed every 1 or 2 d.The cells were digested and passaged every 4 or 5 d.The BV2 microglial cells were first pretreated with FGF 10 (1 mg/L) for 30 min and then stimulated with LPS (500 μg/L).The medium and the cells were collected at different time points .The morphologi-cal changes of microglia were visualized under microscope .To evaluate the microglial activation , the transcription and pro-duction of proinflammatory factor tumor necrosis factor-α( TNF-α) were examined by real-time quantitative polymerase chain reaction (RT-qPCR) and enzyme-linked immunosorbent assay (ELISA), respectively.RESULTS:The morphology of control BV2 microglia showed circular or oval shape .After exposure to LPS for 24 h, the microglia revealed spindle shaped or multipolar morphology , and the percentage of activated cells was significantly increased compared with control group.Pretreatment with FGF10 successfully inhibited the morphological change from normal to activated shape .LPS sti-mulation for 6 h significantly increased the transcription of TNF-α, while FGF10 pretreatment remarkably reversed the effect.In addition, the production of TNF-αincreased in the presence of LPS stimulation for 24 h compared with control group.Pretreatment with FGF10 suppressed LPS-induced TNF-αexpression.CONCLUSION: Pretreatment with FGF10 inhibits the morphological change from normal to activated shape , and remarkably suppressed the transcription and produc-tion of TNF-α.FGF10 successfully suppresses LPS-induced BV2 microglial activation , indicating that FGF10 is a therapeu-tic agent for the treatment of glia-mediated neuroinflammatory diseases .

Systemic sclerosis (SSc) is an autoimmune disease characterized by thickening of the skin and organ fibrosis.Ankylosing spondylitis (AS) is a type of arthritis with long-term inflammation of the axial joints.Previous studies presented 5 cases of concomitant AS and SSc.However,there was only 1 patient of those 5 cases complaining of muscle weakness while all patients had approximately normal creatine kinase (CK).Here we reported a young male who met the criteria for SSc and AS while showing significantly elevated CK.Human leukocyte antigen (HLA) typing results indicated the genetic susceptibility to these two diseases.The patient was prescribed prednisone (30 mg/d)and cydophosphamide.After 2 months,the patient's skin became soft with normal CK.

Molecular mechanisms underlying the effects of Fyn on cell morphology, pseudopodium movement, and cell migration were investigated. The Fyn gene was subcloned into pEGFP-N1 to produce pEGFP-N1-Fyn. Chinese hamster ovary (CHO) cells were transfected with pEGFP-N1-Fyn. The expression of Fyn mRNA and proteins was monitored by reverse transcription-PCR and Western blotting. Additionally, transfected cells were stained with 4',6-diamidino-2-phenylindole and a series of time-lapse images was taken. Sequences of the recombinant plasmids pMD18-T-Fyn and pEGFP-N1-Fyn were confirmed by sequence identification using National Center for Biotechnology Information in USA, and Fyn expression was detected by RT-PCR and Western blotting. The morphology of CHO cells transfected with the recombinant vector was significantly altered. Fyn expression induced filopodia and lamellipodia formation. Based on these results, we concluded that overexpression of mouse Fyn induces the formation of filopodia and lamellipodia in CHO cells, and promotes cell movement.
Animals ; Blotting, Western ; CHO Cells ; Cricetinae ; Cricetulus ; Genetic Vectors ; Green Fluorescent Proteins/genetics ; Mice ; Proto-Oncogene Proteins c-fyn/genetics/*metabolism ; Pseudopodia/*metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Time-Lapse Imaging ; Transfection

AIM:To investigate the effect of microRNA-214 ( miR-214) on cardiomyocyte hypertrophy and the expression of the potential target genes .METHODS:A cell model of hypertrophy was established based on angiotensin-Ⅱ( Ang-Ⅱ)-induced neonatal mouse ventricular cardiomyocytes ( NMVCs) .Dual luciferase reporter assay was performed to verify the interaction between miR-214 and the 3’ UTR of MEF2C.The expression of MEF2C and hypertrophy-related genes at mRNA and protein levels was determined by RT-qPCR and Western blot , respectively .RESULTS:The expression of ANP, ACTA1,β-MHC and miR-214 was markedly increased in Ang-Ⅱ-induced hypertrophic cardiomyocytes .Dual lu-ciferase reporter assay revealed that miR-214 interacted with the 3’ UTR of MEF2C, and miR-214 was verified to inhibit MEF2C expression at the transcriptional level .The protein expression of MEF2C was markedly increased in the hypertro-phic cardiomyocytes .Moreover, miR-214 mimic, in parallel to MEF2C siRNA, inhibited the expression of hypertrophy-re-lated genes in Ang-Ⅱ-induced NMVCs.CONCLUSION:MEF2C is a target gene of miR-214, which mediates the effect of miR-214 on attenuating cardiomyocyte hypertrophy .

Objective:To explore the debridement effectiveness of infected bone tissue of chronic hematogenous osteomyelitis in the lower extremities under the guidance of 99mTc-MDP SPECT/CT fused images. Methods:A retrospective case series analysis was conducted on 21 patients with chronic hematogenous osteomyelitis in the lower extremities treated at Southwest Hospital of Army Medical University from May 2017 to June 2020. There were 8 males and 13 females, with the age range of 10-62 years [23(18, 37)years]. The tibial infections were found in 16 patients, and femoral infections in 5 patients. The duration of bone infection was 4-480 months [120(42, 228)months]. According to the Cierny-Mader anatomico-physiological system, 4 patients were classified as type I, 14 as type III, 3 as type IV; 18 patients were classified as type A and 3 as type B. Intraoperative debridement of infected bone tissue was operated at stage I on the region of interest (ROI) where the isocontour(ISO) value was between 30%-40%, using the preoperative 99mTc-MDP SPECT/CT fused images as the reference. The stage II bone defect reconstruction was based on autologous and / or allogeneic bone. To observe the frequency of operations regarding bone infection control in stage I. The preoperative white blood cell count (WBC), erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), intraoperative bacterial culture and pathological examination were compared at stages I and II. The skin redness and swellings, pain, sinus tract in the infected limbs, and ossification of grafted bones in the original bone defect part were observed at stage II. The accuracy rate between ISO value in the region of interest (ROI) and set ISO figure was checked. The difference of longitudinal length of the bone debridement area in ROI area with the actual bone debridement area was observed under the coronal position. Results:All patients were followed up for 6-36 months [11(9, 29)months] after stage II operation. All of the 21 patients had undergone operations of infection control with an average number of 1.04 times in stage I. 1 patient's intraoperative frozen section indicated that neutrophils were>5/HP. The bone graft at stage II had been completed after another debridement. Comparison of preoperative inflammatory markers at stages I and II: the WBC was decreased from (5.9±1.6)×10 9/L to (5.4±1.5)×10 9/L ( P>0.05), the ESR decreased from 9(5, 26)mm/h to 4(2, 10)mm/h ( P<0.05), and the CRP decreased from 2.8(2.3, 7.7)mg/L to 2.3(1.4, 3.0)mg/L ( P>0.05). The results of bacterial culture of tissue at stage I were positive in 12 patients and negative in 9 patients. The pathological examination indicated neutrophils and lymphocyte infiltration. The results of bacterial culture of tissue at stage II were all negative. A modicum of plasmacyte and lymphocyte infiltration and the neutrophils (<5 per/Hp) had been found in the intraoperative frozen section and pathological examination. No redness, swelling or sinus tract was found in the skin after stage II surgery and ossification of grafted bone was good. The accuracy rate between ISO value in the ROI and set ISO figure was 90.5%. The comparison between longitudinal debridement scope of ROI [(86.8±31.1)mm] and actual bone tissue debridement scope [(86.0±31.3)mm] at stage I showed no significant difference ( P>0.05). Conclusions:99mTc-MDP SPECT/CT fused images can be used as an effective means to define the debridement scope of infected bone tissue preoperatively. The method can not only avoid excessive debridement, but also improve the cure rate of hematogenous osteomyelitis in the lower extremities.

To investigate the clinical characteristics and prognosis for connective tissue disease (CTD) with cryptococcal meningitis (CM). 
 Methods: Clinical data of 18 patients with CTD complicated with cryptococcal meningitis diagnosed by Rheumatology and Immunology Department, Xiangya Hospital, Central South University from January 2000 to January 2017, were retrospectively analyzed.
 Results: The common symptoms of CTD patients with CM were headache, fever, nausea, and vomiting. Patients with severe clinical manifestations, such as convulsions and disturbance of consciousness, all died. Logistic regression analysis showed that disturbance of consciousness and decreased peripheral blood lymphocyte count might be the related factors of poor prognosis of CTD patients with CM (P<0.05). The mortality rate of CTD with CM was 61.11%, and the effective rate of treatment for this disease was 38.89%.
 Conclusion: CTD patients with cryptococcal meningitis have a high risk of death. Severe clinical symptoms, such as disturbance of consciousness and lower peripheral blood lymphocyte count, are associated with its poor prognosis.
Connective Tissue Diseases ; Fever ; Humans ; Meningitis, Cryptococcal ; Retrospective Studies ; Vomiting