1.Construction of Thymosin ?4 Shrna Lentivirus and Its Effects on the Expression of Thymosin ?4 in 293T Cells
Sijian HUANG ; Chang LIU ; Zelian QIN
Chinese Journal of Minimally Invasive Surgery 2005;0(07):-
Objective To construct a recombinant lentiviral vector for human thymosin ?4 (TMSB4) and to test the mRNA and protein expression of TMSB4 in 293T cells after being infected by shRNA lentivirus. Methods We designed a specific sequence of small hair RNA targeting TMSB4 gene; the complementary DNA containing both sense and antisense oligo DNA of the targeting sequence was cloned into the pGCSIL-GFP vector to construct a lentiviral vector. The vector was converted into the competent DH5a coli,which confirmed by PCR and sequencing. Then the viral vector and the packed systemic vector were cotransfected 293T cells to get the lentivirus. The virus titer was determined. Afterwards,in the 293T cells infected with the lentivirus,the expression of TMSB4 was detected by real time PCR and immunocytochemistry. Similarly,a primary fibroblast was also infected with the lentivirus. Results Compared with negative control cells,the mRNA and protein levels of TMSB4 in 293T cells infected with the lentivirus were reduced by 44% (0.56?0.11 vs 1.00?0.06,F=89.673,P
2.Factors Associated with Postvitrectomy Endophthalmitis
Sijian HUANG ; Sufen LU ; Mingse LIN ; Suhong WU
Chinese Journal of Nosocomiology 1994;0(04):-
OBJECTIVE To investigate factors associated with postvitrectomy endophthalmitis. METHODS Cases with postvitrectomy endophthalmitis in our hospital between 2003 and 2007 were collected and retrospectively reviewed. RESULTS All 14011 cases (eyes) underwent pars plana vitrectomy and postoperative endophthalmitis happened to seven cases (eyes). The incidence of infection was 0.047%. Confirmed microbiology was demonstrated in 4 of 7 cases (57.0%),in which Gram-positive cocci were found 75.0%. 71.4% Of cases with postvitrectomy endophthalmitis were older than 60. The infection was controlled in 6 cases after antibiotic treatment. One case with diabetes and culture confirmation of Gram-negative bacilli deteriorated even after the second vitrectomy,and evisceration was finally performed. CONCLUSIONS The predominant organism is Gram-positive cocci in postvitrectomy endophthalmitis. The risk factors include increasing age,presence of systemic diseases and immunodeficiency. Timely and effective antibiotic therapy is important to restore visual acuity for postvitrectomy endophthalmitis.
3.Correlation research between CTLA-4Ig impacting on Th17 and Treg cells expressed in spleen of lupus mice and its intervention role in lupus-like symptoms of B6.MRL-Faslpr/J lupus mice
Fang WANG ; Cuili HUANG ; Youkun LIN ; Yanyan LUO ; Sijian WEN ; Hui LIU ; Xiaogeng HUANG
Chinese Journal of Immunology 2015;(1):40-44
Objective:To observe the correlation between the B7 blocker,CD28 and cytotoxic T lymphocyte-associated antigen 4 immunoglobulin (CTLA-4Ig) impacting on Th17 and Treg cells expressed in the spleen of lupus mice and its intervention role in lupus-like symptoms of B6.MRL-Faslpr/J lupus mice.Methods:Sixteen 4-month-old female B6.MRL-Faslpr/J mice were selected and randomly divided into treatment group ( groupⅠ) and control group ( groupⅡ);injected the same amount of CTLA-4Ig and PBS intra-venously,checked their 24 hour urine protein ,ANA antibody,ds-DNA antibodies before and after the intervention.Two weeks after the intervention ,detected serum IL-17A,and the percentage of Th17 and Treg cells in their spleen.Results:Two weeks after the last inter-vention,24-hour urine protein,serum ANA and ds-DNA in groupⅠdecreased,and all the differences were statistically significant (P<0.05) compared with groupⅡ.Two weeks after the last intervention,serum IL-17A and the proportion of Th17 cells in the spleen in groupⅠwere lower than those in groupⅡ, but Treg cells in CD4+T lymphocytes was higher than that in groupⅡ,all the differences were statistically significant ( P<0.05).Conclusion:CTLA4-Ig can relieve the lupus-like symptoms in lupus mice;raising the number of Treg cells and decreasing the number of Th17 cells may be one of the important mechanisms for CTLA-4Ig to alleviate lupus-like symptoms in B6.MRL-Faslpr/J mice.
4.Effects of arsenic trioxide on the mRNA expression of toll like receptor-9 and interferon regulatory factor-5 in patients with systemic lupus erythematosus
Sijian WEN ; Wenjun ZHENG ; Youkun LIN ; Meiqin XU ; Cuili HUANG ; Hui LIU
Chinese Journal of Rheumatology 2014;18(4):227-231,后插1
Objective To investigate the mRNA expression of toll like receptor-9 (TLR9) and interferon regulatory factors-5 (IRF5) of AS2O3 on peripheral blood mononuclear cells (PBMCs) in systemic lupus erythematosus (SLE) patients.Methods PBMCs of 15 SLE patients and 15 healthy subjects were treated with different concentrations of AS2O3 and cyclophosphamide (CTX) in vitro.Real-time quantitative polymerase chain reaction was used to amplify TLR9 and IRF5 gene before and after 12 and 24 hours drug intervention and the mRNA expressions were measured.Differences between groups were analyzed by paired t test or variance analysis.Results The mRNA expression levels of TLR9 [12 h(1.38±0.26) and 24 h (1.28±0.35)] on PBMCs in SLE patients were significantly higher than those in healthy controls [12 h(1.05±0.35) and 24 h (0.97±0.19)](t=2.37,P=0.03; t=2.44,P=0.02).The IRF5 mRNA expression levels [12 h (0.95±0.27) and 24 h (0.91 ±0.35)] in SLE patients were obviously higher than those in healthy controls [12 h (0.62 ±0.23) and 24 h (0.60±0.39)] (t =3.07,P=0.01 ; t =3.45,P<0.01).AS2O3 could suppress the mRNA expression of TLR9 on PBMCs and the effect was gradually increasing with the increasing concentration of AS2O3 and processing time [0.2 mg/L AS2O3 group 12 h (0.430±0.110) and 24 h(0.290±0.050),0.4 mg/L AS2O3 group 12 h (0.170±0.038) and 24 h (0.090±0.017),0.8 mg/L AS2O3 group 12 h (0.023±0.011) and 24 h (0.003±0.001)].Comparing with CTX [12 h (0.814±0.081) and 24 h(0.755±0.139)],AS2O3 had a more significant strong effect on inhibiting the expression of TLR9 mRNA in SLE patients [F=165.32(12 h),P<0.01; F=99.20 (24 h),P<0.01].The mRNA expression of IRF5 on PBMCs was not suppressed by AS2O3 and CTX and there was no statistically significant difference between groups (P>0.05).Conclusion There is abnormal expression of IRF5 and TLR9 mRNA in SLE patients.AS2O3 may suppress the TLR9 mRNA expression in SLE patients,which may be one mechanism of clinical effectiveness.
5.Clinical features and treatment of 12 cases with incontinentia pigmenti-associated retinopathy
Qiong WANG ; Xiaoyan DING ; Sijian HUANG ; Xiaoyu LI ; Limei SUN ; Songshan LI ; Ting ZHANG ; Li HUANG ; Jiaqing LI
Chinese Journal of Ocular Fundus Diseases 2021;37(4):277-283
Objective:To observe and analyze the clinical features, treatment methods and efficacy of patients with retinopathy associated with incontinentia pigmenti (IP).Methods:A retrospective case study. Twelve clinical confirmed IP patients (24 eyes) in Zhongshan Ophthalmic Center of Sun Yat-sen University from January 2015 to December 2018 were included in this study. The best corrected visual acuity and intraocular pressure examination were performed in patients (>4 years old). All patients were examined on the anterior segment, vitreous body, and fundus under topical anesthesia or general anesthesia. Eight cases underwent genetic testing. Patients with active disease should be given anti-vascular endothelial growth factor (VEGF) drug treatment, retinal laser photocoagulation or vitrectomy, those without active disease should be observed. All patients were followed up for 1 to 3 months, with an average follow-up time of 18.7 months.Results:All patients were all female, with an average age of 6.3±9.8 years old at the first ophthalmology visit. According to the recommendations of the pediatrician, 3 cases were actively screened for ophthalmology (referrals), with an average age of 0.4±0.5 years (median age: 2 months). A total of 9 cases were not recommended for referrals (non-referrals), including 3 cases of ophthalmology who were diagnosed for the first time due to visual impairment, and 6 cases of undiagnosed IP before the ophthalmology visit, the average age of their first visit was 8.2±10.8 years (medium age: 3 years old). The age of the first visit for non-referred patients was larger than that of referrals, and the difference was statistically significant ( Z=-2.141, P=0.036). Among the 24 eyes of 12 cases, there were no obvious fundus abnormalities in 1 case or 2 eyes, 11 cases of IP-related retinopathy in 22 eyes (91.7%, 22/24), 8 cases of binocular asymmetry (66.7%, 8/12). There were active lesions on the fundus in 7 eyes (29.2%, 7/24). Patients underwent simple retinal laser photocoagulation and/or anti-VEGF drug therapy. During the follow-up, retinal neovascularization recurred in 1 eye. Among the 8 cases that underwent genetic testing, 3 cases (37.5%, 3/8) were deleted in exons 4-10 of the IKBKG gene. Conclusions:IP is more common in women. IP-associated retinopathy is noted with early-onset, asymmetrical retinopathy, which is identified with retinal neovascularization and vitreous proliferation. Early detection and timely treatment are essential.
6.Protective effect of Acronychia pedunculata water extract on photoaging keratinocyte model and its mechanism
Xiuling LUO ; Yuan ZHOU ; Binbin ZHAO ; Manli HUANG ; Wenyu LI ; Runge FAN ; Sijian WEN ; Jichao LI ; Huayu WU ; Youkun LIN
Chinese Journal of Medical Aesthetics and Cosmetology 2022;28(6):517-521
Objective:To investigate the protective effect and mechanism of Acronychia pedunculata water extracts on UV-induced light damage of human keratinocytes.Methods:The experiment was conducted from December 2018 to April 2020 in the Guangxi Medical University Laboratory of Genetics. The photoaged keratinocyte model was used, the cells were co-cultured with different concentrations of Acronychia pedunculata water extracts. The cell proliferation rate was detected by CCK-8 method. The levels of reactive oxygen species (ROS), malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX) and total antioxidant capacity (T-AOC) of cells were detected by a test kit. The levels of IL-1β, IL-6 and tumor necrosis factor-alpha (TNF-α) were determined by ELISA.Results:The proliferation of HaCaT cells was promoted by 0.5 mg/L-2.0 mg/L of the extracts. Compared with control group, the proliferation rate of HaCaT cells in the experimental group was significantly increased ( P<0.05). Compared with control group, the contents of ROS was decreased ( F=214.67, P<0.05), MDA was decreased ( F=811.88, P<0.05), SOD was increased ( F=28.95, P<0.05), CAT was increased ( F=213.31, P<0.05), GPX was increased ( F=65.10, P<0.05), T-AOC was increased ( F=305.58, P<0.05), IL-1β was decreased ( F=15.46, P<0.05), IL-6 was decreased ( F=59.2, P<0.05), and TNF-α was decreased ( F=33.13, P<0.05). Conclusions:The extracts of 0.5-2.0 mg/L of Acronychia pedunculata have protective effects on the photoaging cell model, which may be related to the increase of SOD, CAT, GPX and other antioxidant enzymes and the level of T-AOC in photoaging HaCaT cells, and the decrease of ROS, MDA content and the expression of inflammatory cytokines.
7.Complications of deep brain stimulation internal pulse generator replacement procedures
Yunxuan CAI ; Dianyou LI ; Shikun ZHAN ; Sijian PAN ; Wei LIU ; Peng HUANG ; Xiaoxiao ZHANG ; Bomin SUN ; Yixin PAN
Chinese Journal of Neuromedicine 2018;17(10):1024-1027
Objective To investigate the complications of deep brain stimulation (DBS) internal pulse generator (IPG) replacement procedures and discuss the reasons,preventive measures and treatments.Methods From 2012 to 2016,285 procedures (according to the number of replacement IPG) were performed for 211 patients in our hospital.Among them,178 patients were with Parkinson's disease,29 patients were with dystonia,3 patients were with tic disorder,and one with essential tremor.Thirty-two patients previously used Medtronic replaced with local DBS with brand of PINC and Sceneray,and the other 179 patients remained the use of Medtronic brand.Furthermore,36 patients got extension cable reimplantation along with IPG replacements.Results Replacement surgeries were divided into 3 types:IPG replacement in situ;bilateral side single-channel IPG was replaced by double-channel IPG or double-channel IPG was replaced by bilateral single-channel IPG;extended cables and IPG replacement were carried out simultaneously.The follow up period was from one to 6 years for these 211 patients,and 15 got surgical-and hardware-related complications:6 with sack hemorrhage,2 with skin erosion,one with IPG rejection,3 with adaptor fracture,and 3 with impedance abnormality were recorded;no infection was noted.Twenty-six patients got significant improvement after new IPG replacement.Conclusions DBS IPG replacement operations is a regular surgery with certain safety.Personalized surgical procedures,rigorous intraoperative operation and correct postoperative management can effectively reduce and prevent the complications of IPG replacement surgery.
8.Meta-analysis of the Efficacy of Gongliuqing Capsules Combined with Mifepristone in the Treatment of Uterine Leiomyoma
Yizheng ZHONG ; Jiahua HUANG ; Meijun PAN ; Huaqing JIANG ; Hao ZHENG ; Wei LI ; Sijian CHEN ; Yingying CAO ; Fan HUANG ; Fasen DENG ; Weipeng SUN ; Kaimin GUO
China Pharmacy 2020;31(2):221-226
OBJECTIVE:To systematically evaluate therapeutic efficacy of Gongliuqing capsules combined with mifepristone in the treatment of uterine leiomyoma ,in order to provide evidence-based reference for clinical medication. METHODS :Retrieved from Cochrane Library ,PubMed,Embase,CJFD,VIP,CBM and Wanfang database ,randomized controlled trials (RCTs)about Gongliuqing capsules combined with mifepristone (trial group )versus mifepristone alone (control group )in the treatment of uterine leiomyoma were collected. After literature screening and data extraction ,the quality of included literatures was evaluated with modified Jadad scale. Meta-analysis was conducted by using Stata 14.0 software,and trial sequential analysis (TSA)was performed by using TSA 0.9 software. RESULTS :A total of 12 RCTs were included ,involving 1 210 patients. The results of Meta- analysis showed that the total response rate of trial group [RR =1.12,95%CI(1.00,1.26),P<0.05] was significantly higher than that of control group ;maximum uterine leiomyoma volume after treatment [SMD =-1.08,95%CI(-1.21,-0.95),P<0.05],uterine volume after treatment [SMD =-0.80,95%CI(-1.14,-0.45), P<0.05],follicle stimulating hormone (FSH)level [SMD = - 0.28,95% CI(- 0.45,- 0.19),P<0.05],luteinizing hormone(LH)level [SMD =-0.44,95%CI(-0.52,-0.12), 020-38076311。E-mail:867203217@qq.com P<0.05],E2 level [SMD =-2.69,95%CI(-3.08,-1.49), P<0.05] and progesterone (P)level [SMD =-1.27,95%CI(-1.69,-0.71),P<0.05] of trial group were significantly lower or better than those of control group. Results of subgroup analysis showed that except for the level of FSH in 5 and 10 mg mifepristone groups (P>0.05),maximum uterine leiomyoma volume after treatment ,uterine volume after treatment ,the levels of FSH,LH,E2 and P in trial group were significantly lower than control group. The results of TSA showed that there were definite evidences for total response rate of Gongliuqing capsules combined with mifepristone being better in the treatment of hysteromyoma. CONCLUSIONS :Total response rate of Gongliuqing capsules combined with mifepristone in the treatment of hysteromyoma is better than mifepristone alone ,which can effectively decrease the volume of maximum uterine leiomyoma volume and uterine vilume ,and reduce the level of serum hormone.
9.Comparison of clinical efficacy between HLA-mismatched related and HLA-matched unrelated donor hematopoietic stem cell transplantation for hematopoietic malignancies.
Sijian YU ; Min DAI ; Jing SUN ; Zhiping FAN ; Fen HUANG ; Yu ZHANG ; Qianli JIANG ; Hongsheng ZHOU ; Dan XU ; Fanyi MENG ; Qifa LIU
Chinese Journal of Hematology 2014;35(9):826-830
OBJECTIVETo compare the clinical efficacy of HLA- mismatched related donor (MRD) and HLA-matched unrelated donor (MUD) hematopoietic stem cell transplantation (HSCT) for hematopoietic malignancies.
METHODS174 patients with hematopoietic malignancies undergoing allogeneic HSCT (allo-HSCT) (82 from MRD and 92 from MUD) between June 2002 and December 2012 were enrolled in this retrospective study. Hematopoietic engraftment, graft versus host disease (GVHD), relapse, overall survival (OS) and disease-free survival (DFS) were compared between MRD and MUD group.
RESULTSThere was no significant difference between MRD and MUD group in terms of age, gender, disease type and disease status before transplantation (all P>0.05). The incidence of Ⅰ-IV acute GVHD (aGVHD) was 62.2% and 54.3% in MRD and MUD group (P=0.295); the incidence of III-IV aGVHD between the two groups was 15.9% and 9.8% (P=0.229). The incidence of chronic GVHD (cGVHD) was 28.4% and 45.1% in MRD and MUD group (P=0.036), but there was no significant difference in the incidence of extensive cGVHD between the two groups (9.0% vs 12.2%, P=0.525). The mortality of GVHD was 8.5% and 10.9% in MRD and MUD group (P=0.605). The 10-year OS and DFS were (50.1±6.1)% and (48.8±6.1)% in MRD group, compared with (50.5±6.7)% and (46.3±6.2)% in MUD group (P=0.501, P=0.873, respectively). The 10-year cumulative relapse rate was (21.5±5.7)% and (37.6±7.3)% in MRD and MUD group (P=0.194).
CONCLUSIONMRD is equivalent to MUD in efficacy and safety. Without HLA- matched related donors, MRD is superior to MUD because donor source is unlimited and transplantation could be made promptly according to disease status.
Adolescent ; Disease-Free Survival ; Graft vs Host Disease ; Hematologic Neoplasms ; therapy ; Hematopoietic Stem Cell Transplantation ; Histocompatibility Antigens Class I ; immunology ; Humans ; Neoplasm Recurrence, Local ; Retrospective Studies ; Transplantation, Homologous ; Treatment Outcome ; Unrelated Donors