1.Negative immune regulatory molecule TIPE2 for treating SLE mice through regulating macrophage subtype
Xingjun LI ; Yufen ZHANG ; Feng LI ; Xiaohua ZHU ; Lan HUANG
Chongqing Medicine 2017;46(24):3318-3320,3323
Objective To investigate the role of tumor necrosis factor (TNF)-alpha-induced protein 8-like 2 (TIPE2) for regulating the macrophage polarization in systemic lupus erythematosus and its curative effects on experimental SLE mice.Methods The mice were treated with activated lymphocytes derived DNA (ALD-DNA) for inducing mice model,randomly divided into AAV-scr control group and AAV-TIPE2 experimental group,and injected with AAV-TIPE2 or AAV-scr virus solution from the tail vein of mice.The expression of TIPE2 mRNA and protein in polarized macrophages,serum dsDNA antibody titer,urine protein and renal pathological index were detected.Results (1) The TIPE2 expression level of TIPE2 mRNA and protein in AAV-TIPE2-transfected cells was 13.5±1.6 times and 10.8±1.6 times of AAV-scr control group respectively.(2) M2 macrophage specific molecule MGL+ was 59.6% in AAV-TIPE2 group and MGL + cells in the AAV-scr group was 8.4%.M2/M1 odds ratio of AAV-TIPE2 experimental group to AAV-scr control group was 16.(3) The recombinant TIPE2 adenovirus related vector could stably expressed in transfected HEK-293.In vitro and in vivo experiments confirmed that AAV-TIPE2 was able to induce M2 polarization of macrophages in ALD-DNA-induced lupus mice.(4) The serum anti-dsDNA antibody,urinary protein and renal pathology in the AAV-TIPE2 group were significantly lower than those in the AAV-scr group(P<0.01).Conclusion TIPE2 alleviates the disease condition of ALD-DNA induced SLE mice through induction of macrophage polarization to M2 phenotype,which may be used as a promising therapeutic method for ALD-DNA induced SLE mice.
2.Expression of human inducible nitric oxide synthase in fibroblast cell line V_(79) and effects of H_(4)Bon iNOS activity
Feng ZHENG ; Lan HUANG ; Xiaojing WU ; Gang ZHAO
Chinese Journal of Pathophysiology 2000;0(11):-
0.05, n= 6) NO - 2 production by pcDNA 3-iNOS cells without H 4B was higher (105 58?13 33) ( n= 6, P
3.Protective effect of CpG-ODN conditioning on hypoxic/ischemicbrain damage in neonatal rats
Ying HUANG ; Lan YE ; Zhanhui FENG ; Shiping WANG ; Zhuting FENG ; Yanling JIN
Chinese Pharmacological Bulletin 2017;33(7):956-961
Aim To study the therapeutic effect of CpG-ODN, an agonist of Toll-like receptor 9 (TLR9), on hypoxic/ischemic encephapathy in neonatal rats and investigate the mechanisms.Methods Fifty healthy 7-day-old neonatal Wistar rats (in either gender, weighing 12~17g) were randomly divided into sham operation group, HIBD group, and CpG-ODN low group(0.35 mL·kg-1), CpG-ODN middle group(1.40 mL·kg-1), CpG-ODN high group(5.60 mL·kg-1).The neurological function was scored after 48h operation;ten rats of each group was executed respectively and brains tissue was taken;HE staining was used to observe the brain pathological changes.Western blot assay was used to detect the expressions of TLR9 and phosphor-p38 mitogen-activated protein kinases(p-p38 MAPK), and enzyme linked immunosorbent assay (ELISA) method was adopted to detect TNF-α expression.Results The CpG-ODN low, middle group were improved in impairment significantly compared with the HIBD group, and the brain pathological change was lessened, while the CpG-ODN high group was impaired significantly compared with the HIBD group (P<0.05), and brain pathological change was sharpened.Western blot showed the up-regulation in TLR9 and p-p38 MAPK and a significant increase of the expression of TNF-α in the brain tissue in CpG-ODN group with statistical difference in HIBD group and sham operation group(P<0.05).Conclusions The neuro-behavioral score and nervous system function can be improved and the hypoxic/ischemic brain damage can be reduced in neonatal rats in the CpG-ODN low, middle group.The protective mechanisms may be suitably via activating p38 MAPK signaling pathway to promote p38 MAPK phosphory1ation and up-regulation of the expression of TNF-α in the brain tissue of rats.
4.Clinicopathologic and immunohistochemical study of primary non-Hodgkin lymphoma of the female genital system
Xue-Feng JIANG ; Kai-Xuan YANG ; Zhi-Lan PENG ; Lian XU ; Qin HUANG ; Qian LI ;
Chinese Journal of Obstetrics and Gynecology 2001;0(04):-
Objective To investigate the clinicopathology and immunophenotype of primary non- Hodgkin lymphoma(NHL)of the female genital system,and to analyze the prognosis of such tumors. Methods Clinicopathologic features of 43 cases of primary NHL of the female genital system were studied retrospectively,with the histological classification based on the Classification of Haematopoietic and Lymphoid Tumors(WHO,2001).Immunochemistry technique,in-situ-hybridization and polymerase chain reaction methods were used to detect the immunophenotype,epstein barrvirus(EB)virus infection status and immunoglobulin heavy chain gene rearrangement,respectively.Results(1)Primary lesions:there were 24 cases of lymphoma originating in the ovary,3 cases in the endometrium,10 cases in the cervix,2 cases in the vagina and 4 cases in the vulva.(2)Staging:12 cases(28%)were in stage Ⅰ,9 cases (21%)in stage Ⅱ,and 22 cases(51%)in stage Ⅲ.(3)Histological classification:37 cases(86%)were diffuse large B cell lymphoma(DLBCL),3 cases were Burkitt lymphoma and the remaining 3 cases were unspecified peripheral T-cell lymphoma according to biopsy,immunophenotype analysis,in-situ- hybridization technique and IgH gene rearrangement detection.(4)Prognosis analysis:increase in the level of lactic acid dehydrogenase,stage Ⅲ,DLBCL and single operation suggest poor prognosis.Conclusions Establishment of the diagnosis of primary NHL of the female genital system is based on biopsy, immunophenotype analysis,in-situ-hybridization technique and IgH gene rearrangement detection,which play important roles in diagnosis and differential diagnosis of the tumor.Combined therapy is the first choice of therapeutic regimens.
5.A Review:the Molecular Mechanisms of InlA- and InlB- mediated Invasion of Listeria monocytogenes into Host Cell
Ying-Ying FENG ; Qiang ZHANG ; Lan-Hong HUANG ; Long-Juan QIN ; Qin LUO ;
Microbiology 2008;0(12):-
Gram-positive food-borne pathogen Listeria monocytogenes can invade non-phagocytic cells of the hosts by means of the special surface proteins and cause severe systemic infections. Internalins play a key role for Listeria monocytogenes in invading the non-phagocytic cells. In this study we will review and expand upon the recent advances in understanding the molecular mechanisms of InlA- and InlB- mediating the invasion of Listeria monocytogenes into host cells. This paper will also provide the theoretical base for pathogenetic mechanisms, precaution and therapy of food-borne pathogens.
6.Role of ATP-sensitive potassium channels in inhibitory effect of hydrogen sulfide on high glucose-induced inflammation mediated by necroptosis in H9c2 cardiac cells
Weijie LIANG ; Meiji CHEN ; Jianhua HE ; Wenzhu ZHANG ; Fei CHENG ; Jun LAN ; Jianqiang FENG ; Huimin HUANG
Chinese Journal of Pathophysiology 2016;32(8):1364-1369
[ ABSTRACT] AIM:To investigate the role of ATP-sensitive potassium ( KATP ) channels in the inhibitory effect of hydrogen sulfide ( H2 S) on high glucose ( HG)-induced inflammation mediated by necroptosis in H 9c2 cardiac cells. METHODS:The expression levels of receptor-interacting protein 3 ( RIP3; an indicator of necroptosis ) and cyclooxyge-nase-2 (COX-2) were determined by Western blot.The levels of interleukin-1β(IL-1β) and tumor necrosis factor-α( TNF-α) were detected by ELISA .RESULTS:After H9c2 cardiac cells were treated with 35 mmol/L glucose ( HG) for 24 h, the expression of RIP3 was significantly increased .Pre-treatment of the cells with 100 μmol/L diazoxide ( DZ; a KATP channel opener) or 400 μmol/L NaHS (a donor of H2S) for 30 min considerably blocked the up-regulation of RIP3 induced by HG.Moreover, pre-treatment of the cells with 100 μmol/L 5-hydroxydecanoic acid (5-HD; a KATP channel
blocker) attenuated the inhibitory effect of NaHS on HG-induced up-regulation of RIP3.On the other hand, co-treatment of the cells with 100μmol/L necrostatin-1 ( a specific inhibitor of necroptosis ) or pre-treatment of the cells with 100 μmol/L DZ or 400 μmol/L NaHS attenuated HG-induced inflammatory responses , evidenced by decreases in the expression of COX-2 and secretion levels of IL-1βand TNF-α.However , pre-treatment of the cells with 100 μmol/L 5-HD significantly attenuated the above anti-inflammatory effects of NaHS.CONCLUSION:KATP channels play an important role in the inhib-itory effect of H2 S on HG-induced inflammation mediated by necroptosis in H 9c2 cardiac cells.
7.Clinical application of the OPLAC for hip replacement surgery
Bo DUAN ; Sheng HUANG ; Feng LIU ; Yuanyuan OU ; Xingjin LUO ; Lan SHI
Chongqing Medicine 2015;(5):652-654
Objective To observe the efficacy ,safety and patient acceptance of the artificial airway with the oro‐pharyngo‐laryn‐gead airway cap(OPLAC) for hip replacement surgery in patients with intravenous‐inhalation combined anesthesia .Methods Sev‐enty eight cases of patients receiving the hip replacement surgery were included ,42 patients were to be adopted to establish artificial airway with the OPLAC for intravenous‐inhalation combined anesthesia ,another 36 patients treated with heath side‐lying position hypobaric spinal‐epidural anesthesia .There are two groups ,the oro‐pharyngo‐laryngead airway cap group(OPLAC ,n=42) and hy‐pobaric combined spinal‐epidural anesthesia group (CESA ,n=36) .Monitor the changes of respiratory and circulatory parameters of the two groups before the start of anesthesia ,anesthetic after 10 min ,30 min ,1 h ,and handling marrow ,observing occurrence of complications (delirium ,sore throat ,nausea ,vomiting ,deep vein thrombosis) ,following up the degree of satisfaction of patients and surgeons for anesthesia .Results The respiratory and circulatory parameters of the OPLAC group during anesthesia induction and maintenance were relatively stable ,on the other hand ,significant cyclic inhibition (blood pressure and heart rate decresing ) and re‐spiratory rate declining were observed in the CESA group after anesthesia ,cases with the use of atropine and dopamine significantly more than OPLAC group .The circulation and breath in the OPLAC group were relatively stable when handling marrow ,while cir‐culation fluctuating Significantly in the CESA group (blood pressure decreasing and heart rate increasing ) .The incidence of delirium in the CESA group was significantly higher than OPLAC group .The patient satisfaction of the OPLAC group was significantly bet‐ter than the CESA group .Conclusion The artificial airway with OPLAC for hip replacement surgery in patients with intravenous‐inhalation combined anesthesia is safe ,effective ,and satisfactory .
8.Simultaneous determination of huperzine A and huperzine B in Seventeen species from Huperzioideae subfamily by HPLC
Hui ZHAO ; Lan-Fang QIN ; Yun-Feng HUANG
Chinese Traditional Patent Medicine 2018;40(3):637-642
AIM To establish an HPLC method for the simultaneous content determination of huperzine A and huperzine B from seventeen species Huperzioideae subfamily (including 2 ineditus new species).METHODS The analysises of tartaricacid extracts of huperzine A and huperzine B were performed on a room temperature column (4.6 mm × 250 mm,5 μm),with the mobile phase comprising of methanol-spiritus mindereri flowing at 1.0 mL/min in a isocratic elution manner,and the detection wavelength was set at 310 nm.The contents were determined by external standards.RESULTS Huperzine A and huperzine B showed good linear relationships within 0.002 0-0.30 μg,0.001 8-0.27 μg (r=0.999 9),whose average recoveries were 103.86%,101.3% with the RSDs of 1.85%,1.30%,respectively.Huperzine A and huperzine B were found in seventeen species,and there were significant differences in contents from species to species.The content of huperzine A in Phlegmariurus hamiltonii was the highest,which reached up to 0.22%,with higher levels of it in Phlegmariurus cryptomerianus,Phlegmariurus petiolatus,and Phlegmariurus phlegmaria;The content of huperzine B was the highest in P.phlegmaria.CONCLUSION This accurate,stable and reproducible method can be used for the quality control of huperzine A and B from Huperzioideae subfamily.
9.BMP9 effectively induces osteogenic differentiation of immortalized calvarial mesen-chymal progenitor cells
Xudong SU ; Feng DENG ; Yinhong TANG ; Pengfei ZHOU ; Panpan LIANG ; Dan LAN ; Enyi HUANG
Military Medical Sciences 2016;40(12):984-987,993
Objective To investigate the effect of adenovirus-bone morphogenic protein 9 ( Ad-BMP9 ) on osteogenic differentiation of immortalized calvarial mesenchymal progenitor cells ( iCALs ) .Methods iCALs were infected with adenoviral vectors encoding BMP-9 or green fluorescent protein ( GFP) and the early osteogenic differentiation was assessed by detecting alkaline phosphatase (ALP) activity after being cultured for 3, 5 and 7 days.14 days after infection, alizarin red S staining was performed to study the formation of osteogenic calcium nodules .The expression of osteogenic marker genes Runx2 and OCN was assessed by quantitative real-time ( RT )-PCR and Western blotting .Results Significant increases in ALP activity and in the expressions of Runx 2 and OCN were detected in BMP-9 treated iCALs compared with GFP-treated cells(P<0.05).Meanwhile, alizarin red S staining showed that more mineralized nodules were found in the BMP-9 induced group .Conclusion BMP-9 can promote the osteogenic differentiation of iCALs .
10.The correlation study of mandibular second molar root growth and bone age in male teenagers
Ying CHEN ; Yajing ZHONG ; Shuqin LI ; Lan HUANG ; Gang FENG ; Hongwei DAI
Chongqing Medicine 2015;(28):3950-3952,3955
Objective To explore the relationship between cervical bone age and male juvenile mandibular second molar (Msm) root growth ,and to provide evidence for the development of bone and teeth in patients with orthodontic treatment . Methods The CBCT and lateral cephalometric radiographs of 114 cases of 9 .5 to 15 .5 years old children in Chongqing city and ad‐olescents cervical vertebral bone and MSM root length were measured ,using statistical analysis to explore the relationship between cervical vertebral bone ,tooth root length change of age and Msm .Results By Pearson correlation analysis ,Msm root length chan‐ges and cervical vertebral bone were positively correlated(r= 0 .737 ,P< 0 .01) .Msm root length changes showed a positive correla‐tion with age(r= 0 .681 ,P< 0 .01) .Msm root length changes was positively related with dental age(r = 0 .795 ,P< 0 .01) .Conclu‐sion The root growth of the mandibular second molar was highly correlated with cervical vertebral bone in male children .When the growth and development of patients were judged by orthodontic treatment ,we can adopt the mandibular second molar root length forecast growth level of male adolescents .