1.Inhibition of ovarian cancer growth by small interfering RNA targeting X-linked inhibitor of apoptosis gene
Jia-Jia MA ; Bi-Liang CHEN ; Xiao-Yan XIN ;
Chinese Journal of Obstetrics and Gynecology 2001;0(02):-
Objective To construct an RNA interference vector to down-regulate X-linked inhibitor of apoptosis(XIAP)gene and study the RNA interference effect on the cell cycle and growth of ovarian cancer.Methods Oligonucleotides of 64 base pairs for hairpin RNA targeting XIAP were designed, chemically synthesized,annealed,and cloned into the pSUPER vector.After identification by restriction digestion,the correct vectors were transiently transfected into SKOV3 cells,a human ovarian cancer cell line.The XIAP mRNA was detected by RT-PCR.The proteins were detected by western blot and indirect immunofluorescence staining.Flow cytometry(FCM)analysis and methyl thiazolyl tetrazolium(MTT)assay method were applied to measure cell cycle,cell growth and sensitiveness to cisplatin.Results SKOV3 cells had a high level expression of XIAP.The vector of RNA interference,which can interfere with XIAP gene was successfully constructed.After transient transfection,the expression of XIAP protein was significantly decreased in SKOV3 cells and the value of relative density was 3584?124,2138?65,1973?80 and 110 ?12,respectively(P=0.0334).At the same time,the expression of XIAP mRNA was decreased accordingly and the value of relative density was 6674?274,4532?107,2322?57 and 1864?78, respectively(P=0.0127).The FCM results showed that,the vector could increase the number of cells in G_1 phase compared with parent cells and compared with the cells transfected with pSUPER(P
2.Clinical observation of characteristic and treatment of posterior Pilon fractures.
Gao-feng YU ; Jiang-tao MA ; Min YU ; Li-qun PAN ; Liang ZHANG ; Shou-xin LIANG
China Journal of Orthopaedics and Traumatology 2015;28(6):527-530
OBJECTIVETo explore clinical characteristics and treatment of posterior Pilon fracture.
METHODSFrom January 2011 to January 2013,18 patients with posterior Pilon fracures were treated. Among them, 13 were male and 5 were female, aged from 22 to 63 years old, with an average age of 46. All the patients were closed fractures. Open reduction and internal fixation were performed after swelling subsided, lateral malleolous and posterior Pilon fracture were exposured through lateral approach on healthy side, plates were used to fixed,screws or small plates were used to fix the posterior prominence of medial malleolus after changed to supine position. AOFAS scoring were applied to evaulate clinical effects.
RESULTSAll patients were followed up with an average of 22(ranged, 12 to 48)months. All patients obtained satisfactory reset except one patient. All factures were recovered well with an average healing of 11 weeks. According to AOFAS score at the final following up, 7 cases were excellent,2 cases were moderate, and the total score was 86.8±9.2.
CONCLUSIONPosterior Pilon fracture is not rare in clinical, its mechanism of injury, traumatic anatomy, surgical procedure and prognosis are different from that of classical ankle fracture and Pilon fracture.
Adult ; Ankle Injuries ; surgery ; Female ; Fracture Fixation, Internal ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Tibial Fractures ; surgery ; Treatment Outcome ; Young Adult
3.Relation between the injected site of pertussis toxin and the induced effect on experimental allergic encephalomyelitis in rats
Taihua MA ; Jiezhong YU ; Huiii WU ; Jingmin XIN ; Liyun LIANG ; Cungen MA
Chinese Journal of Tissue Engineering Research 2006;10(14):176-178
BACKGROUND: The animals commonly used to induce experimental allergic encephalomyelitis (EAE) in oversea laboratory are rodentia animals such as Lewis rats. But in China we are short of Lewis rats. The un-susceptive animal Wistar rats are inexpensive and plentiful. The adding of pertussis toxin may induce EAE successfully in EAE un-susceptive Wistar rats.OBJECTIVE: To investigate the influence of pertussis toxin injected atdifferent sites in inducing EAE model in un-susceptive Wistar rats.DESIGN: A randomized control animal experiment.SETTING: Institute of Brain Science, Shanxi Datong University.MATERIALS: The study was performed in the Institute of Brain Science of Shanxi Datong University from March to October in 2003. Fifty-eight und adjuvant (CFA) group (n=10).METHODS: Besides routine immunization, each rat in the foot dorsum EAE group and intraperitoneal EAE group was administrated with 005 mL pertussis toxin (containing 5.0×1010 thalli), which were given intraperitoneally and subcutaneously on one hind foot respectively, and the antigen in the CFA group was replaced by CFA.cidence rate and tine of atta ck: In the foot dorsum EAE group, the incidence of EAE was 87.5% (21/24), and the time of attack was at (10.25 ±1.67) days after immunization, which were significantly different from those in the intraperitoneal EAE group [35.7% (9/24), (14.8±l.79) days, P sum EAE group, the change of body mass was (-16.00±7.30) g and the symptomscpre was 3.4±0.7, and those in the intraperitoneal EAE group Therewere no or little infiltration of inflammatory cells in the encephalon and spinal cord of CFA rats. In the EAE rats, there were inflammatory cells infiltrated in the boundary of white matter and gray matter of lumbar intumescence, spinal pia mater, spinal parenehyma, and the boundary of cerebral cortex and medulla, even deep medulla, meninges and around lateral ventricle. There were also mild inflammations in the cerebellum,brainstem and optic chiasma, which were concordant with the observed asynchronism, tic, etc. Hematoxylin and eosin (HE) staining displayed that the infiltrated mononuclear cells assembled in perivascular spaces, which were identified by morphological criteria as lymphocyte and macrophages.Forming typical muff-like changes, the inflammation was less severe in intraperitoneal EAE group than in subcutaneous foot dorsum EAE group.CONCLUSION: The EAE model induced in Wistar rats by Pertussis toxin administered subcutaneously on foot dorsum has the representative course of diseases, pathology change and clinical manifestation and the incidence of diseases is high and the cost is low. So it is a more ideal EAE model inducing method.
4.Insulin-secreting cells from induced pluripotent stem cells regulate blood glucose levels in vitro
Lei LEI ; Yingzi LIANG ; Yingjun SU ; Xianjie MA ; Xin CUI ; Shuzhong GUO
Chinese Journal of Tissue Engineering Research 2016;20(28):4210-4217
BACKGROUND:Mouse pluripotent stem cel s are induced to differentiate into insulin-secreting cel s that can effectively improve blood glucose levels in diabetic mice. OBJECTIVE:To detect mRNA and protein levels of insulin-like cel clusters from induced pluripotent stem cel s and to investigate the function of insulin-secreting cel s in vitro and in vivo. METHODS:Mouse induced pluripotent stem cel s cultured in vitro were induced to differentiate into insulin-secreting cel s using combined inducers through three stages. The morphology of endodermal cel s, islet-derived progenitor cel s and mature islet cel s in each stage was observed and relative gene expression levels were detected by PCR. Mature insulin-like cel clusters underwent dithizone staining and functions of insulin released in vitro were observed by ELISA assay. Final y, the insulin-secreting cel s were transplanted into the subrenal capsule of diabetic mice, and then blood glucose levels were observed. RESULTS AND CONCLUSION:The mature spherical insulin-like cel clusters were successful y obtained in vitro, which were in iron red by dithizone staining, and expression of insulin mRNA was determined by PCR. The insulin-like cel clusters could secrete insulin in response to various blood glucose levels by ELISA assay. In addition, after the cel s clusters were transplanted into the subrenal capsule of mice with type 1 diabetes, the blood glucose levels were marbedly improved.
5.Clinicopathologic features of intestinal dysganglionosis in children
Wuming SHEN ; Xiangru WU ; Minzhi YIN ; Jing MA ; Xin LIANG ; Chengren SHI ; Zhongde ZHANG
Journal of Clinical Pediatrics 2015;(4):312-315
ObjectiveTo investigate the clinical and pathological features of Hirschprung disease (HD), intestinal neuro-nal dysplasia (IND) and hypoganglionosis (IH) in children.MethodsThe clinical data and pathologic slices from 238 children with intestinal dysganglionosis were retrospectively analyzed. The age, sex, involved intestinal length of children and prognosis were compared.ResultsIn 238 patients, 138 (58.0%) were diagnosed by rectal mucosal biopsies. There were 122 HD patients whose median age at diagnosis was 9 months and the ratio of male to female was 4.3:1, without involvement of whole colon. There were 45 IND patients whose median age at diagnosis was 14 months and the ratio of male to female was 1.05:1, and the whole colon of 33.3% patients was involved. There were two male IH patients whose ages at diagnosis were 12 years and 18 years respectively, and their whole colon was involved. There were 59 patients with HD complicated by IND whose median age at diagnosis was 13 months and the ratio of male to female was 5.56:1 and the whole colon of 16.9% patients was involved. There were 10 male patients with HD complicated by IH whose median age at diagnosis was 11.5 months and the whole colon of 80.0% patients was involved. The ages at diagnosis, the sex ratio, the rates of whole colon involved, and the cure rates among 5 groups were signiifcantly different (allP<0.01).ConclusionsThe rectal mucosal biopsy was the main method in diagnosis of intestinal dysganglionsis in children. Patients with HD had higher incidence and mild condition and favorable prognosis. Patients with IH or patients with HD complicated by IH had lower incidence rates and severe condition and poor prognosis, followed by patients with IND or patients with HD complicated by IND.
6.Study on severe blast lung injury model of baby rabbits
Yi LIANG ; Wei DAI ; Chao MA ; Xiaojun ZHANG ; Xin YOU ; Jihong ZHOU
Journal of Regional Anatomy and Operative Surgery 2016;25(5):318-322
Objective To establish an animal model of severe blast lung injury in baby rabbits,and to provide a way to study the char-acteristic and treatment of blast lung injury in minors.Methods Randomly selected sixteen 4-weeks old New Zealand white rabbits,and the blast lung injuries were made by BST-Ⅰ biological shock tube with different drive pressure (4.0 MPa and 4.5 MPa)respectively.Then compared the injury severity of the 4.0 Mpa group and the 4.5 MPa group.Selected forty-eight 4-weeks old New Zealand white rabbits and di-vided them into the control group (8 rabbits)and the blast lung injury group (40 rabbits)Rabbits in the blast lung injury group were injured with 4.5 MPa drive pressure.Observed the vital signs,physiological index,gross anatomy of the lung,pathology,and pulmonary water content at the time of injury immediately (0 hour),2 hours,4 hours,6 hours,12 hours,24 hours,48 hours and 72 hours after the injury.Results Rabbits inthe 4.0 Mpa group and the 4.5 MPa group were all alive.The overpressure of blast wave of the 4.0Mpa group was (328.16 ± 4.78)kPa,rate of severe pulmonary defense was 12.5%,and the AIS score was (3.38 ±0.52)points.In the 4.5 MPa group,the overpressure of blast wave was (395.04 ±11.74)kPa,rate of severe pulmonary defense was 87.5%,and the AIS score was (4.13 ±0.64) points.Rabbits in the control group and the blast lung injury group were all alive.The spirits of rabbits were drooping immediately after inju-ry,and it last about 0.5 hour.Then the breathing and heart rate was accelerated,pulmonary water content was increased significantly,and there were extensive hemorrhage and edema in the lung.Most of the rabbits suffered severe lung injury,and the AIS score was (3.98 ±0.55) points.Lung tissue rupture,hemorrhage,edema,and inflammatory cells infiltration were the main pathological manifestations under light microscopy. Conclusion The model of severe blast lung injury in baby rabbits could be established with BST-Ⅰbiological shock tube and drive pressure of 4.5 MPa.It is relatively simple,easily controllable and highly repeatable,which can be used as a feasible model for the study of blast lung injury.
7.Effect of cattle encephalon glycoside and ignotin injection on brain protection in rats with traumatic brain injury
Liang TAN ; Kang MA ; Rongwei LI ; Jun TANG ; Qianwei CHEN ; Xin LIU ; Hua FENG
Chinese Journal of Trauma 2015;31(12):1118-1123
Objective To investigate the neuroprotective effect of cattle encephalon glycoside and ignotin injection on the rat model of traumatic brain injury (TBI).Methods Three hundred and six SD rats were randomly divided into sham group,TBI group,high dose group,middle dose group and low dose group according to the random number table.Rats received 1.8,0.6 and 0.2 ml/kg of cattle encephalon glycoside and ignotin injection for 28 days in high,middle and low dose groups,respectively.TBI was induced by the modified Feeney' s weight-drop method.Rat mortality,neurological function score and learning and memory ability were recorded.Brain morphological changes were evaluated with 7.0 T small animal nuclear magnetic resonance and HE staining.Evans blue was applied to assess blood brain barrier (BBB) permeability and hydrocephalus was evaluated by the brain water content.Results Mortality in high dose group decreased significantly compared to that in TBI group (22.40% vs.28.14%,P < 0.05).Defects in neurological function and learning and memory induced by TBI were significantly mitigated in middle and high dose groups (P < 0.05).Pathological damage and contralateral hippocampal atrophy in middle and high dose groups were reduced significantly compared to TBI group (P < 0.05).Hippocampus neuroapoptosis in middle and high dose groups was significantly improved compared to TBI group (P < 0.05).BBB damage was less severe in middle and high dose groups than in TBI group (P < 0.05).The treatment was preventive against secondary hydrocephalus.Conclusion Middle or high dose cattle encephalon glycoside and ignotin injection over a 28-day period has significant neuroprotective effect on the TBI rats.
8.Multivariate regression analysis of factors on long-term neurological function recovery of spinal cord cavernous hemangiomas
Yongming ZHANG ; Rongquan PEI ; Lingwen MENG ; Xin XU ; Liang MA ; Kaiwen YU ; Xiao WANG
Clinical Medicine of China 2015;(3):255-257
Objective To explore the clinical factors on the functional recovery of spinal cord cavernous hemangioma operation. Methods Fifty cases patients with complete spinal cord cavernous hemangioma were retrospectively analyzed their clinical data and its influence on prognosis of 2 years. Single factor and multiple factors analysis were performed. Results Compared with the preoperative stage,the grade of Aminoff-Logue score of 28 cases(50. 9%)had improved at least one grade,14 cases(25. 5%)patients with no significant changes and 13 cases(23. 6%)with decreased one grade at least. Logistic regression model showed that patients with clinical types(OR=3. 27,95%CI 2. 34-15. 83,P=0. 006),the time of the attacks to the surgery(OR=2. 93,95%CI 1. 41-10. 61,P=0. 012),and the attacks to the operation time( OR=4. 31,95%CI 2. 46 -12. 32,P = 0. 019 ) were related to neurological function recovery of spinal cord cavernous hemangiomas. Conclusion For patients with SCH dysfunction,regardless of what kind of clinical types of neurological dysfunction extent,should receive surgical treatment to improve neurological function recovery rate as soon as possible.
9.Analysis of 21 Cases of Risperidone-induced Leucocytopenia ADR Reports in Our Hospital
Hongyan ZHUANG ; Shanshan LIU ; Wei GUO ; Xin MA ; Hongbing HAO ; Haixia LIANG ; Qiuyan WANG
China Pharmacy 2017;28(8):1052-1055
OBJECTIVE:To analyze the features of risperidone-induced leucocytopenia ADR.METHODS:Twenty-one ADR cases of risperidone-induced leucocytopenia reported by our hospital were collected and analyzed during 2004-2015.The characteris tics of risperidone-induced leucocytopenia were discussed.RESULTS:Among 21 patients,there were 10 male and 11 female.The age was from 15 to 72 years old.Nine cases of patients were 31-40 years old (42.9%).Most of the original disease was schizophre nia.Incubation period of leucocytopenia caused by risperidone was (28.6 ± 21.4) d.Patients had no discomfort complain when leucocytopenia occurred.The lowest white blood cells reported was(3.1 ± 0.5)× 109 L-1.The leucocytopenia were improved after reduc tion,drug withdrawal and symptomatic treatment.CONCLUSIONS:Patients usually have no body discomfort complain when risperidone-induced leucocytopenia appears.Doctors should moniter ADR regularly,identify it earlier and treat carefully.
10.Roles of dCK Ser-74 in radiation-induced cell death in breast cancer cells
Nan LIANG ; Rui XIN ; Huiying XU ; Xiaodong LIU ; Bing GAO ; Wei HOU ; Xuehe ZHANC ; Shumei MA
Chinese Journal of Radiological Medicine and Protection 2012;(6):565-569
Objective To investigate the roles of dCK Ser-74 in radiation-induced cell death in breast cancer cells.Methods Different phenotypes of dCK plasmids were transfected into MCF-7 cells by liposome transfection,including dCK-Vector,dCK-WT (wild type),dCK-S74A (non-phosphorylation) and dCK-S74E (hyper-phosphorylation).All these cells were irradiated by 0,2,4,6,8 Gy X-rays,respectively.The transcriptional and translational level of dCK were detected with real time-PCR and Western blot,respectively.Radiosensitivity was analyzed using cell counting kit (CCK-8) and colony formation assays.Monodansylcadaverine staining (MDC) and flow cytometry were used to detect autophagy and apoptosis,respectively.Results Four phenotypes of dCK cell models were established successfully.After irradiation,the cell viabilities of MCF-7 and dCK-Vector decreased significantly as compared with mock group (t =14.469 and 9.357,P < 0.05),the cell viabilities of dCK-WT,dCK-S74A and dCK-S74E showed no changes (P > 0.05).The total mortalities of dCK-WT and dCK-S74E decreased significantly as compared with dCK-Vector (x2 =3.857-3.971,P < 0.05),but no changes in dCK-S74A cells (P >0.05).The apoptosis rates in dCK-S74A,dCK-Vector and control group were up-regulated after irradiation (t =-4.531,-3.688 and-7.076,P < 0.05),and the irradiation-induced apoptosis was reversed in dCK-WT and dCK-S74E (66% and 68% of the increase level in dCK-Vector group).The autophagy in dCK-WT and dCK-S74E increased by 22% and 26% (t =-9.051 and-8.411,P <0.01),but no changes were observed in dCK-S74A,dCK-Vector and control groups (P > 0.05).Conclusions The dCK-WT and dCK-S74E could reverse the irradiation-induced apoptosis,increase the autophagy occurence,and decrease the total mortality,indicating that the phosphorylation of dCK at Ser-74 sites is related to the radiosensitivity of MCF-7 cells.