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

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

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.

Objective To study the effect of siRNA of Rab25 on apoptosis induction in ovarian carci- noma cell.Methods According to Rab25 mRNA sequence in the Genebank,the plasmids expressing siRNA against Rab25 were constructed and stably transected into A2780 cells,MTY method were applied to measure cell growth and proliferation.Apoptosis rate and cell cycle phase distribution of A2780 cells were measured by flow cytometry(FCM).Apoptosis was confirmed by agarose gel electrophoresis(AGE)of DNA.Results Cells transected with the plasmids expressing siRNA targeting Rab25 gene effectively decreased cell growth, proliferation;blocked A2780 cells in the G_1 phase of cell cycle and induced cell apoptosis.A typical DNA ladder pattern appeared on AGE.Conclusion Rab25 gene siRNA can inhibit growth and induce apoptosis in ovarian carcinoma cell line,which will facilitate further studies of Rab25 function and its application in the treatment of ovarian cancer.

Objective To discuss the initial expression of serum procalcitonin (PCT) in patients with severe traumatic brain injury (TBI) and determine the potential value of PCT to predict the neurological outcome.Methods A retrospective analysis was made on patients admitted due to severe TBI (GCS≤8 points) from July 2011 to August 2012.Mortality and neurological outcome of the survivors were determined using Glasgow outcome scale (GOS) at 6 months after TBI.Results A total of 52 patients (39 males and 13 females),at median age of 38 years (range,15-65 years) were included in the study.Twenty-eight patients had good outcome (GOS of grade Ⅳ-Ⅴ),whereas 24 patients had poor outcome or died (GOS of grade Ⅰ-Ⅲ).Within 24 hours after TBI,serum PCT level was significantly higher in patients with bad outcome compared to those with good outcome (0.778 ng/ml:0.094 ng/ml,P ＜0.01).Enhanced PCT level presented a close correlation with the poor outcome (r =0.657,P ＜0.01).Area under the receiver operating characteristic curve (ROC) was 0.879 [95％ CI (0.757,1.000)].A cutoff value of 0.2 ng/ml had a sensitivity of 100％ and a specificity of 72.2％.Once the PCT level was superior to 4.7 ng/ml,none of the patients regained consciousness.Conclusion PCT is a simple and effective method for prediction of the outcome after severe TBI.

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.

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.

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.

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.

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.