Objective To develop an approach for a rapid extraction of genomic DNA fragments from serum.Methods Serum specimen from human was processed using high temperature denaturation and was centrifuged, then using various primer sets to amplify the DNA fragment extracted, so as to verify the efficiency of this DNA extraction method.Results The templates preparing from this DNA extraction method were able to guarantee the amplification of fragments shorter than 400 base pairs.Conclusion This method is simple, rapid and inexpensive.These genomic DNA fragment extracted can be utilized to amplify definite DNA template for polymerase chain reaction.

Objective The study of loss of heterozygosity (LOH) on chromosome 1p36 was performed to locate the deletion areas probably harboring tumor suppressor genes in invasive ductal breast carcinoma not otherwise specified (IDC NOS).Methods Eighty paired breast cancer/normal tissue DNA samples were examined for LOH on chromosome lp36 using eight polymorphic microsatellite (MS) loci.The PCR products were electrophoresed on 8％ denatured polyacrylamide gel and stained using silver staining.Finally,the data were analysed and compared with the clinicopathological parameters using statistical analysis.Results In 80 IDC NOS,LOH was identified in 45 cases (56.3 ％) at least in one MS locus.MS locus D1S1310 showed the highest rate of LOH [35.7％ (25/70)].Conclusion Chromosome 1p36 might be the highly deleted region.The results of this study indicate that the chromosomal regions 1p36.23-33 might contain tumor suppressor genes associated with human breast carcinomas.

Objective To analyze computed tomography imaging features of primary adrenal schwannoma.Methods The clinical and radiological data of six cases of adrenal schwannoma confirmed by histopathology were analyzed in this study.Results All six cases of adrenal schwannoma were well-circumscribed,round-like or oval-like,solid and cystic tumors,in which two cases were with calcification,and three cases with septations.On the enhanced CT images,all six tumors displayed progressive enhancement.Conclusion Adrenal schwannoma usually presents a well-defined heterogeneous mass with cystic degeneration,sepations,calcification and characteristic progressive contrast enhancement.

Objective To investigate the roles of somatostatin(SS)positive intemeurons in the development and compensation of temporal lobe epilepsy.Methods Piloearpine-induced epilepsy rat model was established.Immunohistochemistry method was used to detect number changes and axonal sprouting of SS positive intemeurons in different domains of the hippocampus at difierent time points.Degeneration of SS positive interneurons and their neurophils were detected by the double immunofluorescence staining with SS and Fluoro-Jade B(FJB)at 7 and 60 days after status epilepticus (SE).Results In the exoerimental rat group,the number of SS positive neurons decreased in each hippocampal domain,and it reached the lowest at 7 days post-SE(There were 11.1±3.3 in hilus,2.8±0.9 in CA1region and 1.8±0.7 in CA1region,t=13.519,9.644 and 8.808,all P<0.01).In chronic phase,the number of SS neurons gradually recovered,and exceeded the control group in CA1 area at 60 days post-SE(12.8±1.5 vs 8.8±1.3,t=-4.506,P<0.01),however,the number of SS neurons in the hilus(25.5±4.6)and CA1 area(4.8±0.8)remained significantly less than normal levels(t value were 4.691 and 3.953.both P<0.01).Increased SS positive fibers were found in the lacunosum-molecular (1m)layer and outer molecular layer of dentate gyrus after 30 days post-SE,and numerous SS positive fibers were seen threnghout the layers of area CA1 at 60 days post-SE.Double immunofluuorescence revealed that a few SS positive interneurons and fibers were also labeled by FJB in area CA1 at 7 days post-SE and in CA domain/hilus at 60 days post-SE.Conclusions SS intemeurons loss plays an important role in the development of temporal lobe epilepsy.The loss is partially caIlsed by the degeneration and death of neurons;SS positive neurophils increase within area CA1 in chronic phase may play a significant role in the generation and compensation of temporal lobe epilepsy.

Objective To investigate the endoscopic and ultrasonic endoscopic features of colorectal carcinoid and the indications of endoscopic treatment. Methods The clinical data of 22 patients with colorectal carcinoid who had been admitted to our hospital from 2002 to 2007 were collected. The endoscopic and ultrasonic endoscopic features and the relationship between the features and invasion depth of colorectal carcinoid were analyzed. Results Under the endoscope, early carcinoid presented submucosa tumor with 1.5cm in diameter, and yellow or white smooth surface; advanced carcinoid presented submucosa tumor with 0.8-3.0cm in diameter, and yellow or white little nodus or ulcerative surface. The ultrasonic endoscopic feature of the colorectal carcinoid was orbicular-ovate low level echo tumor with punctiform slightly high-level echo and an unsharpness edge. Sixteen mucosal layer-cancers and submucosal layer-cancers were removed by endoscopic mucosal resection, and 10 of them were additional treated by argon plasma coagulation. After a follow-up period of 4-36 months, no recurrence was observed. Conclusions Endoscopy and endoscopic ultrasonography are effective methods to diagnose colorectal carcinoid and its invasion depth. Endoscopic treatment is a simple, safe and effective means to treat the early colorectal carcinoid tumors.

Background and purpose: Ursolic acid is widely present in spica prunellae, hedyotis diffusa and other heat antidotes. The growth of a variety of tumor cells can be inhibited and induced apoptosis by ursolic acid.This study was aimed to investigate the effect and possible mechanisms of UA on inducing apoptosis of human gastric carcinoma BGC823 cells. Methods: The MTT assay was used to detect the antiproliferative effect of UA on BGC823 cells. Flow cytometry was used to detect cell cycle and apoptosis of BGC823 cells. The expression level of bcl-2 and bax gene was investigated by real time-polymerase chain reaction (real time-PCR). Results: UA inhibited the proliferation of BGC823 cells in a dose and time-dependent way. After treatment by UA for 24. 48 and 72 h, the IC_(50) of BGC823 was 36.88, 34.72, and 32.18 μmol/L, respectively. UA could signifcantly induce apoptosis of BGC823 cells and block cells at G_2/M phase. UA could increase the expression of bax gene and decrease the expression of bcl-2 gene in a dose and time-dependent way. Conclusion: UA could induce apoptosis and inhibit the proliferation of BGC823 cells in a dose and time-dependent way. It could arrest cell cycle of BGC823 cells at G_2/M phase. Its mechanisms might be associated with the up-regulation of bax gene and down-regulation of bcl-2 gene.

AIM: G protein-coupled inwardly rectifier potassium (GIRK) channel are distributed widely in mammalian brain. In CNS, GIRK 1/2 seems to be the predominant heterotetramers which play a pivot role in the regulation of the excitability of neurons and may contribute to the resting potential by leading to a hyperpolarization of membrane potential and reduction of the action potential frequency. In the context, the Weaver mouse is the first neurological abnormality directly linked to a genetic point mutation in the GIRK2 protein which includes spontaneous seizure. GIRK2 knock out mice showed normal development but more susceptible than normal mice to seizure induced by GABA antagonist. Here, we report that the mRNA and protein expression of GIRK subunit 2 is altered in kainic acid(KA)-induced epileptic rat hippocampus. METHODS: Rats were injected with kainate 14 mg/kg intraperitoneally to establish an acute and chronic temporal lobe epilepsy model. At chronic spontaneous seizure stage, by using of in situ hybridization, immunocytochemistry and Western blotting, the GIRK 1,2 mRNA and protein were analyzed quantitatively in the dentate gyreus, CA1, CA3 regions of hippocampus. RESULTS: GIRK1,2 mRNA and proteins were expressed abundantly in all regions of hippocampus. KA induced seizures and caused a significant increase in GIRK2 mRNA abundance and immunoreacitivity; only GIRK1 mRNA was increased significantly, but no difference was found by Western blotting protocol. CONCLUSION: GIRK1,2 mRNA and protein expression in the hippocampus of epileptic rat brain is up-regulated, which may be an adaptive response to over-excitability of neuron networks and prevent the over-excitability spread in hippocampus (DG-CA3-CA1). [

Objective To explore the treatment using two different ventilation tubes insertion for secretory o-titis media ,and to determine which factors are associated with the tympanic membrane perforation after a extrusion or surgical removal of a ventilation tube(VT) .Methods A retrospective analysis of 121 cases (210 ears) with re-fractory secretory otitis media from 1998 June to 2013 March was carried out .There were 58 cases (105 ears ) and 63 cases (105 ears) in patients and control group ,respectively .All subjects had more than 2 years of history ,and were treated by two types of ventilation tube (Goode -T and Shepard tube) .Various factors were analyzed to deter-mine the factors associated with persistence of a tympanic membrane perforation after VT extrusion or removal ,in-cluding gender ,age ,medical history ,ventilation tube pattern ,location ,history of previous VT insertions ,intuba-tion period reason for VT insertion ,condition of the TM ,nature of the tympanic cavity effusion and previous ade-noidectomy ,et .Results Analysis factors of perforation group (105 ears) and the control group (without perfora-tion ,105 ears) ,were statistically significant in ventilation tube pattern (χ2 =6 .916 ,P<0 .01) ,history of previous VT insertions(χ2 =7 .325 ,P<0 .01) ,frequency of previous VT insertions (χ2 =13 .01 ,P<0 .01)and the nature of the tympanic cavity effusion(χ2 =7 .04 ,P<0 .01) .The proportion of perforation group about Goode - T ventilation tube ,history of previous VT insertions ,the frequency of tube (more than 3 times) ,mucus of the middle ear cavity were 58 .4% ,57 .3% ,66 .7% and 57 .9% respectively ,others had no significant difference between the two groups . The multivariate analysis showed that ,regardless of the ventilation tube ,the frequency of tube was the tympanic membrane perforation risk factor ;Goode-T tube increased the risk of perforation of tympanic membrane was 1 .637 times than Shepard tube(OR=1 .637 ,P<0 .01) .Conclusion Using the Goode - T type ventilation tube ,history of previous VT insertions ,the frequency of tube (more than 3 times) ,mucus of the middle ear cavity were four risk factors for persistence of a tympanic membrane perforation after VT insertion for secretory otitis media .

Objective To explore the clinical features and outcomes of patients with cerebral venous sinus thrombosis (CVT) during pregnancy and puerperium.Methods A retrospective study was performed in 24 cases of pregnant women with CVT among 15 625 deliveries in Xuanwu Hospital fromJanuary 2002 to October 2009,including 7 cases happened during pregnancy and 17 during puerperium.The etiology,clinical presentations,imaging examination results,other relevant examinations,and pregnant outcomes of these patients were analyzed.Results (1) Incidence and etiology:the incidence of CVT during pregnancy and puerperium was 0.15% (24/15 652) and 29% (7/24) of the patients fell ill during pregnancy and 71% (17/24) during puerperium.Five were complicated with severe preeclampsia,while another 5 complicated with hyperemesis.One woman was complicated with anemia.CVT was identified after spontaneous delivery in 9 cases and 8 after cesarean section.(2) Clinical presentations:Among the 24 CVT cases,22(92% ) suffered from headache,16(67% ) reported nausea and vomiting,15(63% )experienced hyperspasmia and 6 (25%) complained of blurred vision.On admission,8 (33%) patients were unconscious,3 (13%) with hemiplegia,and 8 (33%) were febrile.(3) Imaging and laboratory examinations:twelve patients underwent digital subtraction arteriography ( DSA) and were diagnosed.Fifteen women showed superior sagittal sinus thrombosis in MRI and magnetic resonance intravenous angiograph(MRV).Examination of the fundus found papilledema in 4 cases.Normal cerebral sinus fluid and laboratory routine tests were reported in 13 cases,but 6 cases of hyperlipidemia,7 cases of abnormal activated partial thromboplastin time ( APTT),5 cases of abnormal international normalized ratio (INR),3 cases elevated platelet count,4 cases of positive D-dipolymer,1 cases of low hemoglobulin level ( <10 g/L),4 cases with abnormal hematocrit,and 10 cases of elevated fibrinogen ( > 4 g/L) were identified.(4) Management and pregnancy outcomes; among the 7 cases happened during pregnancy,2 were complicated with severe preeclampsia and delivered through cesarean section immediately and discharged after proper management including depressurization,spasmolysis,dehydration and anticoagulant therapy.Five of the 7 cases presented with CVT during early pregnancy,among which 1 was discharged after dilation and curettage followed by anticoagulant therapy,2 received endovascular thrombolysis after which one was discharged and the other one left with right hemiparesis and 2 patients died.Among the 17 patients presented with CVT during puerperium,10 received anticoagulant therapy after which 4 were fully recovered,5 left with functional disturbance (3 with hemiplegia,1 with incomplete motor aphasia and hemiparesis and 1 with blurred vision) and one died.Among the rest 7 cases who underwent endovascular thrombolysis,3 were fully recovered,3 left with functional impairment ( 1 with blind and headache and 2 with hemiplegia),and one died.Altogether,there were 14 patients underwent systemic anticoagulant therapy after which 7 were fully recovered,5 left with dysfunction and 2 died.Among the 10 cases received endovascular thrombolysis,4 were fully recovered,4 remained some dysfunction and 2 died.Conclusions CVT,mostly presented as superior sagittal sinus thrombosis,are more common in puerperium than during pregnancy and DSA is the golden standard for the diagnosis of CVT.Anticoagulant therapy and endovascular thrombolysis are effective in the treatment of CVT,but may left the patients with functional disturbance or even death.Prompt diagnosis and treatment ensure a better outcome for pregnant women complicated with CVT.

Objective:Based on the survey of qualitative and quantitative on rural doctors in 6 provinces and cities in China , this paper aimed to compare the present situation of construction of rural doctors under the new health care reform and explore the impact of the new health care reform as well as to propose some suggestions . Methods:Based on the survey of questionnaire and in -depth interviews on rural doctors in 6 provinces and cities in China .Results:The problems existed for rural doctors in China such as insufficient number of personnel , ag-ing, lack of young rural doctors supplement , low level the requirements and situation of professionalism , etc.Con-clusion:It should accelerate the process of the cultivation of rural doctors , promote the education level of rural doc-tors, gradually ease the current situation of the rural doctor shortage .