To aim at the component elements in medical genetics teaching,a novel teach-ing method which introduced scientific thought was utilized.This method can promote students to assimilate the medical knowledge,know more about medical frontline information,help to raise the students’independent innovating capability,and elevate the integrative diathesis of students.

Objective To explore the clinical and molecular biological characteristics of spinocerebellar ataxia type 3(SCA3).Methods Clinical manifestation and brain MRI data of 12 patients with SCA in two families were analyged.The polymorphic CAG repeated time in the encode region of SCA3,SCA1 and SCA7 genes were compared in 15 family numbers without abnormal presentations,and 12 healthy persons of controls.Results Among 27 numbers of 4 generations in the two families had 12 patients,male and female were affected,average onset was 32 years old.The main clinic features included gait ataxia,ambiguity in speech and action clumsiness.Brain MRI showed remarkable atrophy on cerebellum and brain stem.In the two families,the CAG lengths of SCA1 and SCA7 were normal in all numbers.The repeated times of CAG of SCA3 were 11~39 in two control groups,65 ~87 in 10 cases,diagnosed as SCA3 patients.The child Ⅳ2 of family 1 was 8 years old,the repeated times of CAG of SCA3 were repeats 21 and 64 times,repectively.He might be a asymptomatic patient,because he was too young to onset the disease.Conclusions SCA3 is an autosomal dominant genetic disease.The clinical manifestations are ataxia and dysarthria.The detection of repeated times CAG can provide an effective way for the genetic and asymptomatic diagnosis.

Objective:To screen for peptides that specifically bind to PreS1 antigen from a phage-displayed peptide library.Methods: The PreS1 antigen was used as the target molecule to screen the binding peptide from the Ph.D.-7 peptide library with phage display technique,and the positive clones were identified by ELISA.Results: After three rounds of biopanning,the binding peptides were screened from the peptide library by ELISA and competitive inhibiting ELISA.Sequencing result showed that the binding peptides had high affinity and specificity.Conclusion: A peptide binding PreS1 antigen has been successfully obtained by screening the phage display library,which paves a way for the diagnosis and treatment of hepatitis B infection.

Objective: To investigate the clinical and pathological features of patients with cerebral amyloid angiopathy (CAA) related cerebral hemorrhage (C A AH). Methods : The clinical data of 5 patients with C A AH, including clinical manifestations, neuroimaging and topographical anatomy features were studied. Results: It was found that the onset and clinical manifestations of CAAH resembled hypertensive cerebral hemorrhage. CAAH could coexist with hypertension and hypertension might aggravate the pathological changes of CAA. Typical CAAH located in cortical and subcortical areas, but cerebral hemorrhage located in the basal ganglia and thalamus could not ruled out CAAH without the pathological confirmation of CAA. The neuroimages of some specific types of cerebral hemorrhage, such as insular cistern hematoma and subarachnoid hemorrhage, could manifest as hypertonial putaminal hemorrhage, but their outcomes were distinctly different. Conclusion: The causes of cerebral hemorrhage include hypertension and cerebral amyloid angiopathy, and CAA may have an important clinical value.

Objective:To study the effect of Mg 2+ on glutamate and energy metabolites during focal cerebral ischemia and reperfusion in rats. Methods: Twelve male Wistar rats were divided into 2 groups(n=6):magnesium sulfate(100 mg/kg, i.p.) group and saline group.Cerebral ischemia was produced by occlusion of middle cerebral artery with a nylon thread for 60 min and followed by 60 min reperfusion.Microdialysis probes were stereotaxically implanted into the cortex; dialysates were collected every 15 min to determine the concentrations of glucose, lactic acid and glutamate. Results: There was a dynamic decrease of glucose and an increase of lactic acid and glutamate during ischemia and reperfusion in saline group.Glucose decreased slightly in magnesium sulfate group during ischemia and recovered to normal rapidly during reperfusion. The lactic acid levels in magnesium sulfate group were lower than that in saline group during early stage of ischemia(0-15 min) and reperfusion.There were significant attenuation in the elevation of glutamate during ischemia and reperfusion when magnesium sulfate was administered and recovered to normal after 30 min of reperfusion. Conclusion: The preservation of cellular energy metabolism,the decrease of lactacidosis and attenuation of glutamate level during ischemia and reperfusion may contribute to the neuroprotective effects of Mg 2+ .

Objective To simply the method of how to obtain cultures rich in astrocytes from SD rat cerebral cortex which could be utilized in vitro experiments.Methods The neonatal rat cerebral cortex was made into suspension by mechanical dissociation,and then reduced other cells by differential velocity adherent technique,shaking in orbital shaker and passage of cultured cells.After purification,the cultured cells were identified by double immunofluorescence staining and SABC.Results We successfully obtain cultures rich in astrocytes and the proportion of astrocytes were more than 95%.Conclusion The method described above was reliable in obtaining the high purity astrocytes from neonatal rat cerebral cortex and double immunofluorescence staining was more vivid and direct.

Objective To analyse the mutation of pathogenic gene TSC2 in tuberous sclerosis complex (TSC). Methods Using polymerase chain reaction-single strand conformation polymorphism(PCR-SSCP), all the 41 exons of TSC2 gene were analyzed in 4 TSC cases(include 1 suspect case) from one family and 1 sporadic TSC case ,and compared with the kin familial controls and kinless normal controls. Results Missense mutation on exon33 1346S→P (4037T→C) of TSC2 was found in 4 familial cases, and no mutation of TSC2 gene was found in the sporadic case and all the health controls. Conclusion Missense mutation on exon33 (1346S→P,4037T→C)is a new discovery in TSC2 gene of patients with TSC.

Objective To investigate drug metabolism distribution of flurbiprofen axetil(FA) in uterine fibroids and its effect on prostaglandin E2 (PGE2).Methods A total of 86 patients with uterine fibroids from January 2014 to February 2016 in The NO.2 Hospital of Ningbo were randomly divided into control group of 44 cases and observation group of 42 cases.The control group and the observation group were given 0.9% sodium chloride solution and 1 mg/kg FA at 15 min before operation,detection of the active metabolite of FA flurbiprofen (FP),and the concentration of PGE2 in tissue homogenate was detected by ELISA.Results The observation group FP concentration of normal and tumor tissue were (0.70 ±0.13)μg/mL and (1.72 ± 0.13)μg/mL,significantly higher than the control group(0.00 ±0.00)μg/mL and (0.00 ±0.00)μg/mL, the difference was statistically significant(P<0.05), the FP concentration of tumor tissue in the observation group was significantly higher than that in the normal group, the difference was statistically significant(P<0.05), the normal tissue and tumor tissue PGE2 concentration in the observation group were (189.29 ±26.38) pg/mL and (260.01 ±46.63) pg/mL,which were significantly lower than those in the control group(210.03 ±35.22)pg/mLand(390.20 ±92.10)pg/mL, the difference was statistically significant(P<0.05), the observation group plasma FP was (5.50 ±0.72)μg/mL,significantly higher than the control group (0.00 ±0.00)μg/mL, the difference was statistically significant(P <0.05),and PGE2 was (602.38 ±84.09) pg/mL,significantly lower than the control group(920.13 ±89.05)pg/mL, the difference was statistically significant(P <0.05).Conclusion FA in the uterine fibroids have a certain distribution of targeted,can reduce the concentration of PGE2 ,so as to alleviate the pain of patients.

Objective To investigate the effect of corticotrophin-releasing factor (CRF) on the regulation of Toll-like receptor 4/NF-κB signaling pathway expression in human intestinal epithelial cell line HT-29. Methods HT-29 cells were divided into four groups, normal control group, LPS group (LPS 20 μg/ml stimulated for 24 h), CRF group (CRF 20 ng/ml stimulated 24 h) and CRF+ LPS group (CRF incubated for 12 h then changed to LPS for another 12 h). After stimulation, the expression of TLR4 mRNA of each group was examined by reverse transcriptase polymerase chain reaction (RT-PCR). Total cell protein were extracted and the expression of TLR4 and NFκB p65 at protein level were detected by western blotting.Cell culture supernatant was collected and the secretion of interleukin-8 was detected by enzyme-linked immunoasorbent assay (ELISA). Results The expression of TLR4 in LPS group at mRNA and protein level were 0.31±0.04 and 0.48±0.17,there was no significant difference compared with normal control group (0.28±0.02 and 0.45±0.12,t＝0.216 and 0.712 , P＞0.05 ). In CRF group which were 1.05±0.06 and1. 08±0.21, significantly higher than normal control group (t＝3.721 and 3.802, P＜0.05). In CRF+LPS group which were 1.68±0.05 and 1.81±0. 18,significantly higher than CRF group (t＝4. 816 and 3. 918, P＜0.05).The results of NF-κB p65 expression at protein level and interleukin-8 expression of cell culture supernatant were consistent with the results of TLR4 expression at mRNA and protein level.Conclusion CRF not only activate TLR4/NF-κB signaling pathway in human intestinal epithelial cell,but enhance the reaction of intestinal epithelial cell to LPS as well, which resulting in increased interleukin-8 secretion.

ObjectiveTo investigate the dynamic changes of T helper (Th) responses and hepatic granuloma development during Schistosoma japonicum infection and to evaluate the possible correlation between the Th1/Th2 cytokines and pathological liver damage. MethodsSera from both Schistosoma japonicum infected C57BL/6 mice and uninfected controls were measured for interferongamma (IFN-γ), interleukin (IL)-4 and IL-13 at week 4, 6, 8 and 12 post-infection. At the same time, the splenic Th1/Th2 ratio was examined. The development of liver granulomas in infected mice was also observed and the correlation between the cytokines and granulomas were assessed. Wilcoxon test and Spearman rank correlation test were performed for data analysis. ResultsSerum IFN-γ level in infected mice was (2. 835±0. 049) lgpg/mL at week 4 post infection, which was significantly higher than that in controls [(1. 787 ± 0. 174) lgpg/mL, Z= - 2. 646, P = 0. 008]. Thereafter, the level declined after week 6, but remained higher than controls at the end of 12-week experiment (Z= -2.457, P=0. 014). IL-4 and IL-13 levels did not increase until week 6 (Z=-2. 646,P=0. 008;Z=-2. 646,P=0. 008), and peaked at week 8. After soluble egg antigen (SEA) stimulation, splenic Th1/Th2 ratio in infected mice was 0.5 (95％ CI 0. 2-1.2) at week 8 and 0.3 (95％ CI 0. 3-0.6)at week 12 post infection, both of which were significantly lower than those in uninfected controls (week 8: Z=2. 173, P=0.030; week 12:Z=2.551, P=0.011). Compared with unstimulated splenic cells, splenic Th1/Th2 ratio after SEA stimulation significantly decreased at week 8 (Z=2. 236, P=0. 025) and week 12 (Z=3. 130, P=0.002). Granulomas were first observed in livers at week 6 and the size kept on increasing. A negative correlation between serum IL-13 and the diameter of liver granulomas was discovered at week 12 post infection (r=0. 636, P= 0. 048). Conclusions Th1 response is dominant during the early stage of Schistosoma japonicum infection.However,followed by oviposition, Thl response declines and meanwhile a strong Th2 response gradually develops. Therefore, Th2 response probably plays a role in the development of hepatic granulomas.