Objective To study the effect of of human cytomegalovirus (HCMV) infection on the expressions of Hox genes.Methods Forty eight kunming mice were randomly divided into infection group (n=32) injected with HCMVAD169 and control group (n=16) injected with saline into their brain.After 7,15,30,and 60 days,the cerebral lesions were observed by pathological method.HCMV antigen was detected by immunohistochemical method and HCMV DNA was detected by polymerase chain reaction (PCR). On the basis of developing HCMV mouse model. reverse trancriptase-polymerase chain reaction (RT-PCR) was applied to determine the expressions of Hox gene in the brains of infected mice.The expression of Hox genes were also analysed with Northern-blot by isotope labelled Hox genes oligonucleotide probes. Results A HCMV infection model was developed and extensive pathological damages in brain tissue of infected mice were observed.Meanwhile.the HCMV-LA and HCMV-DNA were also found in brain tissues of HCMV infected mice.The expression level of Hox genes in control and infected mouse brain were determined by RT-PCR and Northern-blot.RT-PCR and Northern-blot showed that mouse brain expressed Hox-A9,Hox-A10,Hox-A11,Hox-A12,and Hox-A13,but they did not express Hox-B13.After HCMV infection,murine brain was induced to express Hox-B13 gene(P＜0.01),and reached the peak at 30 d after infection.Comparing with the control group,the expression of Hox-A9 and Hox-A11 were down-regulated in infected group (P＜0.05);the expression of Hox-A10 and Hox-A13 were significantly higher in infected groups (P＜0.05).Conclusion The results suggest that HCMVAD169 is able to cause mouse CNS infection and induced the abnormal expressions of Hox genes. which provides more information for understanding the mechanism of congenital abnormal due to HCMV infection and a valuable method of clinical prevention and treatment of HCMV infection.

Object To develop a best approach for the rapid propagation of Lilium brownii F.E. Brown var. viridulum Baker by tissue culture. Methods Different parts of the bulb were tried as the explant and cultured on different culture media with the additon of different portions of various hormones at various cultural conditions. Results The best medium for the culture of explant bulb was MS+NAA 0.5 mg/L and GA 3 2.0 mg/L and the highest induction rate was at the lowest part of the scale leaves, which attained 92.5%. The small bud can further differentiate to form secondary small buds. By liquid-quivering culturing, the weight increase can be accelerated. Conclusion Tissue culture of L. brownii var. viridulum can achieve its rapid propagation, resulting in the possibility for its industrial production.

Objective To determine the efficacy and safety of Chinese Medicine Jiaweidingchuan Soup in the treatment of asthma.MethodsPatients(n =32)with mild-to-moderate asthma were treated with Chinese Medicine Jiaweidingchuan Soup for 4 weeks.We measured the FVC、FEV1、PEF、IL-6 and IFN-? level.ResultsAfter treatment,the FVC、FEV1 and PEF values and the expression of IFN-? in patient with asthma increased significantly(P

Objective To explore the expression of the CENP-W in gliomas and investigate the effects of its invasion. Methods The expression level of the CENP-W in gliomas with varied pathologic grade were detected by immunohistochemical analysis,RealTime PCR,and Western Blotting. U251 cells were transfected with the specific siRNA to repress the CENP-W expression level. The invasion ability of U251 cells were examined by Transwell Chamber assay ,while RAS mRNA and protein levels were detected at the same time. Results The expression levels of the CENP-W in glioma tissues were significantly high and the CENP-W gene could enhance the invasion of U251 cells . The expression of RAS was down-regulated when the expression of CENP-W was repressed. Conclusion The CENP-W has an oncogenic role in human brain gliomas and may regulate the invasion of gliomas by adjusting the RAS signaling pathways.

Objective To evaluate the safety and effectiveness of three endoscopic methods for re-moval of common bile duct stones (CBDs)accompanied with periampullary diverticula(PAD).Methods A total of 154 patients hospitalized at Taizhou Hospital and Taizhou No.1 People′s Hospital of Zhejiang prov-ince from December 2012 to July 2013 were divided randomly into three groups,i.e.,EST,EPBD and ES-BD group,and received the treatment of EST,EPBD and limited EST plus EPBD (ESBD)to extract CBDs, respectively.After 12 months of follow-up,the rate of full stone clearance,stones clearance rate in one time,the rate of mechanical lithotripsy,the rate of urgent lithotripsy,the average procedures,the average removal time and the complication incidence among three groups were compared.Results The rates of stone clearance in one time in group ESBD was higher than those of group EST and group EPBD (94.12% VS 78.43%,73.08%;P <0.05)with significant difference.The average procedures in group ESBD was lower than that of group EPBD (1.08 VS 1.31,P <0.05),which also showed significant difference.The occur-rence rates of early complication in group ESBD was lower than that of group EPBD (15.69% VS 34.61%, P <0.05).The occurrence rates of post-ERCP hyperamylasemia in group ESBD was lower than that of group EPBD (5.88% VS 21.15%)with significant difference (P <0.05).The incidence of pneumobilia in group EST was higher than those of group EPBD (52.27% VS 26.19%,P =0.013)and group ESBD (52.27%VS 27.66%,P =0.016).Conclusion The stone extraction efficiency of ESBD is better than that of EST and EPBD.Compared with conventional EST,ESBD shows similar safety level,and is safer than EPBD.So ESBD is a safe and effective method to remove CBDs with PAD.

Objective:To investigate the effects of iron deficiency in early life on neurobehavioral manifestations in CDH2 genetic mutation rats, in order to explore interaction between iron deficiency and CDH2 gene mutation in autism-like behavior.Methods:The 16 female SD rats with established CDH2 genetic mutations were randomly divided into 8 rats as iron deficiency group and 8 rats as normal diet control group.After mating, rats were fed low-iron diets during pregnancy and lactation or standard diets as control.The offsprings were randomly divided into 27 rats as iron deficiency group and 27 rats as control group.The voice communication ability of offspring was studied through ultrasonic vocalizations.The data and videos of open-field test, three-chamber social interaction test were recorded by animal behavioral video analysis system(Smart3.0). The characteristics of behavior changes were analyzed by statistical methods.Results:Total number of calls in rats aged 10 day in iron deficiency group[(755.67±161.86)times]were significantly lower than that in the control group[(1461.89±166.57)times]( P<0.05). Total calling duration[(41.77±16.17)s]were significantly lower than that in the control group[(86.22±10.07)s]( P<0.05). There is a certain synergistic effect of CDH2 genetic mutation and iron deficiency on the reduction of total calls in rats aged 6 and 8 day( P<0.05). The total distance, distance in zone-periphery, mean speed in zone-total and mean speed in zone-periphery of rats in iron deficiency group were all higher than that in the control group( P<0.05); the numbers of standing and rearing were reduced( P<0.05). In the second stage of the three-chamber test, the rats in the iron deficiency group had shorter communication time with the new stranger 2 and longer interaction time with the old stranger 1 compared with the control, showing a weakening trend of social ability, but the difference was not statistically significant( P>0.05). Conclusion:Autism-like behavioral changes occurred in rats with iron deficiency in early life and CDH2 genetic mutation, and there is a certain degree of synergy.Iron deficiency and CDH2 mutation may increase the risk of autism spectrum disorder.

Objective:To evaluate the effect of combined use of Guben-Tiaoping Decoction on the success rate of stepwise treatment and life quality of asthma patients. Methods:A total of 76 patients with asthma in remission stage who were using ICS were divided into treatment group (38 cases) and control group(38 cases) according to random number table. Both groups were treated with ICS reduction based on the 2017 Gina Protocol. The treatment group was treated with Guben-Tiaoping Decoction for 12 weeks. The number of patients who were able to successfully complete the stepwise treatment after the end of the trial was calculated, the relevant indexes of Asthma Control Test (ACT), TCM Syndrome score, St. George’s respiratory questionnaire (SGRQ) and lung function were collected before and after treatment. Result:After 12 weeks’ treatment, the success rate of stepwise was 89.5% (34/38) in the treatment group and 78.9% (30/38) in the control group ( χ2=14.862, P=0.001). Meanwhile, the ACT score of the treatment group were better than those in the control group (Daytime symptoms t=-3.860, P=0.001, Dyspnea t=-2.007, P=0.40, Night waking t=-2.732, P=0.009, Reliever needed for symptoms t=-2.262, P=0.031, Control situation t=-6.994, P=0.001, Total Score t=-9.562, P=0.001). The score of TCM Syndrome in the treatment group was significantly lower than that of the control group ( P=0.001); PEF in treatment group [(6.92 ± 1.71) L vs. (5.84 ± 1.22) L; t=-2.880, P=0.005] was significantly better than that of the control group. Conclusion:Guben-Tiaoping Decoction is helpful to improve the success rate of ICS stepwise treatment and the life quality of patients.

To study the action, characteristics and expression of high methylation of p15(INK4B) and p16(INK4A) genes in multiple myeloma (MM), the sensitive methylation specific PCR method was employed to detect the hypermethylation of p15(INK4B) and p16(INK4A) in 24 patients with MM. Results showed that the methylation incidence of p15(INK4B) and p16(INK4A) genes were 70.8% (17/24) and 58.3% (14/24) in the MM patients, with the products of 148 bp and 150 bp fragments, respectively. The methylation of p15(INK4B) and p16(INK4A) genes were simultaneously happened in MM patients of plasmocytoma type with two cases at II phase and two cases at III phase. The simultaneous non-methylation of p15(INK4B) and p16(INK4A) genes were founded in five cases of MM patients, all of the tumor cells were of small plasmocyte type with mature differention. Conclusion suggested that there were high incidence of methylation of p15(INK4B) and p16(INK4A) genes in patients with MM. Hypermethylation can be detected in the early stage of disease, which was associated with its progress. It indicated a bad prognosis when methylation happended simultaneously in the two genes. Methylation of p15(INK4B) and p16(INK4A) genes may be related to the pathogeny of MM.

In order to explore the role of p15(INK4B) gene with highly methylated CpG island in the pathogenesis of leukemia, the expression levels of p15(INK4B) gene was detected in patients with AML and CML. Methylation-specific PCR (MSP) assay was employed in the experiments. The methylation incidence was 83.9% (26/31) in AML and 0% (0/28) in CML. The results showed that methylation of p15(INK4B) gene was the one of the major ways for inactivation of the gene, and the methylation could be appeared in clinical development of the disease and patients condition worsened. Methylation of p15(INK4B) did not occur and its function probably is normal in CML.

OBJECTIVETo detect mutations of ATP2A2 gene in a pedigree and a sporadic case with Darier disease (DD) and explore the underlying molecular mechanism.

METHODSClinical data of the pedigree and the sporadic case were collected. Genomic DNA was extracted from blood samples of four members from the pedigree (including three patients and one healthy member), the sporadic case and 100 healthy controls. PCR was performed to amplify all coding exons of the ATP2A2 gene. And the products were directly sequenced to detect mutations.

RESULTSA missense mutation c.1484C>T (p.S495L) in exon 12 was detected in all patients of the pedigree. For the sporadic case, a novel splicing mutation c.325-2A>G was detected at the junction between intron 4 and exon 5. The same mutations were not found in the 100 healthy controls.

CONCLUSIONMutations of the ATP2A2 gene may lead to the occurrence of DD in both familial and sporadic cases with DD.

Aged ; Alternative Splicing ; genetics ; Base Sequence ; Child ; DNA Mutational Analysis ; Darier Disease ; genetics ; Family Health ; Female ; Genetic Predisposition to Disease ; genetics ; Humans ; Male ; Mutation, Missense ; Pedigree ; Point Mutation ; Sarcoplasmic Reticulum Calcium-Transporting ATPases ; genetics