1.Distribution in different Salmonella serovars and integration sites of Salmonella para-typhi C phage SPC-P1
Pan ZHAO ; Yihong PENG ; Qinghua ZOU
Journal of Peking University(Health Sciences) 2015;47(6):914-919
Objective:To determine the prevalence of Salmonella paratyphi C phage ( SPC-P1 ) in dif-ferent Salmonella serovars and to identify the integration sites in host genome. Methods: Based on the complete genome of SPC-P1 in S. paratyphi C RKS4594, 6 pairs of primers were designed and used to amplify the fragments of SPC-P1 in 11 S. typhi, 11 S. paratyphi A, 12 S. paratyphi B and 23 S. para-typhi C strains. At the same time, 100 complete genomes of Salmonella including 20 serovars available in National Center for Biotechnology Information ( NCBI) database were downloaded and aligned by Mauve 2. 3. 1 to determine the prevalence of SPC-P1 in these serovars. Primers were designed according to the integration sites of SPC-P1 in the genome of RKS4594 , and used to amplify ten strains having SPC-P1 in the genome. The PCR products were sequenced to investigate the integration sites of SPC-P1. Results:SPC-P1 was widely distributed in S. paratyphi C genome. In the study, 14 strains had all 6 fragments and 2 strains had 3-5 fragments. All the amplified fragments showed expected sizes. In contrast, in the ge-nomes of S. typhi, S. paratyphi A and S. paratyphi B, no or only 1-2 fragments could be amplified, and the sizes were smaller than expected. The results from Mauve showed that only in the genome of S. choleraesuis, which was a close relative of S. paratyphi C, there existed an almost complete genome of SPC-P1. The insertion site of SPC-P1 in all the ten S. paratyphi C strains tested was between pgtE and yfdC genes. Conclusion:SPC-P1 is a unique virulence factor of S. paratyphi C. It may play roles in the host range and pathogenicity of S. paratyphi C.
2.High-dose methylprednisolone effect on infantile spasms
Qinghua YANG ; Tianming JIA ; Liping ZOU ; Xiaoli ZHANG
Chinese Pediatric Emergency Medicine 2014;21(4):225-227
Objective To discuss high dose methylprednisolone therapy in infantile spasm.Methods Sixty-six children with infantile spasm of the Third Affiliated Hospital of Zhengzhou University from Nov 2002 to Nov 2012,adopted intravenous methylprednisolone with 20 mg/(kg· d) for 3 to 5 days continuously,then changed to oral prednisone with 1 ~ 2 mg/(kg.d),4 ~ 8 weeks after drug withdrawal.The total course of 8 ~ 12 weeks of clinical curative effect was retrospectively studied.Results (1) Of 66 cases,51 cases were effective,total effective rate was 77.2% ;28 cases achieved complete response,complete response rate was 42.4%.(2) The onset age had no influence on curative effect (P > 0.05).(3) The beginning ages of the patients to use methylprednisolone therapy had no effect on curative effect (P >0.05).(4) Course of curative effect:23 cases within 2 months,the effective rate was 91.3%,43 cases more than 2 months,the effective rate was 69.7%.Difference of effective rate between the two groups had statistical significance (x2 =3.958,P < 0.05).(5) Etiology of curative effect:50 cases of symptomatic,the effective rate was 78.0%,16 cases of cryptogenic,the effective rate was 75.0% ;compared two groups,the effective rates had no statistical significance(x2 =0.008 7,P > 0.05).(6) Long-term follow-up of 41 cases,14 cases had complete remission (34.2%),29 cases was effective(70.7%),3 cases had a recurrence (10.3%).Conclusion Application of methylprednisolone in the treatment for infantile spasm is simple,the recent control is effective,and it has a less adverse effect.The curative effect is related to the course,the shorter the duration,the better curative effect;and the long-term effect is good,the recurrence rate is low,the method can be recommended in clinical application.
3.Correlation between TCF7L2 gene polymorphism and genetic susceptibility in women with gestational diabetes mellitus
Xiling SHI ; Qinghua CAI ; Minyun ZOU ; Yousu SHEN
Chinese Journal of Obstetrics and Gynecology 2014;49(8):588-593
Objective To investigate the relationship between single nucleotide polymorphism (SNP) of transcription factor 7-like 2 (TCF7L2) at locus rs7903146,rs290487,rsl1196205,rs 12255372 and genetic susceptibility in women with gestational diabetes mellitus (GDM).Methods As a case-control study,100 pregnant women with GDM and 100 healthy pregnant women in the Maternal and Children Health Hospital of Jiangxi Province were recruited from January 2010 to July 2013.Clinical parameters,including body mass index (BMI),fasting insulin (FINS),fasting plasma glucose (FPG) and homeostatic model assessment-insulin resistance index (HOMA-IR) were measured after admission to hospital.Allelespecific PCR was used to analyze the SNP of TCF7L2 at locus rs7903146,rs290487,rs11196205,rs12255372.Results (1)The BMI,FPG,FINS and HOMA-IR in GDM group were(27.4±3.0) kg/m2,(5.6±1.0) mmol/L,(6.2±3.4) mU/L and 1.8± 1.0,and were (24.2±2.9) kg/m2,(5.3±0.8) mmol/L,(4.5±2.8) mU/L,1.2± 0.8 in the control group,respectively.The differences had statistically significance (P<0.05).(2)The SNP of TCF7L2 gene,locus rs7903146 were CC,CT and TT genotype; the SNP of locus rs290487 were CC,CT and TT genotype; and the SNP of locus rs1 1196205 were GG and CC genotype; while the SNP of locus rs12255372 was GG genotype.(3)The distribution frequencies of genotype CC,CT and TT at locus rs7903146 in the GDM group were 40% (40/100),36% (36/100) and 24% (24/100),respectively.While in the control group,they were 55% (55/100),38% (38/100) and 7% (7/100),respectively.The frequencies of C and T allele of rs7903146 were 58%and 42% in the GDM group,and in the control group they were 74% (148/200) and 26% (52/200).The differences of genotype distribution and C/T allele frequency of rs7903146 between the two groups were statistically significant (P<0.05).(4)The distribution frequencies of genotype CC,CT and TT at locus rs290487 in the GDM group were 12 % (12/100),36 % (36/100) and 52% (52/100),and were 16% (16/100),34% (34/100) and 50% (50/100) in the control group.The frequencies of C and T allele of rs290487 were 30% (60/200) and 70% (140/200) in the GDM group,and were 33% (66/200) and 67% (134/200) in the control group.There was no difference of genotype distribution and C/T allele frequency of rs290487 between the two groups (P>0.05).(5)The distribution frequencies of genotype GG and CC at locus rs1 1196205 in the GDM group were 99% (99/100) and 1% (1/ 100),while those in the control group were 100%(100/100) and 0%.The frequency of G and C allele of rs1 1196205 were 99%(198/200) and 1%(2/200) in the GDM group,while in the control group were 100% (200/200) and 0.There was no difference of genotype distribution and G/C allele frequency of rs11196205 between the two groups (P>0.05).(6)The distribution frequencies of genotype GG at locus rs12255372 were 100%(100/100) in both the GDM group and the control group.The frequencies of G allele of rs12255372 were 100% (200/200) in both the GDM group and the control group.There was no difference of genotype distribution and G allele frequency of rs12255372 between the two groups (P>0.05).(7)After adjusting for age,gestational age,BMI,FPG and FINS,pregnant women with TT genotype at locus rs7903146 were more likely to have hyperglycemia compared with the C allele carriers (OR=2.77,95% CI:1.03-7.57,P<0.05).Conclusions The polymorphism of locus rs7903146 in TCF7L2 gene may be associated with genetic susceptibility in women with GDM.TT genotype is likely to be risk factor in the pathogenesis of GDM.
4.Role of oxygen free radicals in hypertension and cardiovascular remodeling of nitric oxide-deficient rats
Qi CHEN ; Yanqing WU ; Xiaoshu CHENG ; Bin ZOU ; Qinghua WU ; Hai SU ; Juxiang LI
Chinese Journal of Tissue Engineering Research 2006;10(44):198-201
BACKGROUND: Our previous study showed that the activation of local renin-angiotensin system in heart and vessels contributed to hypertension and cardiovascular remodeling. However, whether oxygen free radical plays an important role in this process is still unclear. OBJECTIVE: To investigate the interventional effects of Ebselen, a kind of anti-oxidative drug, on rats administered by Nw-Nitro-L-arginine methyl easter (L-NAME) (L-NAME), inhibitor of nitric oxide synthase (NOS) for a long term, and probe into the role of oxygen free radicals (OFR) in hyper- tension and cardiovascular remodeling of NO-deficient rats. DESIGN: A randomized grouping and controlled animal trial. SETTING: Department of Cardiology, the Second Affiliated Hospital of Nanchang University, Institute of Cardiology, Nanchang University. MATERIALS: The experiment was completed from January 2002 to March 2003 at the Animal Experimental Lab, Institute of Cardiology, Nanchang University and the Key Molecular Medical Lab of Jiangxi Province. Twenty-four Wistar rats were divided to three groups according to the random number table method: normal control group (n=8), L-NAME group (n=8), L-NAME + Ebselen group (n=8). METHODS: ①Normal control group: The rats could eat and drink routinely, and they were administrated by skim milk ball (net weigh = 4 g) before feed every night. ② L-NAME group: The rats received L-NAME in the dose of 50 mg/kg mixed in one skim milk ball everyday before feed every night. ③ L-NAME + Ebselen group: The rats were admin istered by one skim milk ball (net weigh = 2 g) mixed with L-NAME (50 mg/kg) and one skim milk ball (net weigh = 2 g) mixed with ebselen (30 mg/kg). The systolic blood pressure (SBP) was detected before LNAME was given and at the ends of the 1st, 2nd, 4th,6th and 8th weeks respectively. The rats were killed under anesthesia at the end of the 8th week, the plasma and homogenate of myocardium of apex were taken to detect the biochemical indexes, the other heart tissues were used for the histological detections. MAIN OUTCOME MEASURE: ① Dynamic changes of blood pressure were observed. The levels of NO and malondialdehyde (MDA), activities of angiotensin-converting enzyme (ACE) and superoxide dismutase (SOD) in plasma and myocardium of apex were measured. The production of superoxide anion and the levels of angiotensin Ⅱ type 1 receptor (AT1R) protein expression in myocardium of apex were determined. ③ The pathomor- phological indexes were determined. RESULTS: All the 24 rats were involved in the analysis of results without deletion. ① SBP: At the ends of the 1st, 2nd, 4th 6th and 8th weeks, SBP elevated gradually in the L-NAME group, which were significantly higher than those in the control group at the same time (P < 0.05-0.01). At the end of the 8th week, the SBP was significantly lower in the L- NAME + Ebselen group than in the L-NAME group (P < 0.05). ② Bio chemical indexes in plasma and homogenate of myocardium of apex: The NO level and SOD activity in myocardial tissue were significantly lower in the L-NAME group than in the normal control group (P < 0.05-0.01), but significantly higher in the L-NAME + Ebselen group than in the L- NAME group (P < 0.05-0.01). The production of superoxide anion, ACE activity and level of AT1R expression were all significantly higher in the L-NAME group than in the normal control group (P < 0.05), but signifi- cantly lower in the L-NAME + Ebselen group than in the L-NAME group. ③ Pathomorphological indexes: The ratio of cardiac mass to body mass, thickness of left ventricle and ratio of thickness to lumen diameter of small artery were all significantly higher in the L-NAME group and L- NAME + Ebselen group than in the normal control group (P < 0.05-0.01), and the thickness of left ventricle and ratio of thickness to lumen diameter of small artery were significantly lower in the L-NAME + Ebselen group than in the L-NAME group (P < 0.05) CONCLUSION: Ebselen, an anti-oxidative drug, enable to attenuate the development of hypertension and cardiovascular remodeling in NO-deficient rats. By activating renin-angiotensin system (RAS), OFR may accelerate the formation of hypertension and cardiovascular remodeling, and the increased production of OFR may contribute to the development of cardiovascular remodeling. It is indicated that RAS may play an important role in the development of hypertension and cardiovascular remodeling induced by NO-deficiency
5.Construction and identification of p4CCL20-ZsGreen1-DR eukaryotic expression vector
Yong WANG ; Zhizhong WANG ; Bing ZHONG ; Heng WANG ; Qinghua ZOU ; Yu CHEN
Chinese Journal of Tissue Engineering Research 2011;15(41):7719-7722
BACKGROUND: It is necessary to establish a high throughput screening system for anti -inflammatory drugs for rheumatoid arthritis.OBJECTIVE: To construct an eukaryotic expression vector p4CCL20-ZsGreen1-DR with the NF-kB cis-acting element 4×CCL20motif as an enhancer, SV40 as a promoter, and ZsGreen1-DR as a reporter gene.METHODS: The target fragment SV40 was PCR amplified using PGL2-control plasmid as a template. KpnⅠ/Bam HⅠ restriction sites were introduced into the flank of the target fragment. Then, pSV40-ZsGreen1-DR vector was constructed by cloning the target fragment into pZsGreen1-DR plasmid. Finally, p4CCL20-ZsGreen1-DR plasmid was constructed by cloning the double strand DNA of 4×CCL20 motif (with BglⅡ and EcoRⅠ sticky ends at the 5’ and 3’ terminus, respectively) into the corresponding restriction sites of pSV40-ZsGreen1-DR vector (upstream of SV40 promoter).RESULTS AND CONCLUSION: DNA sequencing demonstrated successful construction of p4CCL20-ZsGreen1-DR plasmid.The construction of p4CCL20-ZsGreeR plasmid might be useful to establish a high throughput screening system for anti -inflammatory drugs.
6.Risk factors and treatment efficiency for lung cancer patients with venous thromboembolism
Quanfang CHEN ; Wei WANG ; Xiaoying ZOU ; Zhian LING ; Yanbin WU ; Jinliang KONG ; Banghao XU ; Qinghua DU
The Journal of Practical Medicine 2014;(6):891-894
Objective To investigate the risk factors and treatment efficiency for lung cancer patients with venous thromboembolism (VTE). Methods Total 282 cases of lung cancer patients with VTE were enrolled into two groups , including the VTE group and the non-VTE group , for comparation analysis based on a series of clinical data. Results The occupation rate of adenocarcinoma and Ⅳ period were 65.28% and 87.50% in VTE group, respectively, higher than those of 51.43% and 75.71% in the non-VTE group. The increased rate of blood viscosity and d-dimer respectively were 65.28% and 70.83%, higher than those of 51.43% and 56.67% in the non-VTE group, with significant differences (P < 0.05, respectively). Result of logistic regression analysis showed that tumor stage , d-dimer levels , smoking , age , and blood viscosity levels were highly correlated with venous thrombosis in patients with lung cancer, and the OR value among them was 3.802, 2.339, 5.814, 3.875 and 6.404, respectively, with significant differencees (P < 0.05, respectively). Conclusions Lung adenocarcinoma with stage Ⅳ, smoking , age and increase of blood viscosity and d-dimer were the important risk factors for VTE in patients with lung cancer chemotherapy. Timely assessment of risk factors and early anticoagulation therapy in lung cancer patients with venous thromboembolism associated with VTE can improve the treatment efficacy and reduce the complications.
7.The relationship between social anxiety and self-esteem, self-awareness in grade 4-6 primary students
Zhili ZOU ; Pinhong CHEN ; Huaqing MENG ; Hui WANG ; Qinghua LUO ; Yixiao FU ; Lian DU ; Haitang QIU ; Tian QIU ; Hua HU
Chinese Journal of Behavioral Medicine and Brain Science 2012;21(5):436-439
ObjectiveTo explore the status of social anxiety and study the relationship between social anxiety and self-consciousness,self-esteem among primary students.MethodsUsing stratified randomization sampling method,568 primary students in grade 4 ~ 6 were investigated with social anxiety scale for children( SASC ),self-consciousness scale for children(PHCSS) and self-esteem scale(SES).Results①The score of SASC in the group of primary students was obviously higher than that of Chinese norm (P<0.01).②The SASC and negative evaluation scores in the female were higher than the male (P < 0.05 ),the negative evaluation scores in the sixth grade (3.22 ± 2.82) was higher than the fifth grade (2.53 ± 2.76,P < 0.01 ) and fourth grade ( 2.54 ± 2.68,P< 0.05 ).The primary students in migrant workers elementary school,the single parent family and the relatively low income family had higher social anxiety scores than others(P<0.05 ).③The SASC and two major factors were significantly negative correlated with the self-consciousness and each subscale( r =-0.202 ~ -0.511,P < 0.01 ),and positive correlated with the self-esteem( r =0.282,0.270,0.305,P < 0.01 ).④)The anxiety and gregariousness had significant prediction function of 25.5% on negative evaluation,and 28.2% on SASC,anxiety,gregariousness,intelligence and school situation had significant prediction function of 20.4% on social avoidance and distress.ConclusionsPrimary students in grade 4 ~ 6 existed social anxiety.There are significant differences for their level of anxiety in different gender,grade,elementary school,family structure,monthly income.And social anxiety is closely related to self-consciousness and self-esteem.
8.Theory and practice regarding risk assessment of public health emergencies in Armed Forces
Hailong SUN ; Wen ZOU ; Wenyi ZHANG ; Shenlong LI ; Yong WANG ; Yanan LI ; Qinghua LI ; Yue LI ; Yansong SUN
Military Medical Sciences 2014;(1):10-13
Risk assessment is critical to responses to public health emergencies .Practices and researches concerning risk assessment of public health emergencies in China′s military are still in the early stage .This paper summarizes the rele-vant concepts , categorization , assessment methods , and practices .Based on management requirements of risk assessment , suggestions are raised about how to strengthen the risk assessment capability of Armed Forces by formulating management rules, perfecting assessment methods and developing assessment tools .
9.Effect of livin-modified BM-MSCs transplantation on cardiac function fol-lowing acute myocardial infarction in a rat model
Bing ZOU ; Junping XIE ; Qinghua WU ; Shoulin CHEN ; Lumin XIAO ; Hai SU ; Kui HONG ; Yanqing WU ; Xiaoshu CHENG
Chinese Journal of Pathophysiology 2016;32(3):539-543
[ ABSTRACT] AIM:To study the effect of livin gene-modified bone marrow mesenchymal stem cells ( BM-MSCs) transplantation on the cardiac function following acute myocardial infarction in a rat model and the expression of livin , caspase-3, caspase-7 and caspase-9 in the livin gene-modified BM-MSCs.METHODS: The MSCs were obtained by the whole bone marrow culture method , and the apoptosis of the MSCs after infection with adenovirus vector carrying enhanced green fluorescent protein ( EGFP) gene and livin recombinant vector ( rAd-livin) were detected by flow cytometry .The ex-pression of livin, caspase-3, caspase-7 and caspase-9 was detected by Western blot .After permanent left anterior descend-ing artery occlusion , the rats were randomized to receive intramyocardial injection of DMEM without cells ( vehicle group ) , or containing MSCs ( MSCs group ) , MSCs ( EGFP ) ( rAd-control/MSCs group ) or MSCs ( livin ) ( rAd-livin/MSCs group).Left ventricular systolic pressure (LVSP), left ventricular end-diastolic pressure (LVEDP), the maximum in-creased rate of left ventricular pressure ( -dp/dtmax ) and the maximum decline rate of left ventricular pressure ( +dp/dtmax ) were recorded for evaluating the cardiac functions .RESULTS: The apoptosis of rAd-livin/MSCs was significantly decreased as compared with MSCs and rAd-control/MSCs (P<0.05).Meanwhile, the expression of caspase-3, caspase-7 and caspase-9 was significantly downregulated as compared with the other 2 groups ( P<0.05 ) .The cardiac function in rAd-livin/MSCs group was significantly improved as compared with DMEM group , and those in the other 2 groups got the similar results, but the function in rAd-livin/MSCs group was better improved .Meanwhile, the number of surviving cells in rAd-livin/MSCs group was significantly improved as compared with the other 2 groups .CONCLUSION:The apoptosis of MSCs is decreased after rAd-livin transfection, and the expression of caspase-3, caspase-7 and caspase-9 is also significant-ly downregulated while the expression of livin is significantly upregulated .Transplantation of livin-modified BM-MSCs by lentiviral vector results in better prognosis for treating myocardial infarction by enhancing cell survival .
10.A familial case of Blau syndrome caused by dual NOD2 genetic mutations
Hong LI ; Zhijun JIANG ; Qinghua HU ; Xiaohui LIU ; Zheng ZOU
Chinese Journal of Applied Clinical Pediatrics 2018;33(20):1560-1563
Objective To provide the diagnostic proof for a suspected Chinese family with BS,and NOD2 gene mutation types and clinical features were analyzed in this study.Methods Nine members (4 males and 5 females) of this family were enrolled.To clarify the genotype,the whole exome sequencing by next-generation sequencing from the proband and his parents was performed,and all members were subjected to Sanger sequencing.For the newly discovered NOD2 missense mutation,its pathological predictions were conducted online by adopting polyphen software.Clinical data of affected cases diagnosed by NOD2 analysis were collected to analyze illustrate the clinical features.Results (1)The proband of the family was a 5-year-old Chinese Han boy,who had the clinical triad of dermatitis,polyarthritis and uveitis.The body temperature and C-reactive protein (CRP) was normal.Besides the proband,2 members were diagnosed as BS by means of NOD2 analysis.The coexistence of 2 missense mutations was detected.One novel mutation was c.1981G > C,p.A661 P,and another previously reported one was c.2006A > G,p.H669A.(2) Mutations identified in the male proband were inherited from his father.Tracing the other pedigree members,it was disclosed that his grandmother had the heterozygous dual NOD2 mutations.The proband displayed a phenotype featuring the symptom triad of granulomatous dermatitis,symmetric arthritis,and recurrent uveitis,with normal temperature and CRP level.Conclusions The coexistence of A661P and H669A mutations in NOD2 caused BS in a Chinese pedigree,which derived from the proband's grandmother.This is the first report of A661P mutation in NOD2 in a Chinese pedigree of this disease.The proband has multi hydatoncus surrounding multi-joints,but no persistent fever and no elevated CRP,which may help to differentiate BS from other inherited autoinllammatory diseases in clinical settings.