Research progress on effective components and mechanisms of anti-depressive Chinese medicine / 中国中药杂志

Since the pathogenesis of depression is complicated, the therapeutic effects of western medicine are poor accompanied by severe side effects. Chinese medicine has unique advantages in the treatment based on syndrome differentiation and contains many effective components against depression, including flavonoids, terpenes, phenylpropanoids, quinones, and alkaloids. These chemical components can delay the course of the disease, improve the curative effect, and reduce side effects of western medicine by regulating the biochemical abnormalities of monoamine neurotransmitters, brain tissue protein content, and internal environment as well as energy metabolism to make the treatment of Chinese medicine highlighted and recognized. This study systematically reviewed the effective components and mechanisms of anti-depressive Chinese medicine to inspire the rational development and utilization of new Chinese medicines against depression.

Role of Regional Network Management System in Disability Prevention for Newborn High-risk Infants / 中国康复理论与实践

Objective:To explore the feasibility of establishing a regional network management system to prevent and control the disability in high-risk infants. Methods:From July, 2015 to June, 2016, 1252 type B high-risk infants who born alive and registered in Lianyungang were divided into control group and experimental group by receiving network management system or not. The network high-risk infants management system was used to monitor the growth, diagnosis and early intervention of high-risk infants in the experimental group, while the control group was managed in the conventional way. A comprehensive physical examination and systematic assessment of 940 high-risk infants finally were conducted after two years. Their parents' compliance, developmental state, degree of dysplasia and function of dysplastic child were compared. Results:The compliance of parents was higher in the experimental group than in the control group (χ2 = 44.161,P < 0.001), as well as the outcome when the infants were two years old (χ2 = 204.340,P < 0.001). The younger they were found deviated and intervened, the better the outcome was (χ2 = 42.038,P < 0.001), and the less degree of dysplasia when they were two years old (χ2 = 10.508,P < 0.01). The deviation/abnormality condition was less in the experimental group than in the control group (χ2 = 17.446,P < 0.01). The development of functional area was better in the experimental group than in the control group (|t| > 2.206,P < 0.05), expect body structure (P > 0.05), in the infants with developmental deviation/abnormality. Conclusion:The establishment of network management system for high-risk infants can significantly improve the management compliance of parents and outcome of development of high-risk infants, to prevent disability.

Influence of dual antiplatelet therapy combined different dose of atorvastatin calcium on serum levels of MCP‐1 and VE‐cadherin in patients with acute cerebral infarction／ / 心血管康复医学杂志

To explore influence of dual antiplatelet therapy combined different dose of atorvastatin calci‐um on serum levels of monocyte chemoattractant protein (MCP)‐1 and vascular endothelial cadherin (VE‐cadherin) in patients with acute cerebral infarction atorvastatin routine dose group (ACI).Methods :A total of 119 ACI patients admitted in our hospital from 2016 to 2017 were randomly divided into atorvastatin routine dose group (n＝61) and large dose group (n＝58 ,40mg ,once／d) ,both groups received routine treatment and aspirin + clopidogrel for 30d. NIHSS ,serum levels of hsCRP ,TNF‐α ,IL‐8 ,MCP‐1 and VE‐cadherin ,carotid atherosclerotic plaque size and IMT before and after treatment ,and incidence of adverse reactions were measured and compared between two groups . Results :Compared with before treatment ,there was significant reduction in NIHSS score after 15d and 30d in two groups ,and those of 30d were significantly lower than those of 15d ,P＝0. 001 all ;compared with routine dose group after 15d and 30d ,there was significant reduction in NIHSS score [15d :(5.32 ± 1. 63 ) scores vs .(4. 13 ± 1.25 ) scores] in large dose group ,P＝ 0.001 ;Compared with routine dose group after 30d ,there were significant reduc‐tions in serum levels of hsCRP [(9.37 ± 1.85)mg／L vs.(6. 35 ± 1.94)mg／L] ,TNF‐α[(10. 15 ± 2.47)μg／L vs.(7. 44 ± 1.94)μg／L ] ,IL‐8 [(20. 35 ± 4.48 )μmol／L vs.(15. 14 ± 3. 61 ) μmol／L ] ,MCP‐1 [(234.54 ± 32. 53 ) ng／L vs. (185.46 ± 29. 47) ng／L] ,VE‐cadherin [(5. 32 ± 0. 49)mg／L vs.(4. 18 ± 0.54) mg／L] ,plaque size [(17.25 ± 3.14) mm2 vs.(14.13 ± 2.07)mm2 ] and IMT[(1.08 ± 0.25)mm vs.(0.85 ± 0. 17)mm] in large dose group ,P＝0.001 all. There was no significant difference in incidence rate of adverse reaction between two groups .Conclusion :Large dose atorvastatin combined dual antiplatelet therapy can significantly reduce serum levels of MCP‐1 and VE‐cadherin ,im‐prove brain function without increasing incidence rate of adverse reactions .

Significance of Peripheral Blood Lymphatic to Monocyte Ratio in the Progress of PGI-DLBCL / 中国实验血液学杂志

OBJECTIVE@#To explore the significance of lymphocyte to monocyte ratio (LMR) in the disease progress of primary gastrointestinal diffuse large B-cell lymphoma (PGI-DLBCL).@*METHODS@#The clinical data of 43 patients diagnosed as PGI-DLBCL in our hospital from January 2011 to December 2015 were collected, and the disease progress was followed up.@*RESULTS@#According to the ROC curve, the threshold value of LMR for 2 years PFS (%) of PGI-DLBCL patients was 2.6. Unvariate analysis showed that LMR (P＜0.05), large enclosed mass lesion (P＜0.01) and IPI (P＜0.05) were prognostic factors affecting PFS, the COX regression model multivariate analysis showed that LMR＜2.6 [ (risk ratio (RR)=3.083, 95%CI 1.828-8.313, P＜0.01], and large enclosed mass lesions (RR=2.718, 95%CI 1.339-6.424, P＜0.05) were the independent adverse prognostic factor for two years PFS.@*CONCLUSION@#Both LMR＜2.6 and large enclosed mass lesions relate with the progress of PGI-DLBCL.

Multivariate Analysis of Factors Influencing Recovery from Hemorrhagic Cystitis after Allo-HSCT / 中国实验血液学杂志

OBJECTIVE@#To analyze the incidence of hemorrhagic cystitis (HC) after allogeneic hematopoietic stem cell transplantation and the factors affecting HC, so as to provide clinical evidence for further treatment of HC.@*METHODS@#The HC of 113 patients after allogeneic hematopoietic stem cell transplantation in Affiliated Hospital of Xuzhou Medical University between the years 2014-2016 was analyzed respectively. All cases of HC were divided into HC group and non-HC(control) group. The follow-up time: from preeonditionig day to 180 d after transplantation. The 10 clinical parameters were selected for univariate analysis with COX regression analysis: sex, age (＜25 years and 25 years), primary disease, conditioning regimen with anti-thymoglobulin(ATG), sex-mismatch in recipients, haploidential HSCT, cytomegalovirus (CMV) viremia, EB viremia, graft-versus-host disease (GVHD), and primary disease relapse, the factors significant at the 0.1 level in univariate analysis should be further evaluated by multivariate analysis using a COX regression analysis. The difference was significant at P＜0.05 in multivariate analysis.@*RESULTS@#The HC occured in 31 of 113 patients (27.4%), with 5 cases of grade I (5.5%), 19 of grade II (16.8%), 5 of grade III (4.4%), and 2 of grade IV (1.8%). The median time of HC onset was 37 days (26-70 d) after transplantation. The median duration of HC was 14 days (5-55d). Univariate analysis showed that conditioning with anti-thymoglobulin (ATG) (RR=6.170, 95%CI: 1.875-20.306, P＜0.01), CMV viremia (RR=7.633, 95%CI：2.318-25.133) (P＜0.01), haploidentical HSCT (RR=0.307, 95%CI：0.137-0.686, P＜0.01), GVHD (RR=1.891, 95%CI：0.918-3.898, P＞0.05) were the risk factors for recovery from HC. The multivatiate analysis of above-mentioned risk factors with statistical significance showed that only CMV viremia (RR=4.770, 95%CI: 1.394-16.326, P＜0.05) was the indentified risk factor affecting the recovery from HC.@*CONCLUSION@#Monitoring CMV viremia and antivirotic treatment are effective measurs to prevent the occurrence of HC and promote the recovery from HC.

Overwintering surveillance of Culex pipiens pallens in Shandong Province / 中国血吸虫病防治杂志

Objective To understand the ecological habits of Culex pipiens pallens in Shandong Province in winter. Meth-ods From December 2015 to January 2016,the overwintering conditions of Cx.pipiens pallens were investigated in Shandong Province. Results In Shandong Province,in rural districts,the overwintering places of Cx. pipiens pallens were basements, wells and caves;and in urban areas,they were air raid shelters,holes of city walls,sewers and flower cellars.Conclusion In Shandong Province,the overwintering places of Cx.pipiens pallens are mainly basements and holes,which are under high tem-perature and humidity,and away from light.Its larvae cannot overwinter.

Rash and fever illness caused by herpes simplex virus type 1 needs to be distinguished from hand, foot and mouth disease / 病毒学报

An epidemic of rash and fever illnesses suspected of hand, foot and mouth disease (HFMD) occurred in Gansu Province of China in 2008, laboratory tests were performed in order to identify the pathogen that caused this epidemic. Eight clinical specimens collected from the 4 patients (each patient has throat swab and herpes fluid specimens) with rash and febrile illness, were inoculated onto RD and HEp-2 cells for virus isolation, and the viral nucleic acid was then extracted with the positive virus isolates, the dual-channel real-time reverse transcript-polymerase chain reaction (RT-PCR) was performed to detect the nucleic acid of human enterovirus (HEV) in the viral isolates at the same time. For the viral isolates with the negative results of HEV, a sequence independent single primer amplification technique (SISPA) was used for "unknown pathogen" identification. Totally, 6 viral isolates were identified as herpes simplex virus type 1 (HSV-1). Comprehensive analyses results of the clinical manifestations of the patients, epidemiological findings and laboratory test indicated that this epidemic of rash and febrile illness was caused by HSV-1. The differences among the gG region of 6 HSV-1 isolates at nucleotide level and amino acid level were all small, and the identities were up to 98. 8% and 97.9%, respectively, showing that this outbreak was caused by only one viral transmission chain of HSV-1. HSV-1 and other viruses that cause rash and febrile illnesses need differential diagnosis with HFMD. The etiology of rash and febrile illness is sometimes difficult to distinguish from the clinical symptoms and epidemiological data, the laboratory diagnosis is therefore critical.

Transmission and prevalence patterns of C4a evolutionary lineage of human enterovirus 71 circulating in mainland China, 2008-2010 / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To understand the evolutionary relationship between the C4a evolutionary lineage of human enterovirus 71 (HEV71) strains circulating in mainland of China during 2008-2010 and 2008 Fuyang strains and study the prevalence and transmission patterns of 2008 Fuyang strains.</p><p><b>METHODS</b>Download all the complete VP1 ( > or = 891 bp) or approximate complete VP1 (> or = 876 bp) gene nucleotide sequences from GenBank of HEV71 strains circulating in Mainland of China during 2008-2010. And analyze the phylogenetic relationship between Fuyang strains and other provinces' strains using the MEGA software, version 5.0.</p><p><b>RESULTS</b>All of the HEV71 isolates circulating in Mainland of China during 2008-2010 were clustered into evolutionary lineage C4a except for eight strains grouped in the genotype A and one isolate belongs to evolutionary lineage C4b; the homology analysis showed there were 96.5%-100% identity between C4a viruses circulating in mainland China during 2008-2010 and 2008 Fuyang strains, and they were evolved from C4b viruses of 1998. The transmission chains of Fuyang strains were mainly transmitted in Guangdong, Jiangsu, Shanghai, Hunan, Shandong provinces.</p><p><b>CONCLUSION</b>The predominant viruses circulating in Mainland of China during 2008-2010 were evolutionary lineage C4a of human Enterovirus 71; Fuyang transmission chains mainly distributed in southern of China and the Central China around Anhui provinces.</p>

Analysis of genetic characteristics of type II non-wild poliovirus in mainland China, 2010 / 病毒学报

To study the genetic characteristics of 123 type II non-wild polioviruses isolated from acute flaccid paralysis (AFP) cases in mainland China in 2010, provide the scientific basis for maintaining the "polio-free" status, and the switching use of polio vaccine for China. VP1 gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR) and the PCR products were then sequenced. The sequence results were analyzed with Sequencher 4.8, BioEdit 7.0.9 and MEGA 5.0. Of 65 strains, nt2909 was found to be a mutation hotspot, and also a neurovirulence determinant in VP1 region. During 2010, two vaccine-derived polioviruses (VDPVs) were isolated from Yunnan province, China and no wild poliovirus (WPV) was isolated. The epidemiological studies and laboratory results of the two VDPVs showed that they were newly discovered VDPVs because of the genetic difference from other VDPVs strains isolated in the world, implying the sensitive poliovirus surveillance network could timely detect the transmission of VDPVs and the importation of WPV.

The role of mir-221/222 in inhibiting endoplasmic reticulum stress-induced human hepatocarcinoma cell apoptosis / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the role of miR-221/222 in inhibiting endoplasmic reticulum stress-induced human hepatocarcinoma cells apoptosis.</p><p><b>METHOD</b>miR-221/222 mimics and inhibitors were used to mimic or block the function of endogenous miR-221/222 respectively. Western blot and flow cytometry were used to test the effects of miR-221/222 on cell cycle and apoptosis under endoplasmic reticulum stress in human hepatocellular carcinoma cells.</p><p><b>RESULTS</b>Endoplasmic reticulum stress resulted in miR-221/222 down-regulation in human hepatocellular carcinoma cells. miR-221/222 mimics and inhibitors inhibited and promoted respectively endoplasmic reticulum stress-mediated p27Kip1 induction. Moreover, p27Kip1 suppression not only resulted in reduction in the fraction of G1 phase cells, but also promoted the endoplasmic reticulum stress-mediated apoptosis in human hepatocellular carcinoma cells.</p><p><b>CONCLUSION</b>miR-221/222 were downregulated by endoplasmic reticulum stress in human hepatocellular carcinoma cells, which subsequently protected human hepatocellular carcinoma cells against endoplasmic reticulum stress-induced apoptosis through p27Kip1 regulation.</p>

Inhibition of evi1 expression by siRNA in HEL cell line / 中国实验血液学杂志

The aim of study was to investigate the inhibitory effect of small interfering RNA on evi1 gene expression and biological characteristics in HEL cells and its mechanism. 3 siRNA (siRNA-1, siRNA-2, siRNA-3) specific for evi1 gene were synthesized and transfected into HEL cells in vitro. Experiments were divided into test and control groups. MTT method was used to assay the inhibitory effect of siRNA on cell proliferation; semiquantitative RT-PCR was used to detect the expression of evi1 gene mRNA; the cell viability was determined by trypan blue dye test; the change of cell cycle and apoptosis of cells were analyzed by flow cytometry. The results showed that siRNA-1 had strongest effect, and inhibitory effect was most obvious at 48 hours after transfection. When the concentration of siRNA raised to 120 nmol/L, the inhibitory rate reached to the peak. The inhibitory rate of siRNA-1 on proliferation of HEL cells, relative expression level of evi1 mRNA and cell viability at 48 hours after transfection were 72.22 ± 2.80%, 27.31 ± 1.11% and 26.05 ± 2.49%, which had significant difference from other groups (p < 0.001). The siRNA resulted in arrest of cell cycle at G(0)/G(1) phase, the cell amount at S phase obviously decreased, the apoptotic rate of HEL cells obviously increased (p < 0.01). It is concluded that the siRNA specific for evi1 gene can suppress the proliferation of HEL cells, reduce the expression of evi1 mRNA, decrease the cell viability, arrest the cell cycle at G(0)/G(1) phase, suppress cell mitosis, and promote cell apoptosis.

Genetic characterization of coxsackievirus A16 isolated in Ningxia Hui Municipality in 2008 / 中华流行病学杂志

Objective To study the genetic characterization of coxsackievirus A16 (CVA16)strains isolated during an epidemic of hand, foot, and mouth disease (HFMD) in Ningxia Hui Municipality in 2008. Methods Clinical samples were collected from HFMD patients in Ningxia Hui Municipality and CVA16 strains were isolated by viral isolation methods. Reverse transcriptionpolymerase chain reactions (RT-PCR), specific for CVA16 were performed with these CVA16 strains.Entire VP1 coding region amplification and sequencing were then performed and finally phylogenetic tree was constructed among Ningxia CVA16 strains and CVA16 representative strains of known genotypes and subgenotypes. Results 70 Ningxia CVA16 strains were isolated from HFMD patients in Ningxia in 2008 and the homology of nucleotide and amino acid were 90.8%-100.0% and 98.9%-100.0%, respectively. Phylogenetic characteristics of the strains reconfirmed that they could be divided into two distinct genotypes-A and B. Genotype B could be further divided into the subgenotypes B1 and B2, while all the 70 Ningxia CVA16 strains belonged to the co-circulated clusters B1a and Blb within subgenotype B1, which belonged to 2 viral transmission chains.Conclusion Our results indicated that subgenotype B1 CVA16 strains continued to circulate over a wide geographic area of mainland China since the first reported episode in Shenzhen city in 1999. Like other CVA16 strains isolated elsewhere in China, both Bla and Blb evolution branches were co-circulating in Ningxia Hui Municipality. Based on the close phylgenetic and chronological relationship with CVA16 isolated in other countries and regions near China. Our data confimed that these strains co-evolved and co-circulated with those from neighboring countries and regions.

RT-PCR identification and genetic characterizations of VP1 region of human enterovirus 71 isolated in Beijing in 2008 / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study the genetic characterizations of VP1 region of Human enterovirus 71 (HEV71) isolated from clinical specimens of hand, foot and mouth disease (HFMD) patients in Beijing in 2008.</p><p><b>METHODS</b>285 clinical samples were collected from HFMD patients in hospitals and day-care centers in Chaoyang district. They were performed by reverse transcription-polymerase chain reaction (RT-PCR) specific for HEV71. 10 HEV71 isolates were selected for entire VP1 coding gene amplification and sequencing.</p><p><b>RESULTS</b>129 samples were RT-PCR positive, the positive rate is 45.26%. The homology of the nucleotide and the amino acid of the 10 strains were 94.6%-99.6% and 95.9%-100%. The phylogenetic tree revealed that 10 Beijing strains clustered within the C4a evolution branch of C4 subgenotype.</p><p><b>CONCLUSIONS</b>RT-PCR played an important role in identifying HFMD outbreak in Beijing in 2008. The HEV71 strains were all belong to C4a evolution branch of C4 subgenotype with several transmission chains, and it showed that C4 subgenotype HEV71 spread in mainland China widely after 1998. The molecular epidemiology surveillance and the research of genetic characterizations of HEV71 should be strengthened in mainland China.</p>

Cross-talk between PI3K/Akt and MEK/ERK pathways regulates human hepatocellular carcinoma cell cycle progression under endoplasmic reticulum stress / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the cross-talk between the PI3K/Akt and MEK/ERK pathways and its role in cell cycle regulation under endoplasmic reticulum stress in human hepatocellular carcinoma cells.</p><p><b>METHODS</b>PI3K inhibitor LY294002 and MEK inhibitor U0126 were used to block the PI3K/Akt and MEK/ERK pathways respectively, and constitutively activated Akt mutant construct was used to activate the PI3K/Akt pathway. Western blot was used to study the potential cross-talk between the PI3K/Akt and MEK/ERK pathways under endoplasmic reticulum stress in human hepatocellular carcinoma cells. the role of the cross-talk between the PI3K/Akt and MEK/ERK pathways in cell cycle regulation was investigated by using propidium iodide staining.</p><p><b>RESULTS</b>LY294002 not only blocked Akt activation efficiently but also increased ERK phosphorylation markedly under endoplasmic reticulum stress in SMMC-7721 and Hep3B cells. Furthermore, myr-Akt inhibited endoplasmic reticulum stress-mediated ERK phosphorylation. In contrast, MEK inhibitor U0126 had no effect on endoplasmic reticulum stress-induced Akt activation. It is notable that both myr-Akt overexpression and MEK inhibitor U0126 inhibited endoplasmic reticulum stress-induced G0/G1 phase arrest in SMMC-7721 cells.</p><p><b>CONCLUSION</b>Endoplasmic reticulum stress-induced Akt activation is mediated through PI3K and the PI3K/Akt pathway inactivation is involved in increased ERK activity in human hepatocellular carcinoma cells. The cross-talk between the PI3K/Akt and MEK/ERK cascades plays an important role in endoplasmic reticulum stress-induced human hepatocellular carcinoma cell cycle arrest.</p>

The epidemiologic and virological analysis of an outbreak of hand, foot, and mouth disease in Inner Mongolia in 2007 / 病毒学报

In 2007, an outbreak of hand, foot, and mouth disease (HFMD) occurred in Jungar Banner, Erdos city, Inner Mongolia Autonomous Region, China Fever, vesicular exanthema on the hands, feet, mouth, and buttocks were presented in most of the patients. Most of the patients were infants less than 5 years old, and an obvious peak period appeared in the disease outbreak. From 28 hospitalized patients, 23 stool specimens and 6 throat swab specimens were collected for enterovirus isolation, and 15 enteroviruses were isolated, 9 were identified as Human Enterovirus 71 (HEV71, the isolation rate is 31.03%) and 1 was identified as Coxsackievirus A16 (CVA16). According to the comprehensive analysis of clinical manifestation, epidemiology data and laboratory results, this outbreak was probably mainly caused by HEV71. The variability at nucleotide acid level and amino acid level among 9 HEV71 was relatively low, and the homology was more than 99.4% and 99.0% respectively, showing that this outbreak was caused by only one viral transmission chain. Phylogenetic analysis of 9 HEV71 strains isolated during this outbreak revealed that they all belonged to subgenotype C4, which has been continuously circulating in mainland China since its first reported occurrence in Shenzhen City in 1998. It was also suggested that subgenotype C4 HEV71 had a widely distribution and transmission in mainland China.

Outbreak of acute hemorrhagic conjunctivitis in Beijing City in 2007 caused by coxsackievirus A24 variant: molecular identification and phylogenetic analysis / 病毒学报

In 2007, an outbreak of acute hemorrhagic conjunctivitis (AHC) occurred in Beijing. In order to identify the etiology of this outbreak, 57 eye conjunctival swabs were collected from 57 outpatient patients, and detected for adenovirus, human enterovirus 70 (HEV70) and Coxsackievirus A24 variant (CVA24v) genes by using RT-PCR or PCR methods. The results showed that 38 were positive for CVA24v, the positive rate was 66.7%, but none was positive for HEV70 and adenovirus, showing that this outbreak was caused by CVA24v. 9 viral isolates were obtained from 57 clinical specimens by using viral isolation method, and all were identified as CVA24v by molecular typing method. All 9 CVA24v isolates were performed by VP1 sequencing, the results showed that except for strain 0744/BJ/CHN/2007, the variability at nucleotide acid level and amino acid level among other 8 CVA24v were relatively low, and the homologies were more than 99.6% and 100.0%, respectively; the homologies of nucleotide acid and amino acid between strain 0744/BJ/CHN/2007 and other 8 CVA24v were 96.8%-97.2% and 99.7%, respectively. Phylogenetic analysis of 9 CVA24v revealed that they represented the Clade 4 and Clade 5 in Group I, showed that this outbreak was caused by at least 2 viral transmission chains. Comparing to 3C region of CVA24v frequently used before, VP1 region was considered as the most rigorous target for molecular epidemiology study of CVA24v. To enhance the research of sero-epidemiology and molecular epidemiology of CVA24v and to know the genetic characterizations and molecular evolution of CVA24v are most important to prevent and control the outbreaks of AHC in China.

Genetic characterization of enterovirns 71 stains isolated in Ningxia province in 2008 / 中华流行病学杂志

Objective To study the genetic characterization of enterovirus 71 (EV71) strains isolated from specimens of patients with hand-foot-mouth disease (HFMD) in Ningxia province in 2008. Methods All the stool, throat swab and vesicle samples that collected from patients with HFMD were cultured. The positive isolates were identified by reverse transcriptase PCR (RT-PCR) with specific primers of EV71. Complete VP1 gene sequences (891 nucleotides) of 29 strains (part of 93 EV71 strains) were determined and compared with A, B and C genotype reference EV71 strains while EV71 China isolates by homogeneity and phylogenetic tree analyses. Results 215 strains of EV were isolated from 439 specimens. Results from RT-PCR indicated that 93 strains belong to EV71. Phylogenetic tree analysis revealed that the selected 29 stains were clustered with reference strains of C4 subgenotype. The nucleotide identity with C4 reference strains was 91.7%-99.4%. The amino acid homogeneity was 96.6%-100.0%. Conclusion The recently identified EV71 strains in Ningxia province belonged to subgenotype C4 which resembled to most of the isolates in China.

Two-dimensional electrophoresis analysis of hydrophobic membrane proteome in mouse hepatocarcinoma cell H22 / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To compare the membrane protein profile of mouse hepatocarcinoma cell H22 with that of normal liver cell.</p><p><b>METHODS</b>The membrane proteins in mouse hepatocarcinoma cell H22 and normal liver cell were extracted and their concentrations were determined by Bradford method. The proteins were separated by two-dimensional electrophoresis, and then stained with silver. The 2-DE maps were scanned and analyzed by Image Master 2D Platinum software. The differential expression protein spots were cut out from the gels, and the peptide fingerprinting was determined by MALDI-TOF-MS (Matrix-Assisted Laser Desorption/ Ionization Time of Flight Mass Spectrometry), followed by matching to Swiss-Prot protein database by Aldente software with experimental pI and MW data.</p><p><b>RESULTS</b>Compared to normal liver cells, 8 membrane proteins, including sulfatase-modifying factor 2, protein kinase C and casein kinase II substrate protein 3, sorting and assembly machinery component 50 homolog, macrophage scavenger receptor types I/II, uncharacterized protein C9 or f135 homolog, tight junction protein ZO-2, 3-hydroxy-3- methylglutaryl-coenzyme A reductase, and vacuolar protein sorting-associated protein 52 homolog were upregulated in H22 cells.</p><p><b>CONCLUSION</b>The membrane proteins involved in cell metabolism, proliferation, signal transduction, and skeleton, which are highly expressed in mouse hepatocarcinoma H22 cells, are probably related to the proliferation, invasion and migration of this tumor cell line.</p>

In vitro expression of hemophilia B gene mediated by lentivirus / 中华血液学杂志

<p><b>OBJECTIVE</b>To construct a three plasmids lentiviral vector containing canine coagulation factor IX (cFIX) gene with ubiquinone promoter (PUB) and observe the expression of cFIX gene.</p><p><b>METHODS</b>Lentivirus was generated by transient three-plasmid transfection, namely, the VSV-G envelope expression cassette, the delta NRF packaging plasmid and the PTK 164 plasmid. Viral particles were used to infect the target cell, third passage mesenchymal stem cells (MSCs) and 293T cell respectively at MOI 3: 1. The cFIX activity was detected in cultured cells with one-stage clotting assay.</p><p><b>RESULTS</b>The MSCs were obtained in vitro. The lentivirus infected MSCs and 293T cells all expressed the active factor IX with the activity of (26.30 +/- 2.10)% and (19.70 +/- 1.53)%, respectively, which are significantly higher than that of control (1.00 +/- 0.05)%.</p><p><b>CONCLUSIONS</b>The lentiviral vector of three plasmids with ubiquinone promoter (PUB) was constructed and can transfect the MSCs and 293T cells.</p>

Reevaluate the effect of G-480 point mutation that determines the neurovirulence of type I vaccine polioviruses / 病毒学报

Whole genome sequencing of 9 type I circulating vaccine-derived polioviruses (cVDPVs) isolated in Guizhou Province in China revealed that reverse mutations did not occur in G-480 and U-525 which are known as the most important neurovirulence determinate sites, while other known neurovirulence determinate sites such as A-2438, A-2795, C-6203 and G-7441 did revert to Mahoney type. 5 type I cVDPVs were selected for neurovirulence test on PVR-Tg21 transgenic mice which express human poliovirus receptor gene based on their different nucleotide sequences, they all showed higher neurovirulence than P1/Sabin strain, and the neurovirulence of CHN8184 and CHN8229-1. 1 were comparable to that of wild type P1/Mahoney. The neurovirulence of CHN8229-1.1, CHN8229-2 and CHN8229-3 presented a trend of decreasing, but still laid in high level. There were 7 nucleotide mutations between CHN8229-1.1 and CHN8229-2, and only 2 between CHN8229-2 and CHN8229-3 in their whole genomes, but the neurovirulence among them were relatively different, showing that there must be some unknown neurovirulence determinate sites among these mutations. Computer predicted RNA secondary structure of stem-loop V of the poliovirus 5' NCR of Guizhou type I cVDPVs was relatively stable. In the situation that no reverse mutation occurred in G-480, some type I cVDPVs already showed high neurovirulence nearly equal to P1/Mahoney, it meant that the effect of G-480 point mutation that determined neurovirulence of P1/Sabin strain has been overestimated, G-480 was not the only important site to determine neurovirulence in P1/Sabin strain, others also may play the very important role. More details are needed to elucidate the mechanism of attenuation in type I polioviruses.