Advances in basic research on choline and central nervous system development and related disorders / 中华预防医学杂志

Choline is an essential nutrient that plays an integral role in all stages of the life cycle, with increasing interest in the relationship between choline and neurodevelopment. Choline is a major component in the synthesis of phospholipids, phosphatidylcholine and sphingolipids, and is an essential nutrient for methyl metabolism, acetylcholine synthesis and cell signaling. Choline plays an important role in neurogenesis and neural migration during fetal development, potentially influencing the development and prognosis of neurological disorders, but its mechanism of action is not yet clear. This article reviews the source and metabolism of choline, the effects and mechanism of choline on neurodevelopment and central nervous system related disorders.

Effect of different maintenance doses of caffeine citrate on ventilator weaning in very preterm infants with respiratory distress syndrome: a prospective randomized controlled trial / 中国当代儿科杂志

OBJECTIVES@#To study the effect of different maintenance doses of caffeine citrate on the success rate of ventilator weaning in very preterm infants (gestational age of ≤32 weeks) with respiratory distress syndrome (RDS).@*METHODS@#A total of 162 preterm infants with RDS who were admitted to the hospital from January 2016 to December 2018 were enrolled in this prospective trial. These infants had a gestational age of ≤32 weeks and required invasive mechanical ventilation. They were randomly divided into a high-dose caffeine group and a low-dose caffeine group, with 81 infants in each group. Within 6 hours after birth, both groups were given caffeine at a dose of 20 mg/kg. After 24 hours, the high- and low-dose caffeine groups were given caffeine at a maintenance dose of 10 mg/kg and 5 mg/kg, respectively. The two groups were compared in terms of re-intubation rate within 48 hours after ventilator weaning, durations of ventilation and oxygen therapy, enteral feeding, weight gain, and the incidence rates of complications and adverse reactions during hospitalization.@*RESULTS@#The high-dose caffeine group had a significantly lower re-intubation rate within 48 hours after ventilator weaning than the low-dose caffeine group (@*CONCLUSIONS@#A high maintenance dose of caffeine can safely and effectively reduce the incidence rate of apnea after ventilator weaning and the failure rate of ventilator weaning in RDS preterm infants with a gestational age of ≤32 weeks, and therefore, it holds promise for clinical application.

Neuropsychological development of late preterm infants and early term infants at the age of 1 year: a follow-up study / 中国当代儿科杂志

OBJECTIVE@#To study the level of neuropsychological development in late preterm infants and early term infants at the age of 1 year.@*METHODS@#A total of 1 257 children with a corrected age of 1 year were enrolled as subjects. According to gestational age at birth, they were divided into an early preterm group (28-33 weeks), a late preterm group (34-36 weeks), an early term group (37-38 weeks), and a full-term group (39-41 weeks). Gesell Developmental Schedules were used to assess the neuropsychological development of the children, and the groups were compared in terms of neuropsychological development at the age of 1 year.@*RESULTS@#There were significant differences in the developmental quotients of the five functional areas (adaptability, gross motor, fine motor, language and social ability) between the four groups at the age of 1 year (P<0.05), and the full-term infants had the highest development quotients, followed by the early term infants, the late preterm infants, and the early preterm infants (P<0.05). The full-term infants had the lowest rate of developmental delay in each functional area, while the early preterm infants had the highest rate (P<0.05). Compared with the full-term infants, the early term infants had a higher risk of developmental delay in adaptability (OR=1.796, P<0.05), and the late preterm infants had a higher risk of developmental delay in adaptability (OR=2.651, P<0.05) and fine motor (OR=2.679, P<0.05), while the early preterm infants had a higher risk of developmental delay in adaptability (OR=4.069, P<0.05), fine motor (OR=3.710, P<0.05), and social ability (OR=3.515, P<0.05).@*CONCLUSIONS@#The risk of neuropsychological developmental delay decreases with the increase in gestational age in children at the age of 1 year, with a dose-response effect. There are varying degrees of developmental delay in early term infants and late preterm infants, and health care follow-up for early term infants and late preterm infants should be taken seriously.

Effect of CXCL12/CXCR4/MMP-2 signaling pathway on biological behavior in triple negative breast cancer / 临床与实验病理学杂志

Purpose To investigate the effect of CXCL12/CXCR4 and MMP-2 on the biological behavior of triple-negative breast cancer.Method CXCR4-siRNA and pcDNA3.1-CXCR4were transient transfected of in MDA-MB-231 cell line by liposome,RNA and protein were extracted.The mRNA and protein expressions of CXCR4,CXCL12 and MMP-2 were detected by RT-PCR and Western blot respectively.And the migration and proliferation of the transfected cells were detected by Wound-healing assay and MTF assay.Result The experiment of RT-PCR and Westen blot demonstrated that CXCR4 mRNA and protein level expression declines after the expression CXCR4 was down-regulated (P ＜ 0.01),CXCL12 mRNA and protein level expression increased and MMP-2 mRNA and protein level expression increased after the expression CXCR4 was down-regulated (P＜0.01),MTT assay and Wound-healing assay results showed that the proliferation ability of 72 h cells was decreased,and the wound healing was slow.The experiment of RT-PCR and Westen blot demonstrated that CXCR4 mRNA and protein level expression increased after the expression CXCR4 was up-regulated (P ＜0.01).The CXCL12 mRNA and protein level expression declined and MMP-2 mRNA and protein level expression increased after the expression CXCR4 was up-regulated (P ＜0.01),MTT assay and wound-healing assay results showed that the proliferation ability of 72 h cells was increased,and the wound healing was accelerated.Conclusion CXCL12/CXCR4/ MMP-2 signaling pathway is activated in the triple-negative breast cancer MDA-MB-231 cells,and the inhibition of CXCR4 can reverse the proliferation and migration of malignant biological process.

Depression, anxiety, and quality of life in paroxysmal kinesigenic dyskinesia patients / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Paroxysmal kinesigenic dyskinesia (PKD) is a rare movement disorder characterized by recurrent dystonic or choreoathetoid attacks triggered by sudden voluntary movements. Under the condition of psychological burden, some patients' attacks may get worsened with longer duration and higher frequency. This study aimed to assess nonmotor symptoms and quality of life of patients with PKD in a large population.</p><p><b>METHODS</b>We performed a cross-sectional survey in 165 primary PKD patients from August 2008 to October 2016 in Rui Jin Hospital, using Symptom Check List-90-Revised (SCL-90-R), World Health Organization Quality of Life-100 (WHOQoL-100), Self-Rating Depression Scale, and Self-Rating Anxiety Scale. We evaluated the differences of SCL-90-R and WHOQOL-100 scores in patients and Chinese normative data (taken from literature) by using the unpaired Student's t-test. We applied multivariate linear regression to analyze the relationships between motor manifestations, mental health, and quality of life among PKD patients.</p><p><b>RESULTS</b>Compared with Chinese normative data taken from literature, patients with PKD exhibited significantly higher (worse) scores across all SCL-90-R subscales (somatization, obsessive-compulsive, interpersonal sensitivity, depression, anxiety, hostility, phobic anxiety, paranoid ideation, and psychoticism; P= 0.000 for all) and significantly lower (worse) scores of five domains in WHOQoL-100 (physical domain, psychological domain, independence domain, social relationship domain, and general quality of life; P= 0.000 for all). Nonremission of dyskinesia episodes (P = 0.011) and higher depression score (P = 0.000) were significantly associated with lower levels of quality of life. The rates of depression and anxiety in patients with PKD were 41.2% (68/165) and 26.7% (44/165), respectively.</p><p><b>CONCLUSIONS</b>Depression, anxiety, and low levels of quality of life were prevalent in patients with PKD. Co-occurrence of depression and anxiety was common among these patients. Regular mental health interventions could set depression and anxiety as intervention targets. Considering that the motor episodes could be elicited by voluntary movements and sometimes also by emotional stress, and that symptoms may get worsened with longer duration and higher frequency when patients are stressed out, intervention or treatment of depression and anxiety might improve the motor symptoms and overall quality of life in PKD patients.</p>

Incidence of late-onset sepsis in very low birth weight and extremely low birth weight infants and risk factors for late-onset sepsis / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the incidence of late-onset sepsis (LOS) in very low birth weight (VLBW) and extremely low birth weight (ELBW) infants in the neonatal intensive care unit (NICU) and the risk factors for LOS.</p><p><b>METHODS</b>A retrospective analysis was performed for the clinical data of all VLBW and ELBW infants who were hospitalized in the NICU between January 2011 and December 2013. According to the presence or absence of LOS, these infants were divided into LOS group and non-LOS group. The incidence and mortality rates of LOS, common pathogenic bacteria, and risk factors for LOS were analyzed.</p><p><b>RESULTS</b>Of the 226 VLBW and ELBW infants, 117 (51.8%) developed LOS, among whom 45 had a confirmed diagnosis of LOS and 72 had a clinical diagnosis of LOS. The LOS group had a significantly higher mortality rate than the non-LOS group [13.7% (16/117) vs 4.6% (5/109); P<0.05]. Bacterial culture found 51 strains of pathogenic bacteria, among which 32 (63%) were Gram-negative bacteria, 16 (31%) were Gram-positive bacteria, and 3 (6%) were fungi. The multivariate logistic regression analysis showed that gestational age, small for gestational age (SGA), duration of parenteral nutrition, peripherally inserted central catheter (PICC) placement, and mechanical ventilation were independent risk factors for LOS in VLBW and ELBW infants (OR=0.84, 1.59, 1.34, 3.11, and 4.55 respectively; P<0.05).</p><p><b>CONCLUSIONS</b>LOS has high incidence and mortality rates in VLBW and ELBW infants. Common pathogenic bacteria of LOS are Gram-negative bacteria. Low gestational age, long duration of parenteral nutrition, SGA, PICC placement, and mechanical ventilation may increase the risk of LOS in VLBW and ELBW infants.</p>

Early intellectual developmental outcome of late preterm infants / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the early intellectual developmental outcome of late preterm infants.</p><p><b>METHODS</b>A total of 106 late preterm infants with a gestational age of 34-36weeks who were admitted to the neonatal ward between January 2012 and January 2015, cured, discharged, and regularly followed up at the outpatient service for high-risk children were enrolled as the preterm group. A total of 120 healthy full-term infants during the same period were randomly selected as the term group. Neonatal behavioral neurological assessment (NBNA) was performed for late preterm infants at a corrected gestational age of 40 weeks and full-term infants at a gestational age of 40 weeks. The Gesell Developmental Scale was used for late preterm infants at a corrected age of 3, 6, and 12 months and full-term infants at an age of 3, 6, and 12 months.</p><p><b>RESULTS</b>The preterm group had an NBNA score of <37 and a significantly lower NBNA score than the term group (P<0.05). At the corrected age of 3 months, the preterm group had significantly lower scores of gross motor, fine motor, and social competence than the term group (P<0.05). At the corrected age of 6 months, the preterm group had significantly lower scores of adaptability, gross motor, and fine motor than the term group (P<0.05). At the corrected age of 12 months, the preterm group had significantly lower scores of adaptability, gross motor, and social competence than the term group (P<0.05).</p><p><b>CONCLUSIONS</b>Late preterm infants have early intellectual developmental delay. It is necessary to perform neurodevelopmental monitoring for late preterm infants.</p>

Risk factors for extrauterine growth retardation at discharge in premature infants / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the incidence and risk factors for extrauterine growth retardation (EUGR) at discharge in premature infants.</p><p><b>METHODS</b>A retrospective analysis was performed on 596 premature infants who were admitted to the neonatal intensive care unit between 2006 and 2010. These subjects were classified into EUGR (n=217) and non-EUGR groups (n=379) based on the body weight at discharge. The risk factors for the occurrence of EUGR were studied by multivariate logistic regression analysis.</p><p><b>RESULTS</b>Based on the body weight, length, and head circumference, the incidence of EUGR at discharge was 36.4% (217 cases), 42.0% (250 cases), and 22.8% (136 cases), respectively. Low gestational age, low birth weight, intrauterine growth retardation (IUGR), delayed enteral feeding and complications of the respiratory system were identified as risk factors for EUGR (OR=6.508, 14.522, 5.101, 1.366, and 1.501, respectively).</p><p><b>CONCLUSIONS</b>The incidence of EUGR might be greatly decreased by strengthening the perinatal care, reducing the incidence of premature delivery and IUGR, undertaking early enteral feeding, and actively preventing postnatal complications.</p>

Prevalence and related risk factors of anxiety and depression among Chinese college freshmen / 华中科技大学学报（医学）（英德文版）

Anxiety, depression, and even suicidal ideation are becoming the most common mental health problems affecting Chinese college students. The present study investigated the prevalence of mental health problems and their predictors in a sample of 1048 Chinese college freshmen from Shanghai. We used following brief screening instruments to measure symptoms of anxiety and depression, as well as self-control and suicidal ideation: the Patient Health Questionnaire (PHQ-9), the Generalized Anxiety Disorder scale (GAD-7), a mental health and mental health knowledge questionnaire (MK), a mental disease-related attitude questionnaire (MA), questionnaires about the knowledge of psychological services and utilities, the Mini International Neuropsychiatric Interview (MINI) Suicide module, the Self-Rated Health Measurement Scale (SFHMS), the Self-Esteem Scale (SES), the Simplified Coping Style Questionnaire (SCQ), and the Perceived Stress Scale-10 (PSS-10). Over half of the students suffered from at least one mental health problem. Approximately 65.55% of freshmen had depression, and 46.85% had anxiety. Minority status, low family income, and religious belief were significantly associated with current mental health problems. These findings indicate that mental disorders are highly prevalent among the freshman student population. The prevalence of such mental disorders was greater than that of the general population, and the majority of students with mental health problems require treatment.

Effects of postnatal growth retardation on early neurodevelopment in premature infants with intrauterine growth retardation / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the effects of postnatal growth retardation on early neurodevelopment in premature infants with intrauterine growth retardation (IUGR).</p><p><b>METHODS</b>A retrospective analysis was performed on the clinical data of 171 premature infants who were born between May 2008 and May 2012 and were followed up until a corrected gestational age of 6 months. These infants were classified into two groups: IUGR group (n=40) and appropriate for gestational age (AGA) group (n=131). The growth retardation rates at the corrected gestational ages of 40 weeks, 3 months, and 6 months, as well as the neurodevelopmental outcome (evaluated by Gesell Developmental Scale) at corrected gestational ages of 3 and 6 months, were compared between the two groups.</p><p><b>RESULTS</b>The growth retardation rate in the IUGR group was significantly higher than in the AGA group at the corrected gestational ages of 40 weeks, 3 months, and 6 months. All five developmental quotients evaluated by Gesell Developmental Scale (gross motor, fine motor, language, adaptability and individuality) in the IUGR group were significantly lower than in the AGA group at the corrected gestational ages of 3 months. At the corrected gestational age of 6 months, the developmental quotients of fine motor and language in the IUGR group were significantly lower than in the AGA group, however, there were no significant differences in the developmental quotients of gross motor, adaptability and individuality between the two groups. All five developmental quotients in IUGR infants with catch-up lag in weight were significantly lower than in IUGR and AGA infants who had caught up well.</p><p><b>CONCLUSIONS</b>Growth retardation at early postnatal stages may adversely affect the early neurodevelopment in infants with IUGR.</p>

Effects of HIV-1 tat on secretion of TNF-α and IL-1β by U87 cells in AIDS patients with or without AIDS dementia complex / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To explore the role of HIV-1 tat gene variations in AIDS dementia complex (ADC) pathogenesis.</p><p><b>METHODS</b>HIV-1 tat genes derived from peripheral spleen and central basal ganglia of an AIDS patient with ADC and an AIDS patient without ADC were cloned for sequence analysis. HIV-1 tat gene sequence alignment was performed by using CLUSTAL W and the phylogentic analysis was conducted by using Neighbor-joining with MEGA4 software. All tat genes were used to construct recombinant retroviral expressing vector MSCV-IRES-GFP/tat. The MSCV-IRES-GFP/tat was cotransfected into 293T cells with pCMV-VSV-G and pUMVC vectors to assemble the recombinant retrovirus. After infection of gliomas U87 cells with equal amount of the recombinant retrovirus, TNF-α, and IL-1β concentrations in the supernatant of U87 cells were determined with ELISA.</p><p><b>RESULTS</b>HIV-1 tat genes derived from peripheral spleen and central basal ganglia of the AIDS patient with ADC and the other one without ADC exhibited genetic variations. Tat variations and amino acid mutation sites existed mainly at Tat protein core functional area (38-47aa). All Tat proteins could induce U87 cells to produce TNF-α and IL-1β, but the level of IL-1β production was different among Tat proteins derived from the ADC patient's spleen, basal ganglia, and the non-ADC patient's spleen. The level of Tat proteins derived from the ADC patient's spleen, basal ganglia, and the non-ADC patient's spleen were obviously higher than that from the non-ADC patient's basal ganglia.</p><p><b>CONCLUSION</b>Tat protein core functional area (38-47aa) may serve as the key area of enhancing the secretion of IL-1β. This may be related with the neurotoxicity of HIV-1 Tat.</p>

Antimicrobial susceptibility of Streptococcus sp. to quinupristin-dalfopristin in China / 生物医学与环境科学（英文）

This study aimed to determine the in vitro activity of quinupristin-alfopristin against Streptococcus sp. isolated in China. This agent is not yet available for clinical use, but it has been tested against a high proportion of resistant Staphylococcus aureus strains. A total of 156 streptococcal isolates, which were recovered from various geographic areas and diseases, were tested using the Etest (AB Biodisk, Solna, Sweden). Quinupristin-alfopristin showed excellent activity against all of the tested streptococci isolates. These results provide useful data for the clinical use of quinupristin-alfopristin in China.

Progress in research of new human enterovirus types / 病毒学报

More and more new human enteroviruses (HEVs) types were identified with the broad application of the molecular serotyping methods for enteroviruses. Since enterovirus 71 (EV71) was first reported in 1969, numerous epidemic outbreaks associated with new enteroviruses have occurred all around the world, and pose a significant threat to public health . The epidemics of hand, foot and mouth disease (HFMD) caused by EV71 infection in China have raised great concern of global scholars. This paper reviewed research progress in recent years of the molecular typing, evolution, epidemiology, and pathogenesis attributable to new enterovirus types.

Molecular epidemiological characteristics of Streptococcus pyogenes strains involved in an outbreak of scarlet fever in China, 2011 / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To investigate molecular characterization of streptococcus pyogenes isolates involved in an outbreak of scarlet fever in China in 2011.</p><p><b>METHODS</b>Seventy-four Streptococcal pyogenes involved in an outbreak of scarlet fever were isolated from pediatric patients in the areas with high incidence in China from May to August of 2011. Emm genotyping, pulsed-field gel electrophoresis (PFGE), superantigen (SAg) genes and antimicrobial susceptibility profiling were analyzed for these isolates.</p><p><b>RESULTS</b>A total of 4 different emm types were identified. Emm12 was the most prevalent type which contained four predominating PFGE patterns corresponding to four different virulence and superantigen profiles. Emm12 (79.7%) and emm1 (14.9%) accounted for approximately 94% of all the isolates. The speA gene was all negative in emm12 isolates and positive in emm1 isolates. All strains were resistant to erythromycin, and 89.4% of them were resistant to erythromycin, tracycline, and clindamycin simultaneously.</p><p><b>CONCLUSION</b>Several highly diversified clones with a high macrolide resistance rate comprise a predominant proportion of circulating strains, though no new emm type was found in this outbreak. The data provide a baseline for further surveillance of scarlet fever, which may contribute to the explanation of the outbreak and development of a GAS vaccine in China.</p>

Analysis of genetic diversity of HIV-1 nef genes from a patient with AIDS dementia complex / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study the genetic diversity of HIV-1 nef genes from a patient with AIDS dementia complex(ADC) , so as to research the amino acid variability and the pathogenesis of ADC.</p><p><b>METHODS</b>The nef gene was amplified with PCR from genomic DNA which was extracted from spleen and different brain tissues(basal ganglia, frontal gray matter, meninges, temporal lobe)of a patient who died of ADC. PCR products were cloned into the pMD19-T vector, after transformation and selection by ampicillin and blue/white spotting. Five of positive clones were sequenced and confirmed with BLAST. HIV-1 nef sequences were processed with BioEdit and MEGA4 to do Neighbor-Joining tree, p-Distances, and values of ds/dn.</p><p><b>RESULTS</b>The samples were all identified as HIV-1 B and genetic variation exists in HIV-1 nef gene isolated from different tissues compared with HXB2. In addition,part of the changes were different between periphery and brain.</p><p><b>CONCLUSION</b>Variations exist in the HIV-1 nef gene extracted from the ADC patient and the variations from peripheral and central nerve tissues were different,these variations may change the function of Nef,and it needs more research.</p>

Expression and bioassay of rubella virus E1-374 glycoprotein in yeast cells / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To express the rubella virus E1-374 glycoprotein in Pichia pastoris and study the immunogenecity of the recombinant protein.</p><p><b>METHODS</b>The cDNA of protein E1-374 was cloned into the expression vector pGAPZalphaA and transformed into Pichia pastoris GS115 cells by electrotransfection. The expressed protein was confirmed by indirect immunofluorescence and demonstrated immunoreactivity by Western Blot. Rubella virus IgG antibody was assayed with ELISA after mice were inmmunized by E1-374 glycoprotein.</p><p><b>RESULTS</b>SDS-PAGE analysis and Western Blot analysis of E1-374 protein revealed this protein to be 46.89 x 10(3). Antiserum (1:100) and E1-374 (5.5 microg/ml) was chosen for ELISA optimization. The intra-assay coefficient of variation for the ELISA was 0.36%-12.45%.</p><p><b>CONCLUSION</b>Protein E1-374 was highly expressed in Pichia pastoris cells, and it was a good choice to prepare rubella virus recombinant protein vaccines.</p>

Effects of recombinant human erythropoietin on neurointelligence development in very low birth weight infants / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To evaluate the clinical effects of the early use of recombinant human erythropoietin (rhEPO) on the neurointelligence development in very low birth weight infants (VLBWI).</p><p><b>METHODS</b>Seventy-eight VLBWI were divided into rhEPO treatment group (n=35) and control group (n=43) according to the choice of their parents. Neonatal behavioral neurological assessment (NBNA) was performed at 40 weeks of corrected gestational age. The Gesell Developmental Schedules were used for neurodevelopmental evaluation at 3, 6, and 12 months of corrected age. The abnormal rates of auditory brainstem response (ABR) and cranial ultrasound were evaluated at 6 months of corrected age.</p><p><b>RESULTS</b>The rhEPO treatment group had significantly higher NBNA scores at 40 weeks of corrected gestational age than the control group (P<0.05). The adaptability at 3 months of corrected age, the gross motor, adaptability, and sociability at 6 months, and the gross motor, adaptability, fine motor, sociability, and language at 12 months were significantly better in the rhEPO treatment group than in the control group (P<0.05). The abnormal rates of ABR and cranial ultrasound in the rhEPO treatment group were significantly lower than in the control group at 6 months of corrected age (P<0.05).</p><p><b>CONCLUSIONS</b>Early use of rhEPO can promote the early recovery of neurological symptoms and improve the cognitive, motor, and language abilities in VLBWI due to its protective effects on the nervous system.</p>

Study on functions of N-carbohydrate chains in human parainfluenza virus type 3 hemagglutinin-neuraminidase protein / 病毒学报

To determine the functions of N-carbohydrate chains in human parainfluenza virus type 3 hemagglutinin-neuraminidase(HN) protein, a PCR-based site-directed mutagenesis method was used to obtain N-glycan mutants. Protein electrophoresis rate, cell surface expression,receptor binding activity, neuraminidase activity and cell fusion promotion activity were determined. The HN proteins of single mutants (G1, G2, and G4) and multiple mutants (G12, G14, G24 and G124) migrated faster than the wild-type (wt) HN protein on polyacrylamide gels, while G3-mutated protein and wt HN protein migrated at the same position. There was no statistic difference in cell surface expression and neuraminidase activity between wt and each mutant HN protein (P>0.05), but receptor binding activity and cell fusion promotion activity of each mutant protein was reduced to significant extent (P<0.05). G1, G2 and G4 mutants exhibited re duced receptor binding activity, which was 83.94%, 76.45% and 55.32% of the wt level, respectively. G1, G2 and G4-mutated proteins also showed reductions in fusion promotion activity, which was 80.84%, 77.83% and 64.16%, respectively. Multiple mutants with G12-, G14-, G24- and G124- substitutions could further reduce receptor binding activities, 33.07%, 20.67%, 19.96% and 15.11% of the wt HN level, respectively. G12, G14, G24 and G124 mutants exhibited levels of fusion promotion activity that were only 46.360, 12.04%, 13.43% and 4.05% of the wt amount, respectively. As N-glycans of hPIV3 HN protein play an important role in receptor binding activity and cell fusion promotion activity of HN protein. We propose that the loss of N-glycans change the conformation or orientation of globular domain that is responsible for receptor binding and lower receptor binding activity and cell fusion promotion activi ty.

Cloning and eukaryotic expression of HIV-1B gp120 genes from a patient with AIDS dementia complex / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To clone and express the HIV-1B gp120 genes isolated at different organizations from a patient died of AIDS dementia complex (ADC) in eukaryotic cells.</p><p><b>METHODS</b>Using the genomic DNA isolated from peripheral lymphnodes, choroid plexus and occipital white matter from a patient died of ADC as the template, HIV-1B gp120 gene was amplified with PCR. After sequenced, HIV-1B gp120 was inserted into pcDNA3.1 (+) and recombinant expressing vector gp120/pcDNA3.1 (+) was constructed succeffuly confirming with sequencing. Then expressing vector was transfected into eukaryotic cells U87 using liposome transfection and expression of HIV-1B gp120 gene was assayed with indirect immunofluorescence.</p><p><b>RESULTS</b>HIV-1B gp120 genes isolated from peripheral lymphnodes, choroid plexus and occipital white matter of the ADC patient were successfully cloned and recombinant expressing vector gp120/pcDNA3; 1 (+) could express envelope glycoprotein HIV-1B gp120 in U87 cells.</p><p><b>CONCLUSION</b>All the HIV-1B gp120 gene isolated at the different organizations of the same ADC patient could express in U87 cells, which may supply a valuable basis for studying the neurotoxicity and neurotoxic mechanism of HIV-1 gp120 protein.</p>

Analysis of genetic diversity and amino acid sequence of HIV-1tat in CNS and peripheral tissue of a patient with ADC and a patient with non-ADC / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study the diversity of HIV-1 tat gene in CNS and peripheral tissue of a patient with ADC and a patient with non-ADC, so as to research HIV evolution, the mechanism of CNS invasion and the pathogenesis of ADC.</p><p><b>METHODS</b>The tat gene was amplified with nested PCR from genomic DNA which was extracted from spleen and basal ganglia of one non-ADC patient with a wide range of cerebral artery atherosclerosis and one ADC patient. PCR products were cloned into the PGEM-T vector, after transformation and selection by ampicillin and blue/white spotting. Five of positive clones were sequenced. HIV-1 tat sequences were processed with BioEdit and MEGA4. With the softwares, neighbor-joining tree, p-distances, values of ds/dn, and analysis of amino acid motifs were all done, so as to research the diversity of HIV-1 tat gene in CNS and peripheral tissue.</p><p><b>RESULTS</b>Gene mutation of HIV-1 tat exist in the two patients, the mutation process of tat isolated from ADC patient suffered more compartmentalization than tat isolated from non-ADC patient, the differences of tat genes between CNS and peripheral tissue in ADC patient were greater than the non-ADC patient. Ds/dn showed that the virus gene mutation played a major role, the body intend to remove harmful non-synonymous mutations.</p><p><b>CONCLUSIONS</b>The compartmentation of tat gene in CNS and peripheral tissue of the two patients was different, the reason may be related to the pathway of HIV into the CNS, the relationship between HIV gene mutation in CNS and ADC still need more investigation.</p>