Investigation on seroprevalence of Toxoplasma gondii infections among neonates in Fujian Province / 中国血吸虫病防治杂志

Objective To investigate the seroprevalence of Toxoplasma gondii infections among neonates in Fujian Province, so as to provide insights into the development of interventions for the prevention and control of congenital toxoplasmosis. Methods A total of 1 045 neonates delivered in Fujian Province from 2017 to 2018 were recruited, including 387 preterm infants and 658 full-term infants. Umbilical cord blood was sampled from all neonates, and the seroprevalence of anti-T. gondii IgG antibody was detected and compared between preterm and full-term infants. In addition, elbow venous blood samples were collected from neonates’mothers, and the seroprevalence of anti-T. gondii IgG antibody was detected and compared between preterm and full-term infants’mothers. Results The overall seroprevalence of anti-T. gondii IgG antibody was 9.38% among the 1 045 neonates in Fujian Province. The seroprevalence of anti-T. gondii IgG antibody was 18.35% in the 387 preterm infants, and there was no significant difference in the seroprevalence of anti-T. gondii IgG antibody between male and female infants (17.69% vs. 18.75%, χ2 = 0.07, P > 0.05). The seroprevalence of anti-T. gondii IgG antibody was 4.10% in the 658 full-term infants, and there was no significant difference in the seroprevalence of anti-T. gondii IgG antibody between male and female infants (4.14% vs. 4.08%, χ2 = 0, P > 0.05). In addition, the overall seroprevalence of anti-T. gondii IgG antibody was 15.02% in all neonates’ mothers, and the seroprevalence was significantly greater in preterm infants’mothers than in full-term infant’s mothers (20.93% vs. 11.55%, χ2 = 16.79, P < 0.01). Conclusions The seroprevalence of T. gondii infections is significantly higher in preterm infants and their mothers than in full-term infants and their mothers. Prenatal detection of T. gondii infections and health education pertaining to toxoplasmosis prevention and control knowledge are required to be strengthened to effectively reduce the incidence of congenital toxoplasmosis.

Preliminary observation and analysis of the characteristics of gut microbiota in patients with schizophrenia / 中国神经精神疾病杂志

Objective To investigate the characteristics of gut microbiota in patients with schizophrenia. Methods The composition of the fecal microbiota in 16 schizophrenia patients and 20 healthy controls was analyzed by 16S rRNA sequencing. Results At the phylum level, TM7 was increased in the patients (P<0.05). At the class level, the abundance of Peptostreptococcaceae, Lactobacillaceae and Actinomycetaceae in the patients increased (P<0.05), whereas the abundance of Tissierellaceae decreased relative to health controls (P<0.05). At the genus level, Actinomyces, Scardovia, Atopobium, Moryella, G07 and Lactobacillus were increased in the patients relative to healthy controls (P<0.05). Prediction of functional properties of bacterial flora showed that the metabolic pathways of fatty acids and pyruvate were different between the patients and healthy controls (P<0.05). Conclusion The present study reveals an alteration in the characteristics of gut microbiota in schizophrenia patients which may be an auxiliary diagnosis marker for clinical diagnosis of schizophrenia.

Berberine enhances mitomycin C-induced cell cycle arrest and apoptosis of T24 bladder cancer cells / 中国病理生理杂志

AIM:To observe the effects of the combination of berberin (Ber) and mitomycin C (MMC) on the cell cycle arrest and apoptosis of T24 bladder cancer cells and the underlying mechanisms. METHODS:The T24 cells were exposed to MMC in the presence or absence of difference concentrations of Ber. The viability of the T24 cells was de-termined by CCK-8 assay. The cell cycle distribution was detected by flow cytometry. The apoptosis was analyzed by flow cytometry with Annexin V-FITC/PI staining, and the protein expression levels of cyclin D1, survivin, CDK2, CDK4, p21 and p27 were determined by Western blot. RESULTS:CCK-8 experiments showed that Ber enhanced the inhibitory effect of MMC on the viability of T24 cells. The results of flow cytometry showed that Ber also enhanced the blockade effect of MMC on T24 cells in G0/G1 phase (P<0.05). Compared with the MMC group, Ber increased the expression of p21 and p27 up-regulated by MMC, and decreased the expression of cynlin D1, CDK2 and CDK4 (P<0. 05). Meanwhile, Ber promoted MMC to inhibit the expression of survivin (P<0. 05). Ber increased the apoptosis of T24 cells induced by MMC (P<0. 05). CONCLUSION:Ber significantly enhances the inhibitory effect of MMC on the viability of T24 cells. The mechanism may be related to up-regulation of p21 and p27, thereby inhibiting the expression of cyclin D1, CDK-2 and CDK-4. At the same time, Ber inhibits the protein expression of survivin, which eventually leads to cell arrest in G0/G1 phase and promotes apoptosis.

Inhibition of anterior cingulate cortex excitatory neuronal activity induces conditioned place preference in a mouse model of chronic inflammatory pain

The anterior cingulate cortex (ACC) is known for its role in perception of nociceptive signals and the associated emotional responses. Recent optogenetic studies, involving modulation of neuronal activity in the ACC, show that the ACC can modulate mechanical hyperalgesia. In the present study, we used optogenetic techniques to selectively modulate excitatory pyramidal neurons and inhibitory interneurons in the ACC in a model of chronic inflammatory pain to assess their motivational effect in the conditioned place preference (CPP) test. Selective inhibition of pyramidal neurons induced preference during the CPP test, while activation of parvalbumin (PV)-specific neurons did not. Moreover, chemogenetic inhibition of the excitatory pyramidal neurons alleviated mechanical hyperalgesia, consistent with our previous result. Our results provide evidence for the analgesic effect of inhibition of ACC excitatory pyramidal neurons and a prospective treatment for chronic pain.

An Open-label, Self-control, Prospective Study on Cognitive Function, Academic Performance, and Tolerability of Osmotic-release Oral System Methylphenidate in Children with Attention-deficit Hyperactivity Disorder / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Attention-deficit hyperactivity disorder (ADHD) is the most common mental and behavioral disorder in school-aged children. This study evaluated the effect of osmotic-release oral system (OROS) methylphenidate (MPH) on cognitive function and academic performance of Chinese school-aged children with ADHD.</p><p><b>METHODS</b>This 12-week, prospective, multicenter, open-label, self-controlled study enrolled 153 Chinese school-aged children with ADHD and 41 non-ADHD children. Children with ADHD were treated with once-daily OROS-MPH (18 mg, 36 mg, or 54 mg). The primary endpoints were Inattention/Overactivity (I/O) with Aggression Conners Behavior Rating Scale (IOWA) and Digit Span Test at week 12 compared with baseline. Secondary endpoints included opposition/defiant (O/D) subscale of IOWA, Clinical Global Impression (CGI), Coding Test, Stroop Color-word Test, Wisconsin Card Sorting Test (WCST), academic performance on teacher-rated school examinations, and safety at week 12 compared with baseline. Both non-ADHD and ADHD children received the same frequency of cognitive operational test to avoid the possible bias caused by training.</p><p><b>RESULTS</b>A total of 128 patients were evaluated with cognitive assessments. The OROS-MPH treatment significantly improved IOWA Conners I/O subscale scores at week 12 (3.8 ± 2.3) versus baseline (10.0 ± 2.4; P < 0.0001). Digit Span Test scores improved significantly (P < 0.0001) with a high remission rate (81.1%) at week 12 versus baseline. A significant (P < 0.0001) improvement was observed in O/D subscale of IOWA, CGI, Coding Test, Stroop Color-word Test, WCST, and academic performance at week 12 versus baseline. Very few practice-related improvements were noticed in the non-ADHD group at week 12 compared with baseline. No serious adverse events and deaths were reported during the study.</p><p><b>CONCLUSIONS</b>The OROS-MPH treatment effectively controlled symptoms of ADHD and significantly improved academic performance and cognitive function of Chinese school-aged children with ADHD. The treatment was found to be safe and generally well-tolerated over 12 weeks.</p><p><b>TRIAL REGISTRATION</b>ClinicalTrials.gov, NCT01933880; http://clinicaltrials.gov/ct2/show/NCT01933880?term=CONCERTAATT4099&rank=1.</p>

Prokaryotic soluble expression of protein D of Haemophilus influenzae type b / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To express the recombinant D protein in prokaryotic expression system solubly and make preparation for producing D-carrier conjugate vaccine next step.</p><p><b>METHODS</b>The hpd gene fragment removed of signal peptide from genomic DNA of Hib CMCC was inserted into pET43. 1a. The recombinant plasmid was transformed to competent E. coli BL21 (DE3) for expression under induction of IPTG. The expressed recombination protein was precipitated with ammonium sulfate, purified by DEAE anion exchange column chromatography and identified for reactogenicity by Western Blot.</p><p><b>RESULTS</b>The expressed recombination protein, in a soluble form, constained about 50% of total somatic protein and showed specific reaction with the HIB antisera after preliminary purification.</p><p><b>CONCLUSION</b>The D protein recombined expression plasmid was constructed successfully and expressed D protein in prokaryotic cells in a solube form.</p>

Construction of recombinant CHO cell strain for high expression of HBsAg / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To overexpress hepatitis B virus S gene in CHO cells cultured in serum-free media.</p><p><b>METHOD</b>Plasmid was constructed by cloning of HBV S gene and then it was transfected into CHO cells. After cell screen, the positive clones were identified and isolated into a serum-free media followed by the serological and morphological characterization of the expression product.</p><p><b>RESULT</b>CHO cell strains which can express HBsAg efficiently and stably were obtained. Spherical and filamentous HBsAg could be detected under electronic microscope. The titer of the expression product was up to 1:5000.</p><p><b>CONCLUSION</b>Serum-free media cultured CHO cell strain for overexpression of HBsAg was successfully constructed and the expression product was high antigenic.</p>

Antigenic analysis of two chimeric hepatitis B core particles presenting the preS1 neutralizing epitopes / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To construct full-length hepatitis B core particles presenting preS1 aa 21-47 epitope and truncated core particles presenting preS1 aa 37-45 epitope on their surface and compare their antigenicity.</p><p><b>METHODS</b>PreS1 aa21-47 epitope and aa 37-45 epitope were inserted respectively into full-length hepatitis B core (aa 1-183) and truncated HBcAg (aa 1-144), between the 78th (Asp) and 79th (Pro). The genes synthesized after the codon optimization were ligated to the pET43. 1a vector with the same cohesive terminal (NdeI and XhoI) and expressed in the E. coli expression system. The morphology of the proteins of interest were observed by electron microscope and characterized by ELISA and Western Blotting.</p><p><b>RESULTS</b>The morphology of the virus-like particles were confirmed by electron microscope. H2 were solid particles with a diameter of (31.61 +/- 1.27) nm, while H3 were hollow particles with a diameter of (28.46 +/- 1.16) nm. Statistical analysis showed that H2 is larger than H3 in the diameter (P < 0.01). The antigenicity of the inserted epitopes and carrier protein were identified by ELISA and Western Blotting.</p><p><b>CONCLUSION</b>Chimeric hepatitis B core particles presenting the preS1 neutralizing epitopes on their surface have been expressed, purified and identified, which lays the foundation for its application in vaccine research.</p>

Effect of gene optimization on the expression and purification of HDV small antigen produced by genetic engineering / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study the effect of gene optimization on the expression and purification of HDV small antigen produced by genetic engineering.</p><p><b>METHODS</b>Based on the colon preference of E. coli, the HDV small antigen original gene from GenBank was optimized. Both the original gene and the optimized gene expressed in prokaryotic cells, SDS-PAGE was made to analyze the protein expression yield and to decide which protein expression style was more proportion than the other. Furthermore, two antigens were purified by chromatography in order to compare the purity by SDS-PAGE and Image Lab software.</p><p><b>RESULTS</b>SDS-PAGE indicated that the molecular weight of target proteins from two groups were the same as we expected. Gene optimization resulted in the higher yield and it could make the product more soluble. After chromatography, the purity of target protein from optimized gene was up to 96.3%, obviously purer than that from original gene.</p><p><b>CONCLUSION</b>Gene optimization could increase the protein expression yield and solubility of genetic engineering HDV small antigen. In addition, the product from the optimized gene group was easier to be purified for diagnosis usage.</p>

Detection of hepatitis A virus RNA with an improved loop-mediated isothermal amplification assay / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To establish a novel improved loop-mediated isothermal amplification (LAMP) technique to detect hepatitis A virus (HAV).</p><p><b>METHODS</b>A novel improved LAMP assay based on the addition of an acceleration primer was developed for hepatitis A virus nucleotide acid detection.</p><p><b>RESULTS</b>Precision and reproducibility analysis proved its high stability and reliability. Comparison between the improved and conventional LAMP assays revealed that the former was more sensitive with a detection limit of 5 TCID50/ml. The novel detection method displayed 100% consistency with the TaqMan real-time PCR assay when applied to clinical specimens collected from patients with confirmed acute HAV infection.</p><p><b>CONCLUSION</b>This novel technique is widely applicable as a simple diagnostic tool in the clinical field as well as for the surveillance and investigation of the infectious disease in developing countries where HAV is endemic.</p>

Gene optimization, protein expression, purification and characterization of the HDV antigen for diagnosis / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To prepare HDAg with biological activities as a candidate of diagnostic reagent.</p><p><b>METHODS</b>To synthesize HDV gene fragment after codon optimization. To construct a thio-fused recombinant plasmid based on M48 expression vector. To express in E. coli induced by IPTG. To purify the protein by affinity chromatography followed by characterization in ELISA:</p><p><b>RESULTS</b>Plasmid construction was verified by enzyme digestion. SDS-PAGE indicated the molecular weight of the protein was the same as we expectation. ELISA proved its affinity with HDV antibodies.</p><p><b>CONCLUSION</b>HDAg was obtained successfully and it will pave the road to the research of HDV diagnostic reagent.</p>

Cortical Depression and Potentiation: Basic Mechanisms for Phantom Pain

People experience the feeling of the missing body part long after it has been removed after amputation are known as phantom limb sensations. These sensations can be painful, sometimes becoming chronic and lasting for several years (or called phantom pain). Medical treatment for these individuals is limited. Recent neurobiological investigations of brain plasticity after amputation have revealed new insights into the changes in the brain that may cause phantom limb sensations and phantom pain. In this article, I review recent progresses of the cortical plasticity in the anterior cingulate cortex (ACC), a critical cortical area for pain sensation, and explore how they are related to abnormal sensory sensations such as phantom pain. An understanding of these alterations may guide future research into medical treatment for these disorders.

Correlation between renal artery resistance index and serum creatinine level early after renal transplantation / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the relationship between the resistance index (RI) of the renal artery and serum creatinine (Cr) level in patients early (within one month) after renal transplantation.</p><p><b>METHODS</b>A total of 123 patients receiving renal transplantation underwent examinations by color Doppler ultrasound for measurement of the RI of the renal artery within one month after the operation. According to the results of RI measurement, the patients were divided into RI≥0.75 and RI<0.75 groups for analyzing the correlation between RI and serum Cr level measured at the same time points.</p><p><b>RESULTS</b>The RI and Cr levels in patients with RI≥0.75 showed a significant positive correlation (P<0.05), whereas they showed an inverse correlation in patients with RI<0.75 (P<0.05). The patients with RI≥0.75 had significantly lower RI of the renal artery and Cr level than those with RI≥0.75.</p><p><b>CONCLUSION</b>RI is significantly correlated to Cr, and may serve as an indicator for predicting renal graft function after transplantation.</p>

Construction and characterization of hepatitis B surface antigen "a" epitope virus-like particles / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To construct virus-like particles of hepatitis B core antigen, with HBsAg "a" epitope exposed on the surface.</p><p><b>METHODS</b>Hepatitis B surface antigen "a" epitope were inserted into the Hepatitis B core antigen, between the 78th (Asp) and the 79th (Pro) amino acids. The gene was synthesized after the codon optimized, then it was ligated to the express vector after been enzyme digest. The virus-like particles were observed by electron microscope and detected by ELISA after been expressed and purified. Immune the rabbits by the VLPs, then detect the antibody.</p><p><b>RESULT</b>The virus-like particles were confirmed by electron microscope. Its antigenicity and immunogenicity were identified by ELISA.</p><p><b>CONCLUSION</b>The prokaryotic express plasmid with the fusion gene has been constructed successfully. The virus-like particles have been expressed, purified and identified, which lays the foundation for its application in the further.</p>

Expression of hepatitis C virus subunit fusion protein and analysis of its immunogenicity / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>Obtain the hepatitis C virus high purified subunit fusion protein and detect its immunogenicity.</p><p><b>METHODS</b>With the vector of pET-11d, fusion protein was expressed in Escherichia coli BL21 (DE3) after induced by IPTG. The protein was then purified by DEAE negative ion exchange chromatography and Ni2+ affinity chromatography. Western Blot analysis was used to detect the antigenicity of the fusion protein. At the same time, the sera were collected and prepared from the immunized experimental animals in order to investigate the immunogenicity of the protein by EIA.</p><p><b>RESULTS</b>High purified hepatitis C virus subunit fusion protein was obtained successfully. The EIA indicated that the fusion protein could elicit specific antibodies in the animals with very high titers.</p><p><b>CONCLUSION</b>The hepatitis C virus subunit fusion protein expressed in prokaryotic system was proved to have strong immunogenicity. It could provide some helpful and useful information to the hepatitis C virus prophylactic and therapeutic vaccine development.</p>

Preparation and identification of the monoclonal antibodies against VP1 capsid protein of Enterovius 71 / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To prepare monoclonal antibodies (McAbs) against VP1 capsid protein of Enterovirus 71.</p><p><b>METHODS</b>Two peptides, SP55 and SP70, containing amino acid 163-177 and 208-222 of VP1, were synthesized respectively. Immunized BALB/c mice with the synthetic peptides to establish the hybridoma cell strains secreting specific McAb to VP1. After the specific McAbs were prepared, identified and analyzed the titer by indirect ELISA assay. The positive clones were selected and their neutralization titer were determined by neutralization test.</p><p><b>RESULTS</b>Two high titered anti-VP1 antibodies secreted by the hybridoma cells showed good neutralization reaction with enterovirus 71 on RD cells, and the neutralization titer were 1:8 and 1:16 respectively.</p><p><b>CONCLUSION</b>Two high titered anti-VP1 antibodies, with good neutralization activity, secreted by the hybridoma cells, which lays the foundation for further study.</p>

Expression of recombinant rubella virus E1 protein and initial application for detecting of antibody / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To apply recombinant rubella virus envelope protein-1 (E1) to detect human rubella virus IgG antibody.</p><p><b>METHODS</b>Rubella virus E1 protein was expressed in E. coli, purified E1 protein was used as the antigen for the detecting of anti rubella in human sera in the way of enzyme linked Immunosorbant assay (ELISA).</p><p><b>RESULTS</b>The antigenicity of the recombinant protein was checked by WHO rubella sera panel. We detected 200 sera samples, which came from Guangxi Guilin. 93% of these samples were positive.</p><p><b>CONCLUSION</b>The antigenicity of recombinant E1 is a satisfied candidate antigen for the detecting of human rubella virus antibody. The prevalence of anti rubella virus IgG in Guangxi is 93%. It is at the some level compared with other provinces in China.</p>

Cloning of PD-1 gene and its prokaryotic expression in Escherichia coli / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To clone human PD-1 gene, construct a prokaryotic expression plasmid and express in E. coli.</p><p><b>METHODS</b>The human PD-1 cDNA was cloned by RT-PCR from the total RNA, which was extracted from peripheral blood lymphocyte cell of the patient with chronic hepatitis B. Recombinant PD-1 protein was been expressed and purified after the prokaryotic expression plasmid had been constructed. It was identified by SDS-PAGE, DNA sequencing and amino acid sequencing.</p><p><b>RESULTS</b>The PD-1 gene was cloned and confirmed by DNA sequencing. The recombinant protein was expressed in E. coli. The purified protein was obtained, then been confirmed by amino acid sequencing.</p><p><b>CONCLUSION</b>The human PD-1 gene was successfully cloned and expressed in E. coli, which lays the foundation for further study on the function and application of PD-1.</p>

Antigenic properties of mutant hepatitis B virus surface antigen / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study the antigenic properties of mutant hepatitis B virus surface antigen, to understand the sensitivity of the commercially available HBsAg assays to the variants and to reduce the undetectability of the variants.</p><p><b>METHODS</b>Recombinant eukaryotic expression plasmids for HBsAg. The recombinant eukaryotic expression plasmids pSS1adr, pSS1adw2, pSS1adw2- 145Arg, pSS1adr-126 Asn and pSS1adr-126Ser were transfected into COS-7 cells. HBsAg in the supernatants of transfected cells was detected by using different commercial ELISA kits.</p><p><b>RESULTS</b>The absorbance value of pSS1adr-126 Asn and pSS1adr-126Ser plasmids were similar to that of the wild type HBsAg, the absorbance value of pSS1adw2-145Arg plasmids was lower than that of the wild type HBsAg.</p><p><b>CONCLUSION</b>It is estimated that the antigenicity of HBsAg mainly depended on the amino acid sequence of "a" antigen determinant and its conformation, so 145 amino acid substitutions led to the change of conformation and the antigenicity of variant HBsAg was lower than that of the wild type.</p>

Expression of human IL-12 in mammalian cell and study on its biological activities / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To obtain recombinant human interleukin 12 by genetic engineering, and to explore possibility of its clinical application in treatment of tumor and chronic hepatitis.</p><p><b>METHODS</b>Bicistronic expression vector P35-IRES-P40 was constructed for the simultaneous translation of IL-12 p35 and p40 cDNA subunit through internal ribosomal entry sites (IRES). pCI-dhfr-P35-IRES-P40 vector was constructed for expression in CHO-DHFR- cells. Positively cloned cells were screened by means of ELISA. Pools of clones with increased expression of IL-12 could be generated by selection in methotrexate. To determine the biological activities of rhIL-12, PHA-activated lymphoblasts proliferation assay and IFN-gamma induction assay were used in this study.</p><p><b>RESULTS</b>Genetically engineered cells expressing hIl-12 were obtained and all the cell lines showed the stabile expression of rhIL-12 in high efficiency and good growth properties.</p><p><b>CONCLUSION</b>rhIL-12 have good biological activities, it can stimulate activation and proliferation of T cells and induce production of IFN-gamma.</p>