Efficacy evaluation and neurobiological mechanism of psychedelics based on looming model / 中国药理学与毒理学杂志

OBJECTIVE To investigate the effect of psychedelics on innate fear behavior of mice in Looming model(LM)and its neurobiological mechanism.METH-ODS ① Adult male C57BL/6J mice were randomly divid-ed into saline group,DOM group,psilocybin group and mescaline group.The drugs of the corresponding groups were given ip injecction 5 min in advance and LM were used to test the effect of them on freezing time in shelter of mice.② The mice were performing ip given DOM or psilocybin following 5-HT2A receptor antagonist M100907 ip 30 min later involved looming experiments.③Quan-tified the expression of EGR1 protein in mouse brains by immunofluorescence staining,then use ibotenic acid(IBO)damaged the mouse brain regions based on the result above and performed looming experiments.④ Specifically activate or inhibit CaMK Ⅱ,PV,VIP and SOM neurons of mice in saline or psilocybin groups respec-tively by chemical genetic methods and followed looming experiments.RESULTS ① In LM,the freezing time in shelter of mice in DOM,psilocybin and mescaline groups was significantly shorter compared to the saline group(P<0.05),and the dose-effect curves of above psyche-delics were U-shaped.② Compared with the vehicle + psilocybin/DOM groups,the freezing time in shelter of mice in M100907 + psilocybin/DOM groups was signifi-cantly prolonged(P<0.05),and there was no significant difference between the vehicle + saline group and the M100907 + psilocybin/DOM groups.③ Psilocybin signifi-cantly increased the expression of EGR1 protein in prelim-bic cortex(PrL)compared with saline,and the damage of PrL could effectively antagonized the effect of psilocybin shortening the freezing time in LM.④ Chemicalgenetic specific inhibition of CaMK Ⅱ,PV or VIP neurons in PrL could effectively antagonize the effect of psilocybin in LM,while chemicalgenetic specific activation of CaMK Ⅱneurons could significantly shorten the freezing time of saline-treated mice.CONCLUSION Psychedelics have capability to waken the innate fear behavior like freez-ing of mice in LM,and this effect is mediated by 5-HT2A receptor and CaMK Ⅱneuron in PrL.

Clinical and genetic characteristic in patients with disorders of sex development caused by Y chromosome copy number variant / 中华儿科杂志

Objective: To investigate the clinical phenotype and genetic characteristics of disorders of sex development (DSD) caused by Y chromosome copy number variant (CNV). Methods: A retrospective analysis was performed on 3 patients diagnosed with DSD caused by Y chromosome CNV admitted to the First Affiliated Hospital of Zhengzhou University from January, 2018 to September, 2022. Clinical data were collected. Clinical study and genetic test were performed by karyotyping, whole exome sequencing (WES), low coverage whole genome copy number variant sequencing (CNV-seq), fluorescence in situ hybridization (FISH) and gonadal biopsy. Results: The 3 children, aged 12, 9, 9 years, the social gender were all female, presented with short stature, gonadal dysplasia and normal female external genital. No other phenotypic abnormality was found except for case 1 with scoliosis. The karyotype of all cases were identified as 46, XY. No pathogenic vraiants were found by WES. CNV-seq determined that case 1 was 47, XYY,+Y(2.12) and case 2 was 46, XY,+Y(1.6). FISH concluded that the long arm of Y chromosome was broken and recombined near Yq11.2, and then produced a pseudodicentric chromosome idic(Y). The karyotype was reinterpreted as mos 47, X, idic(Y)(q11.23)×2(10)/46, X, idic(Y)(q11.23)(50) in case 1. The karyotype was redefined as 45, XO(6)/46, X, idic(Y)(q11.22)(23)/46, X, del(Y)(q11.22)(1) in case 2. 46, XY, -Y(mos) was found by CNV-seq in case 3, and the karyotype of 45, XO/46, XY was speculated. Conclusions: The clinical manifestations of children with DSD caused by Y chromosome CNV are short stature and gonadal dysgenesis. If there is an increase of Y chromosome CNV detected by CNV-seq, FISH is recommended to classify the structural variation of Y chromosome.

Detection of drug resistance genes of mycobacterium tuberculosis by rolling circle amplification technique with multicolor fluorescent probes / 中华预防医学杂志

The aim of this study was to construct a simple, rapid and ultra-sensitive optical biosensing technique based on rolling circle amplification (RCA), and to apply it to multiple detection of drug-resistant genes of mycobacterium tuberculosis. The common mutation sites of isoniazid, rifampicin and streptomycin resistance genes are katG315 (AGC➝ACC), rpoB531 (CAC➝TAC) and rpsL43 (AAG➝AGG). For these three gene sites, from February 2020 to May 2021, in the Department of Laboratory Medicine of the First Affiliated Hospital of Army Military Medical University, the padlock probe (PLP), primers and capture probes were designed. And a solid-phase RCA constant temperature amplification reaction system based on magnetic beads was constructed and the experimental parameters were optimized. The RCA products were accurately captured by the multicolor fluorescent probes (Cy3/Cy5/ROX), and the single-tube multiple detection of three mutation genes was realized. The sensitivity, specificity and linear range of this method were further verified. The results showed that the response range of katG315 in the same reaction system ranged from 1.0 pmol/L to 0.1 nmol/L. The response range of rpoB531 and rpsL43 ranged from 1.0 pmol/L to 50.0 pmol/L and 1.0 pmol/L to 20.0 pmol/L, and the method had good specificity and sensitivity, and could accurately identify single base mutations in mixed targets, with the minimum detection limit as low as 1.0 pmol/L. The recoveries of simulated serum samples were 95.0%-105.2%. In conclusion, the constant temperature amplification multiple detection method constructed in this study can quickly realize the single-tube multiple detection of three drug resistance mutation sites. This technology is low-cost, simple and rapid, and does not rely on large equipment, providing a new analysis method for pathogen drug resistance gene detection.

Effects of combined administration of loganin and berberine on bone metabolism in diabetic mice / 中国药理学通报

Aim To investigate the effects of combined administration of loganin and berberine on bone structure and metabolism in diabetic mice and its potential mechanism.Methods The diabetic ICR mouse model was induced by high fat diet(HFD).After 10 weeks of combined intervention, the effects of loganin and berberine on body weight, body fat rate, blood glucose, blood lipid and serum oxidative stress levels were observed.Bone microstructure was scanned by micro-CT.Biomechanical characteristics of bone were measured by three-point bending test, and material properties were detected by fourier transform infrared(FTIR).The pathological changes were observed by HE and TRAP staining.Protein expressions involved in advanced glycation end products(AGEs)and their receptors(RAGE)/nuclear factor-κB(NF-κB)signaling pathway were detected by immunohistochemistry.Results The combined administration of loganin and berberine could significantly inhibit the weight gain, reduce the levels of blood glucose, blood lipid and oxidative stress, as well as improve glucose tolerance.In addition, combined intervention also decreased the expression levels of the proteins involved in AGEs/RAGE/NF-κB signaling pathway, and improved bone microstructure, finally contributing to increasing bone quality in diabetic mice.Conclusions The combination of loganin and berberine could improve bone metabolism in diabetic mice, which may be related to AGEs/RAGE/NF-κB signaling pathway.

Molluscicidal effect of immersion with 50% wettable powder of niclosamide ethanolamine salt against Oncomelania hupensis on the soil surface and inside the soil layer in winter / 中国血吸虫病防治杂志

Objective To evaluate the molluscicidal effect of 50% wettable powder of niclosamide ethanolamine salt (WPNES) against Oncomelania hupensis on the soil surface and inside the soil layer by immersion method in winter. Methods O. hupensis snails were placed on the soil surface and 2, 5 cm and 10 cm under the soil layer outdoors in winter, and then immersed in 50% WPNES at concentrations of 1 mg/L and 2 mg/L for 1, 3 d and 7 d, while dechlorinated water served as controls. Snail mortality was observed following immersion with 50% WPNES on the soil surface and inside the soil layer. Results Following immersion with 50% WPNES at concentrations of 2 mg/L and 1 mg/L outdoors in winter, the 3-day corrected snail mortality rates were 98.0% and 76.0% on the soil surface, and the 7-day corrected snail mortality rate was both 100.0%. Following immersion with 50% WPNES at concentrations of 2 mg/L and 1 mg/L outdoors in winter, the 7-day corrected snail mortality rates were 95.5% and 85.6% 2 cm below the soil layer, 66.0% and 6.4% 5 cm below the soil layer. However, the 7-day snail mortality rate swere comparable between the 50% WPNES treatment group (at 2 mg/L and 1 mg/L) and controls 10 cm below the soil layer (both P > 0.05). Conclusion Immersion of 50% WPNES at a concentration of 2 mg/L for 7 days presents a high molluscicidal efficacy against O. hupensis on the soil surface and 5 cm within the soil layers in winter.

Association between glycated hemoglobin and ambulatory blood pressure or heart rate in hypertensive patients / 中南大学学报(医学版)

OBJECTIVES@#To determine the association between glycosylated hemoglobin (HbA1c) and ambulatory blood pressure or heart rate in hypertensive patients.@*METHODS@#A total of 585 patients, who performed ambulatory blood pressure monitoring (ABPM) from September 2018 to April 2019 in Xiangya Hospital, Central South University, were enrolled and assigned into 2 groups (470 in a hypertensive group and 115 in a normal group). HbA1c levels were compared. According to the HbA1c level, the hypertensive group was divided into 2 subgroups: A high HbA1c group (HbA1c≥6.1%, @*RESULTS@#The hypertensive group had higher HbA1c level than the normal group [(6.1±1.3)% vs (5.1±1.7)%, @*CONCLUSIONS@#In hypertensive patients, HbA1c is positively correlated with ambulate blood pressure, blood pressure load, and heart rate, and it has no correlation with blood pressure variability, heart rate variability, or morning blood pressure.

Cloning and enzymatic function characterization of a novel tropinone reductase I (DaTRI 2) in Datura arborea / 药学学报

Tropinone reductase I (TRI) is a key branch point enzyme in the midstream of tropane alkaloids (TAs) biosynthesis pathway and represents an important target for TAs metabolic engineering, which can lead to metabolic flux of substrate tropinone to TAs. A novel TRI gene was isolated from Datura arborea, a woody resource plant, and designated as DaTRI2 (GenBank accession number is MH705164). The full-length cDNA of DaTRI2 with 1 135 bp exhibits a high sequence homology (96.8%) with DaTRI, and is predicted to encode a protein of 347 amino acids. Deduced DaTRI2 protein contain a conserved TGXXXGXG motif involved in NADPH binding, the catalytic N-S-Y-K tetrad motif and eleven amino acid residues important for binding to its substrate tropinone. The phylogenetic analysis revealed that DaTRI2 and other TRIs from Solanaceous plants belong to the same cluster and DaTRI2 exhibited closest phylogenetic proximity to TRIs from Datura. DaTRI2 was expressed in E. coli and the purified recombinant protein can catalyze both tropinone reduction and tropine oxidation with an optimum pH value of 8.0 and 9.6, respectively. When tropinone was used as the substrate, the Km and Vmax values of DaTRI2 at pH 6.4 were 210.05 μmol·L-1 and 69.6 nkat·mg-1 protein respectively, while the Km and Vmax values for tropine as the substrate were 188.03 μmol·L-1 and 114 nkat·mg-1 protein respectively, at pH 9.6. DaTRI2 transcript was most abundant in the young leaf, followed by the root. Cloning of DaTRI2 gene and biochemical analysis of recombinant DaTRI2 facilitate further research on the molecular mechanism on TAs biosynthesis in woody plants and provide a more potent candidate for TAs metabolic engineering.

Seroepidemiological survey of hepatitis B virus infection among people under 60 years old in Ma’anshan City, 2017 / 中华疾病控制杂志

Objective To understand the current epidemic and immunity status of hepatitis B virus in Ma’anshan City, and to compare the prevention and control effect after the adjustment of hepatitis B vaccine immunization strategy. Methods Multi-stage random sampling method was used to select 10 investigation points in the whole city, a random sample of 3 460 people under 60 years old was included according to urban and rural stratification. questionnaires and blood were collected from the subjects, and domestic enzyme linked immunosorbent assay (ELISA) was used for hepatitis B immunoglobulins detection. Results The total positive rate of hepatitis B virus surface antigen (HBsAg), hepatitis B virus surface antibody (HBsAb) and Hepatitis B virus (HBV) infection was 3.32%, 51.21% and 29.16% respectively. The positive rate of HBsAb in urban area was higher than that in rural area ( 2=28.204, P<0.001). The positive rate of HBsAb was significantly different between the medical and nursing staff and other occupational groups ( 2=22.772, P<0.001). The difference of HBsAb positive rate before and after HepB vaccine content adjustment was statistically significant ( 2=90.331, P<0.001). The rate of HepB decreased with age ( 2trend=1 984.342, P<0.001). Conclusions Since HepB was incorporated into the immunization program, hepatitis B prevention and control in school-age children has achieved remarkable results. More attention should be paid on the low positive rate of HBsAb in students and the low immunization rate of HepB in adults.

Effects of constant light exposure on obesity in high fat diet rats / 中华内分泌代谢杂志

Objective To observe the effects of constant light exposure on the obesity in high fat diet rats. Methods Thirty-two male SD rats were randomly divided into four groups:rats on a normal chow exposed to standard light-dark cycle ( group A) , rats on a normal chow exposed to constant light ( group B) , rats on a high fat diet exposed to standard light-dark cycle ( group C) , and rats on a high fat diet exposed to constant light ( group D) . Body weights and food intakes were recorded weekly throughout the 12-week study. Body weight, fat mass, visceral adipose tissue weight, intraperitoneal glucose tolerance test ( IPGTT) results, insulin resistance parameters, serum lipids and levels of interleukin 6 (IL-6), tumor necrosis factor-α(TNF-α) were compared among groups. Epididymal adipose tissues mRNA expression of circadian clock genes, i. e. clock, bmal1, rorα, rev-erbα, cry1, per1, and per2 were analyzed by realtime PCR. Results From the 9th week, body weights of rats in group D were significantly higher than those in group C (all P<0. 05). At the 12th week, area under curve of IPGTT (AUC-IPGTT) in groups B, C, and D were significantly higher than that in group A. AUC-IPGTT in group D was significantly higher than that in group C (all P<0.05). Compared with group C,asignificant increase in fat mass,visceral adipose tissue weight,homeostasis model assessment for insulin resistance, serum cholesterol, TNF-α levels were observed in group D ( all P<0. 05). And a significant decrease in quantitative insulin sensitivity check index ( QUICKI) and high density lipoprotein-cholesterol were observed in group D in comparison with group C (both P<0. 05). Circadian clock genes (clock, rorα, rev-erbα, cry1, per1) mRNA expressions in group B and D were significantly different from those in group A (all P<0. 05) . Expression of cry1 in group D was significantly higher than that in group C. In group C, rev-erbαmRNA expression was significantly down-regulated in comparison with group A (P<0. 05). Conclusion Constant light exposure exaggerates obesity, glycolipid metabolism abnormality, inflammation, and insulin resistance in high fat diet rats.

Y-27632 reduces the MMP2 and MMP9 expression in endothelial cell via inhibition of ROCK signal pathway / 中南大学学报(医学版)

OBJECTIVE@#To explore the effect of ROCK inhibitor Y-27632 on the matrix metalloproteinase 2 and 9 (MMP2 and MMP9) gene expression and activity in tumor necrosis factor α (TNF-α)-treated human umbilical vein endothelial cell (HUVEC).
@*METHODS@#HHUVEC was divided into 3 groups, a control group, a TNF-α group, and a TNF-α plus Y-27632 group. The expressions of vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), MMP2 and MMP9 were examined by real-time PCR. The MMP2/9 activity was measured by gelatin zymography.
@*RESULTS@#Compared to the control group, the mRNA expressions of ICAM-1, VCAM-1, MMP2 and MMP9 were increased TNF-α-treated cells, which were suppressed by ROCK inhibitor (P<0.01). The MMP2/9 activity was elevated in TNF-α-treated cells, which was reversed by ROCK inhibitor (P<0.05).
@*CONCLUSION@#ROCK inhibitor can suppress TNF-α-induced inflammation in endothelial cells through down-regulation of MMP2/9.

Overexpression of NtPMT and HnH6H changed hyoscyamine-rich Atropa belladonna to scopolamine-rich varieties / 药学学报

Atropa belladonna L. is the commercial plant material for production of tropane alkaloids, including hyoscyamine and scopolamine. The wild-type Atropa belladonna is characterized by the hyoscyaminerich chemotype, in which the hyoscyamine content is much higher than the scopolamine content. It is the common goal for the pharmaceutical industry to increase the content of scopolamine in A. belladonna. Based on the T0 progeny of transgenic A. belladonna with NtPMT and HnH6H overexpression, T1 progeny of transgenic A. belladonna were obtained through self-pollination and used in a field trial. The 461 bp fragment of NtPMT and the 1 077 bp HnH6H were simultaneously expressed from T1 progeny of transgenic A. belladonna, but were not obtained from the wild-type A. belladonna. At the transcription level, the expression of NtPMT and HnH6H were detected in T1 progeny of transgenic A. belladonna, but were not detected in the wild-type plants. Further, the alkaloids were analyzed by HPLC. In the stems and leaves of T1 progeny of transgenic A. belladonna, hyoscyamine was not detected and scopolamine was detected at very high levels; in the stems and leaves of wild-type A. belladonna, hyoscyamine was detected at much higher levels. In the leaves of T1 progeny of transgenic A. belladonna, the content of scopolamine was 15-36 folds higher than that of wildtype leaves; in the stems of T1 progeny of transgenic A. belladonna, the scopolamine content was 37-108 folds higher than that of wild-type stems. In conclusion, overexpression of NtPMT and HnH6H greatly enhanced conversion of hyoscyamine into high-value scopolamine and improved the commercial value of A. belladonna.

Cloning and expression of the key enzyme hyoscyamine 6 beta-hydroxylase gene (DaH6H) in scopolamine biosynthesis of Datura arborea / 药学学报

Hyoscyamine 6 beta-hydroxylase (H6H) is the last rate-limiting enzyme directly catalyzing the formation of scopolamine in tropane alkaloids (TAs) biosynthesis pathway. It is the primary target gene in the genetic modification of TAs metabolic pathway. Full-length cDNA and gDNA sequences of a novel H6H gene were cloned from Datura arborea (DaH6H, GenBank accession numbers for cDNA and gDNA are KR006981 and KR006983, respectively). Nucleotide sequence analysis reveals an open reading frame of 1375 bp encoding 347 amino acids in the cDNA of DaH6H, while the gDNA of DaH6H contains four exons and three introns, with the highest similarity to the gDNA of H6H from D. stramonium. DaH6H also exhibited the most identity of 90.5% with DsH6H in amino acids and harbored conserved 2-oxoglutarate binding motif and two iron binding motifs. The expression level of DaH6H was highest in the mature leaf, followed by the secondary root, and with no expression in the primary root based on qPCR analysis. Its expression was inhibited by MeJA. DaH6H was expressed in E. coli and a 39 kD recombinant protein was detected in SDS-PAGE. Comparison of the contents of scopolamine and hyoscyamine in various TAs-producing plants revealed that D. arborea was one of the rare scopolamine predominant plants. Cloning of DaH6H gene will allow more research in the molecular regulatory mechanism of TAs biosynthesis in distinct plants and provide a new candidate gene for scopolamine metabolic engineering.

Cloning and expression of the key enzyme hyoscyamine 6 beta-hydroxylase gene (DaH6H) in scopolamine biosynthesis of Datura arborea / 药学学报

Hyoscyamine 6 beta-hydroxylase (H6H) is the last rate-limiting enzyme directly catalyzing the formation of scopolamine in tropane alkaloids (TAs) biosynthesis pathway. It is the primary target gene in the genetic modification of TAs metabolic pathway. Full-length cDNA and gDNA sequences of a novel H6H gene were cloned from Datura arborea (DaH6H, GenBank accession numbers for cDNA and gDNA are KR006981 and KR006983, respectively). Nucleotide sequence analysis reveals an open reading frame of 1375 bp encoding 347 amino acids in the cDNA of DaH6H, while the gDNA of DaH6H contains four exons and three introns, with the highest similarity to the gDNA of H6H from D. stramonium. DaH6H also exhibited the most identity of 90.5% with DsH6H in amino acids and harbored conserved 2-oxoglutarate binding motif and two iron binding motifs. The expression level of DaH6H was highest in the mature leaf, followed by the secondary root, and with no expression in the primary root based on qPCR analysis. Its expression was inhibited by MeJA. DaH6H was expressed in E. coli and a 39 kD recombinant protein was detected in SDS-PAGE. Comparison of the contents of scopolamine and hyoscyamine in various TAs-producing plants revealed that D. arborea was one of the rare scopolamine predominant plants. Cloning of DaH6H gene will allow more research in the molecular regulatory mechanism of TAs biosynthesis in distinct plants and provide a new candidate gene for scopolamine metabolic engineering.

Expression pattern of genes involved in tropane alkaloids biosynthesis and tropane alkaloids accumulation in Atropa belladonna / 中国中药杂志

Atropa belladonna is a medicinal plant and main commercial source of tropane alkaloids (TAs) including scopolamine and hyoscyamine, which are anticholine drugs widely used clinically. Based on the high throughput transcriptome sequencing results, the digital expression patterns of UniGenes representing 9 structural genes (ODC, ADC, AIH, CPA, SPDS, PMT, CYP80F1, H6H, TRII) involved in TAs biosynthesis were constructed, and simultaneously expression analysis of 4 released genes in NCBI (PMT, CYP80F1, H6H, TRII) for verification was performed using qPCR, as well as the TAs contents detection in 8 different tissues. Digital expression patterns results suggested that the 4 genes including ODC, ADC, AIH and CPA involved in the upstream pathway of TAs, and the 2 branch pathway genes including SPDS and TRII were found to be expressed in all the detected tissues with high expression level in secondary root. While the 3 TAs-pathway-specific genes including PMT, CYP80F1, H6H were only expressed in secondary roots and primary roots, mainly in secondary roots. The qPCR detection results of PMT, CYP80F1 and H6H were consistent with the digital expression patterns, but their expression levels in primary root were too low to be detected. The highest content of hyoscyamine was found in tender stems (3.364 mg x g(-1)), followed by tender leaves (1.526 mg x g(-1)), roots (1.598 mg x g(-1)), young fruits (1.271 mg x g(-1)) and fruit sepals (1.413 mg x g(-1)). The highest content of scopolamine was detected in fruit sepals (1.003 mg x g(-1)), then followed by tender stems (0.600 mg x g(-1)) and tender leaves (0.601 mg x g(-1)). Both old stems and old leaves had the lowest content of hyoscyamine and scopolamine. The gene expression profile and TAs accumulation indicated that TAs in Atropa belladonna were mainly biosynthesized in secondary root, and then transported and deposited in tender aerial parts. Screening Atropa belladonna secondary root transcriptome database will facilitate unveiling the unknown enzymatic reactions and the mechanisms of transcriptional control.

Correlation of serum calprotectin level with the range of coronary lesion in patients with acute coronary syndrome / 中南大学学报(医学版)

OBJECTIVE@#To examine the serum levels of S100 calcium-binding protein A8/A9 complex (S100A8/ A9) in patients with acute coronary syndrome (ACS) and to explore the relation between the serum levels of S100A8/A9 and the degree of coronary lesion.@*METHODS@#A total of 126 patients with coronary heart disease were enrolled from Xiangya Hospital of Central South University between September 2010 and January 2011, which included 51 patients with unstable angina pectoris (UAP group, n=51), 50 patients with acute myocardial infarction (AMI group, n=50), and 25 patients with stable angina pectoris (SAP group, n=25). Twenty-five healthy volunteers were served as a normal control group (NC group, n=25). According to the coronary artery lesion area, ACS patients were also divided into a single-branch group, a double-branch group and a triple-branch group. Serum levels of S100A8/A9 were measured by enzyme-linked immunosorbent assay on the day when the patients admitted to the hospital and on the day after one-week treatment (UAP group + AMI group). The serum levels were compared among the various branch groups. The short-term prognosis in patients with ACS was investigated by phone follow-up after 3 months.@*RESULTS@#1) The S100A8/A9 level in the SAP group was significantly higher than that in the normal control group (P0.05); 2) After one-week standard treatment, the serum levels of S100A8/A9 in patients with ACS were significantly reduced compared with that at the admission (P 0.05).@*CONCLUSION@#The serum level of S100A8/A9 is significantly elevated in patients with ACS, which might be positively correlated with the number of the coronary lesion branches.

Experimental study on polysaccharide of Cipangopaludina chinensis against HBV in vitro / 中国中药杂志

<p><b>OBJECTIVE</b>To evaluate the biological activity of polysaccharide of Cipangopaludina chinensis (PCC) against HBV in vitro.</p><p><b>METHOD</b>HepG2 2. 2. 15 cells were taken as the in vitro experimental model. The cell toxicity was observed by MTT. PCC of different safe concentrations and positive control medicine 3TC were added into the cells. Cell control without medicine was set at the same time. Cultural supernatants were collected at 9 d. HBsAg and HBeAg in cultural supernatants were tested by ELISA. The content of HBV-DNA was detected by TaqMan probe fluorescence quantitative PCR.</p><p><b>RESULT</b>TC0 and TC50 of PCC in HepG2 2. 2. 15 cell culture were 1 g . L-1 and >10 g . L-1, respectively, suggesting low toxicity in cells. IC50 of PCC in HepG2 2. 2. 15 cells HBsAg and HBeAg were 0. 501, 0. 401 g. L-1, with SI being >19.96 and >24. 94, respectively. PCC could effectively inhibit the secretion of HBsAg and HBeAg, and have a better effect on HBeAg than on HBsAg. PCC had a significant inhibitory effect on HBV-DNA in HepG2 2. 2. 15 cells at concentrations of 0. 1, 1 g . L-1 P <0.05).</p><p><b>CONCLUSION</b>PCC has the effect against HBV activity in vitro to some extent, with low toxicity, thereby having a good prospect for application.</p>

Development of a GeXP based multiplex RT-PCR assay for simultaneous detection of eight arboviruses related to encephalitis / 病毒学报

Multiplex reverse transcription-polymerase chain reaction (mRT-PCR) is currently available in virus detection and defined as the simultaneous amplification of two or more DNA/RNA targets in a single reaction vessel. In this study, we attempted to modify the conventional mRT-PCR technique on a basis of GenomeLab Genetic Analysis System (GeXP). Initially, we optimized the analytical validation of the GeXP analyzer and its design of workflow and simultaneously detected eight arboviruses that related to epidemic encephalitis by verifying the specificity of mRT-PCR with Japanese encephalitis virus(JEV) cell cultures and positive strains identified previously and determining the sensitivity with in vitro-transcribed RNA of serial dilutions. The GeXP system after optimization could amplify the specific fragments related to the viruses and exposed specifically a total of 13 target genes out of eight types of arboviruses at the level of 10(2) copies/microL, and the findings suggest that the novel protocol we developed can be high-throughput and highly specific and sensitive as well as quickness in screening of the encephalitis viruses, and is promising in detection of encephalitis-associated viruses for molecular epidemiological studies.

Human enterovirus 71 (EV71) resisted type I interferon induced antiviral effect / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate human enterovirus 71 (EV71) resistance to type I interferon induced antiviral effect.</p><p><b>METHODS</b>After type I interferons (alpha, beta) were incubated with HeLa cells, recombinant type I herpes simple virus (HSV-1) with green fluorescent protein (GFP) was inoculated onto the HeLa cells. HSV-1 proliferation was observed by GFP expression and PCR. After EV71 was inoculated onto HeLa cells incubated with the same quantity of interferon, proliferation of EV71 were detected by RT-PCR of 2A gene.</p><p><b>RESULTS</b>Recombinant HSV-1 GFP expression and viral DNA replication obviously decreased in HeLa cells incubated with type I interferon (alpha, beta). However, EV71 effectively proliferated in the interferon irritated HeLa cell by RT-PCR.</p><p><b>CONCLUSION</b>HeLa cell irritated by type I interferon (alpha, beta) produced antiviral substance that inhibits HSV-1 proliferation. EV71 resisted the antiviral substance induced by type I interferon and could significantly replicate in the HeLa cells.</p>

Influence of glucose concentration on the inhibition of Streptococcus oligofermentans on Streptococcus mutans / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the inhibition of Streptococcus oligofermentans (So) on Streptococcus mutans (Sm) and the producibility of hydrogen peroxide by So under the influence of glucose concentration environment.</p><p><b>METHODS</b>The inhibition between So and Sm was observed by plating method under the different glucose concentration environment. The initial synthesis rates and production of hydrogen peroxide by So were determined under the different glucose concentration environment by 4-aminoantipyine-horseradish peroxidase method at A(510).</p><p><b>RESULTS</b>Under 0, 10 and 50 mmol/L glucose environment, the inhibition of So on Sm was evident. When both Sm and So were inoculated at the same time, the ratio of inhibition area by bacterial membrane area was 0.202 ± 0.005, 0.467 ± 0.025, 0.468 ± 0.028 under 0, 10, 50 mmol/L glucose environment. When So was cultivated first and then Sm applied, the ratio was 0.394 ± 0.004, 0.811 ± 0.075 and 0.816 ± 0.007 under 0, 10 and 50 mmol/L glucose environment respectively. The inhibition under 10 and 50 mmol/L glucose environment were more significant than that under non-glucose environment. There was no significant difference between these two glucose concentrations (P > 0.05). The initial synthesis rates of H2O2 by So under the 10 mmol/L [(23.573 ± 0.263) µmo×L(-1)×min(-1)] and 50 mmol/L [(23.337 ± 0.473) µmol×L(-1)×min(-1)] glucose were higher than without glucose[(10.513 ± 0.516) µmol×L(-1)×min(-1)], P < 0.05. H2O2 was not detected in 1000 mmol/L glucose. However, the production of H2O2 by So under 0 mmol/L glucose was higher than other glucose concentrations (P < 0.05).</p><p><b>CONCLUSIONS</b>The capability of the inhibition of So on Sm was affected by glucose environment and was much stronger under certain glucose concentrations (10, 50 mmol/L).</p>

Plasma level of RBP4 in patients with coronary heart disease and the effect of hyperinsulinemia / 中南大学学报(医学版)

OBJECTIVE@#To evaluate the change of plasma level of retinol binding protein 4 (RBP4) in patients with coronary heart disease, and to explore the effect of hyperinsulinemia.@*METHODS@#This study was carried out at Xiangya Hospital of Central South University, China, from September 2009 to May 2010. Thirty patients with coronary artery disease (the CAD group) were confirmed by coronary angiography, 29 patients with CAD plus hyperinsulinemia (the CAD+HIns group), and 30 healthy subjects were enrolled as controls (the control group). The peripheral blood sample from the anticubital vein was collected aseptically in all the subjects to measure the RBP4 by enzyme linked immunosorbent-assay (ELISA). The height, weight, body mass index (BMI) the waist-to-hip ratio (WHR), the blood pressure, the fasting plasma glucose (FPG), the fasting insulin (Fins), the 2-hour postprandial inslulin (2hPIns), and the homeostasis model assessment-insulin resistance index (HOMA-IR) was measured. The lipids, high sensitivity C reactive protein (hsCRP), uric acid(UA), free fatty acids (FFA) were all examined.@*RESULTS@#The level of plasma RBP4 in the CAD+HIns group was higher than that in the CAD group and the control group (both P0.05). Correlation analysis showed that the plasma RBP4 level was significantly correlated with BMI, FPG, FIns, 2hPIns, HOMA-IR, TG, HDL-C, UA, and hsCRP (r=0.259, 0.331, 0.582, 0.452, 0.600, 0.236, -0.290, 0.243, 0.231, respectively; all P>0.05). Multiple regression analysis showed that BMI, 2hPIns, and HOMA-IR were the independent factors related to RBP4.@*CONCLUSION@#The plasma level of RBP4 does not increase in the CAD group, but it is high in the CAD +HIns group. RBP4 level is related to BMI, lipids, UA, and other cardiovascular risk factors. BMI, 2hPIns, and HOMA-IR are the independent factors associated with RBP4.