Study on the Effect of Lentivirus-Mediated NLRP3 Overexpression on Levels of Anti-CCP and RF in Synovial Fluid of Experimental RA Rabbits Regulated by Moxibustion / 世界科学技术-中医药现代化

Objective Observation of the anti-inflammatory and detumescence effects on experimental rheumatoid arthritis(RA)rabbits by moxibustion,With lentiviral vector mediated NLRP3 overexpression in RA animal models,to investigate the effect of moxibustion on anti-CCP/RF levels in synovial fluid of RA rabbit joints and its regulation mechanism of NLRP3.Methods Randomly divided thirty Japanese big-ear white rabbits into 5 groups:group of blank,group of model,group of moxibustion,NLRP3 overexpression group and NLRP3 overexpression negative group,with 6 rabbits in each group.Both joint of the model group,and the groups with moxibustion was injected into FCA(0.5 mL·kg-1),the blank group used the same method to inject sterile saline.On the 3rd day and the 10th day after treatment,10 μL of NLRP3 overexpression lentiviral vector was injected into the joint cavity of the NLRP3 overexpression group of rabbits.The rabbits in the NLRP3 overexpression negative group were injected with the same amount of LvGFP as Contrast.moxibustion bilateral"BL23","ST 36"of the groups with moxibustion(5 cones with one point in a day.6 days for 1 course,1 day for rest,3 courses of treatment in total.)After treatment,observe the circumference of each rabbit's left and right knee joints,the content of RF,anti-CCP,IL-18 in synovial fluid was detected by ELISA.Results Compared with the blank group,the left and right knee joint circumferences in the model group increased,and the anti-CCP,RF,and IL-18 contents in the synovial fluid of the knee joint increased significantly(P<0.01);compared with the modle group,The circumference of the knee joint decreased,and the content of anti-CCP,RF and IL-18 decreased significantly(P<0.01);compared with the moxibustion group,the circumference of the left and right knee joints in the NLRP3 overexpression group increased(P<0.05),the content of anti-CCP,RF and IL-18 increased(P<0.05).Conclusions Moxibustion has anti-inflammatory effects on experimental RA rabbits.Moxibustion at Shenshu(BL23)and Zusanli(ST36)can significantly reduce the level of anti-CCP,RF the immune effector,also IL-18,the in synovial fluid of RA rabbits knee joint,yet this effect can be declined significantly under NLRP3 overexpression,suggesting that moxibustion performed its anti-inflammatory regulation by suppressing abnormal immune function in experimental RA rabbits,which may be closely related to the NLRP3 expression.

Effects of Moxibustion on NLRP3 Inflammasome and Caspase-12 in Synovium of Experimental RA Rabbits / 世界科学技术-中医药现代化

Objective To observe the effects of moxibustion on the expression of Nucleotide binding oligomeric domain like receptor thermoprotein domain associated protein 3(Nod-like receptor protein 3,NLRP3)inflammasome and Cysteine aspartate protease 12(Caspase-12)in synovial tissue of experimental Rheumatoid Arthritis(RA)rabbits,and explore the anti-inflammatory mechanism of moxibustion on RA rabbits.Methods Twenty-four Japanese big-eared white rabbits were randomly divided into 3 groups according to body weight and sex:the control group,the model group and the moxibustion group.The Freund's Complete Adjuvant(FCA)was injected into the joint cavities of two knee joints of rabbits at the dose of 0.5 mL·kg-1 in the model group and the moxibustion group,and the control group was injected with the same amount of normal saline as control.For the moxibustion group,scar free moxibustion was operated at bilateral"BL 23"and"ST 36"acupoints,moxibustion for 3 times per acupoint per day,continuous treatment for 6 days,rest for 1 day,as a course of treatment,a total of 3 courses of treatment.The histological changes of synovium samples of rabbit knee joints were observed under light microscope,real-time quantitative polymerase chain reaction(RT-qPCR)was used to detect the expression of NLRP3,Cysteine aspartate protease 1(Caspase-1),Caspase-12 in synovial tissues and Enzyme-linked immuno sorbent assay(ELISA)was used to measure the change of Interleukin-1β(IL-1β)content in the synovial fluid of experimental animals.Results Compared with the control group,the degree of synovial lesions(inflammatory cell infiltration,synovial tissue hyperplasia,fibrous tissue hyperplasia,macrophage hyperplasia,etc.)and pathological score were increased(P<0.05),the expression of NLRP3,Caspase-1 mRNA were increased(P<0.05),the expression of Caspase-12 mRNA was significantly downregulated(P<0.01)and the expression content of IL-1β was significantly up-regulated(P<0.01)in the model group;Compared with the model group,the synovial lesion degree and pathological score were decreased(P<0.05),the expression of NLRP3,Caspase-1 mRNA were decreased(P<0.05),the expression of Caspase-12 mRNA was significantly increased(P<0.01)and the expression of IL-1β was decreased(P<0.05)in the moxibustion group.Conclusion Moxibustion can significantly inhibit the pathological process of knee synovitis in RA rabbits;Moxibustion at"Shenshu(BL 23)"and"Zusanli(ST 36)"acupoints can significantly up-regulate the expression of Caspase-12 and then inhibit the NLRP3 inflammasome,which may be one of mechanisms of moxibustion to reduce RA inflammatory response.

Clinical significance of high-risk human papillomavirus A9 infection in cervical cancer screening / 中国基层医药

Objective:To investigate the infection incidence of women with high-risk human papillomavirus(HPV) A9 in Quzhou, and to analyze its clinical significance in cervical cancer screening by combining with liquid-based cytology(TCT).Methods:A total of 9 010 patients were collected from January 2017 to May 2018 in the Maternal and Child Health Hospital of Quzhou.HPV detection was performed by Cervista method.In combination with TCT, cervical biopsy was carried out under vaginal microscopy.Results:Among 9 010 patients, 1 401 patients were HPV positive, and the A9 group had the highest rate for inspection(6.98%). The experiment data of the A9 group indicated that the portions of cervical intraepithelial lesions in liquid-based cytology NILM, ASC-US, LSIL, ASC-H, HSIL, SCC were 5.96%, 80.10%, 100.00%, 100.00%, 100.00%, respectively.Its coincidence rate with histology increased as liquid-based cytology increased, associated with high-grade lesions.Conclusion:A9 group infection is the most important cause of cervical intraepithelial lesions in Quzhou.High-risk HPV test, especially A9 group infection combined with TCT, can effectively improve the accuracy of cervical cancer screening.

Mosquitoes, midges and related arboviruses in southeast Sichuan province / 中华流行病学杂志

Objective To investigate the distribution patterns of mosquitoes,midges and related arboviruses in Sichuan province.Methods Blood-sucking insects were collected from houses and pens,using the ultraviolet lights.Mosquito samples were classified according to morphologic characteristics and then stored at liquid nitrogen.All samples were incubated with BHK-21 and C6/36 cells for virus isolation and then detected for their viral genes.Sequences of the virus were identified and analyzed by molecular biological software,such as BioEdit 7.0.5.3,MEGA 6.0.Results In total,17 019 mosquitoes from 3 genera and 4 species and 12 700 midges were collected from the southeast regions of Sichuan province in 2016 and 2017.Among them,79.4％ (13 519/17 019) belonged to Culex tritaeniorhynchus with 11.1％ (1 897/17 019) as Armigeres subalbatus,5.5％ (930/17 019) were Anopheles sinensis and 4.0％ (673/17 019) were Anopheles sinensis 3 virus strains that isolated from Culex tritaeniorhynchus were identified as type Ⅰ Japanese encephalitis virus.Seven pools of mosquitoes isolated from Hejiang county were identified Japanese encephalitis virus gene positive through PCR amplification.With 4 pool midges were detected positive for Akabane virus through PCR gene amplification while midges samples didn't have virus isolates.Conclusions Culex tritaeniorhynchus appeared the predominant species in the southeast regions of Sichuan.Japanese encephalitis virus transmitted by mosquitoes and Akabane virus by midges were prevalent in southeast Sichuan province.

Mosquitoes, midges and related arboviruses in southeast Sichuan province / 中华流行病学杂志

Objective: To investigate the distribution patterns of mosquitoes, midges and related arboviruses in Sichuan province. Methods: Blood-sucking insects were collected from houses and pens, using the ultraviolet lights. Mosquito samples were classified according to morphologic characteristics and then stored at liquid nitrogen. All samples were incubated with BHK-21 and C6/36 cells for virus isolation and then detected for their viral genes. Sequences of the virus were identified and analyzed by molecular biological software, such as BioEdit 7.0.5.3, MEGA 6.0. Results: In total, 17 019 mosquitoes from 3 genera and 4 species and 12 700 midges were collected from the southeast regions of Sichuan province in 2016 and 2017. Among them, 79.4% (13 519/17 019) belonged to Culex tritaeniorhynchus with 11.1% (1 897/17 019) as Armigeres subalbatus, 5.5% (930/17 019) were Anopheles sinensis and 4.0% (673/17 019) were Anopheles sinensis 3 virus strains that isolated from Culex tritaeniorhynchus were identified as typeⅠ Japanese encephalitis virus. Seven pools of mosquitoes isolated from Hejiang county were identified Japanese encephalitis virus gene positive through PCR amplification. With 4 pool midges were detected positive for Akabane virus through PCR gene amplification while midges samples didn't have virus isolates. Conclusions: Culex tritaeniorhynchus appeared the predominant species in the southeast regions of Sichuan. Japanese encephalitis virus transmitted by mosquitoes and Akabane virus by midges were prevalent in southeast Sichuan province.

Effect of Acupuncture on Spatial Learning and Memory Abilities and Hippocampal CA3 Regional BDNF in Rats with Post-stroke Depression / 上海针灸杂志

Objective To investigate the effects of acupuncture at points Baihui (GV20), Yintang (Ex-HN3) and Sishencong (Ex-HN1) on spatial learning and memory abilities and hippocampal expression of brain-derived neurotrophic factor (BDNF) in a rat model of post-stroke depression (PSD). Method Thirty male adult SD rats were allocated, using a random number table, tocontrol, acupuncture and sham operation groups, 10 rats each. A rat model of PSD was made by classical middle cerebral artery occlusion (MCAO) using an intraluminal thread technique, chronic unpredictable mild stress (CUMS) and solitary feeding. At the fourteenth day after the beginning of stress, the acupuncture group received acupuncture intervention for 14 consecutive days. At one week after the end of stress, the Morris water maze test was conducted to assess rat spatial learning and memory abilities. After the completion of the water maze test, hippocampal CA3 regional BDNF expression was determined by immunohistochemical method. Result Rat learning and memory abilities decreased significantly in the control group compared with the sham operation group (P<0.05). Space exploration time and the number of crossing platform reflecting memory abilities decreased significantly in the control group (22.36±6.20 s and 3.72±0.90 times, respectively) compared with the sham operation group (48.73±5.40 s and 8.60±1.10 times, respectively); there were statistically significant differences (P<0.01). Learning abilities, space exploration time, the number of crossing platform and BDNF expression increased significantly in PSD rats treated with acupuncture compared with the control group; there were statistically significant differences (P<0.01). Conclusion Acupuncture can improve spatial learning and memory abilities and raise hippocampal regional BDNF levels in rats with post-stroke depression.

Whole genome sequence analysis of an emerging Japanese encephalitis virus (genotype Ⅴ) strain isolated in mainland China / 中华微生物学和免疫学杂志

Abstract] Objective To analyze the molecular characteristics of the full-length genome of a new-ly isolated genotypeⅤJapanese encephalitis virus (JEV) strain (XZ0934) in China and the first genotypeⅤJEV strain ( Muar) isolated in Malaya 60 years ago.Methods Several softwares including ClustalX 2.0.9, DNAStar 7.1, Bioedit 7.2.5 and MEGA6.06 were used to conduct sequence alignments and phylo-genetic analysis.Results The full-length genomes of XZ0934 strain (isolated in Tibet, China in 2009) and Muar strain (isolated in Malaya in 1952) were composed of 10 983 and 10 988 nucleotides, respective-ly.The XZ0934 strain was highly similar with the Muar strain showing the homology of 90.6%in nucleotide (nt) and 98.3%in amino acid (aa).The open reading frame (ORF) of the two genotype Ⅴ JEV strains encoded 3433 aa while the ORF of other four genotypes (Ⅰ-Ⅳ) (10 299 nt) encoded 3432 aa.Compared with JEV strains of other genotypes, a serine were inserted into the NS4A gene of JEV strains genotype Ⅴand 10-14 nucleotides were inserted into the downstream of the ORF stop codon in 3′-untranslated region. Phylogenetic analysis of E sequences of all JEV strains genotypedⅠ-Ⅴrevealed that in the cluster of geno-typeⅤ, XZ0934 and 10-1827 ( isolated from mosquitoes in South Korea, 2010) stains formed a branch and were divergent from that of Muar strain indicating that there were molecular genetic differences among geno-typeⅤJEV strains after a 60 years hiatus.Conclusion The two genotypeⅤJEV strains showed high lev-els of identity in nucleotide sequences and amino acid sequences with serine insertion in the NS4A gene. However, there were molecular genetic differences between genotypeⅤJEV strains isolated after a 60 years hiatus.

The relationship between serum betatrophin levels of newly diagnosed type 2 diabetes patients and insulin resistance / 中国现代医生

Objective To investigate the association between serum betatrophin levels and insulin resistance in newly diagnosed type 2 diabetes patients. Methods A total of 31 healthy subjects (control group) and 35 newly-diagnosed type 2 diabetes patients(new T2DM group) were recruited. Serum betatrophin was detected with the methods of ELISA, insulin resistance was assessed with homeostasis model(HOMA-IR). Results The levels of betatrophin, free fatty acids, HOMA-IR, fasting plasma glucose, fasting serum insulin, high-density lipoprotein cholesterol and low-density lipoprotein cholesterol were significantly higher in group B than those in group A (P<0.05). Conclusion Insulin resis-tance leads to elevated serum levels of betatrophin in patients with newly diagnosed type 2 diabetes.

Assessment of N-terminal pro-brain natriuretic peptide in volume status in maintenance hemodialysis patients with non dominant edema / 中华肾脏病杂志

objective To investigate the value of NT-proBNP in assessing the volume status in maintenance hemodialysis patients with non-dominant edema.Methods One hundred and forty-five patients were recruited.Bioimpedance measurements were performed for overhydration (OH).NT-proBNP was detected by colloidal gold method.Patients were divided into three groups by levels of OH variability (△ OH,equal to OH minus weight increase) as group H (hypervolemia,n=90); group N (normovolemia,n=36) and group L (hypovolemia,n=19).Hemoglobin,albumin,blood urea nitrogen and serum creatinine were assayed,blood pressure and body mass increase were recorded.Dry weight of patients in Group H were adjusted in 3 months,the relationship between NT-proBNP and volume change were assessed.Results (1) At baseline,overall plasma NT-proBNP levels were higher than normal range.The median NT-proBNP levels in group H and group N were [1318.50(IQR 717.00,3154.25) pg/ml] and [703.50 (IQR 873.00,450.50) pg/ml],respectively.NT-proBNP was positively correlated with △OH value (r=0.801,P ＜ 0.001).(2) After 3 months,NT-proBNP levels in group H was significantly lower than baseline.Forty-one patients reached normal volume range (group H1),49 patients were resistant hypervolemia (group H2).The median NT-proBNP levels in group H1 and group H2 were [685.00 (IQR 422.50,988.50) pg/ml] and [1569.00 (IQR 982.50,2500.50) pg/ml],△ OH in group H1 and group H2 were [(0.63±0.23)L] and [(1.75±0.71)L],respectively.NT-proBNP and △ OH value in two groups had significant difference (P ＜ 0.05).NT-proBNP was positively correlated with △ OH value (r=0.684,P ＜ 0.001).(3) The area under ROC curve for NT-proBNP was 0.818,95％CI (0.733～ 0.904),P ＜ 0.001,since the absolute value of normovolemia was defined as ≤ 1.The cut off value of plasma NT-proBNP was set at 962.50 pg/ml in MHD patients with non-dominant edema,the diagnostic specificity and sensitivity were 79.6％ and 73.2％.Conclusion NT-proBNP could be used to assess volume status in MHD patients with non dominant edema.

Effects of albumin overload on HIF/HRE transcription activity in tubular epithelial cells / 中华肾脏病杂志

Objective To explore the effects of BSA on hypoxia inducible factor/hypoxia response element (HIF/HRE) transcription activity in rat tubular epithelial cells (NRK-52E) with HRE-Luc reporter plasmid.Methods Luciferin activity of NRK-52E cells incubated by a medium contained BSA in varying concentration (0,5,10,20 mg/ml) and stimulus duration (24,48,72 h) was detected by dual luciferase detecting system based on HRE-Luc reporter plasmid and HIF-1 α expression was detected by Western blotting.Results HIF/HRE transcription activity of NRK-52E cells was increased in BSA incubation group (10 mg/ml,48 h) compared with blank control (BSA 0 mg/ml,48 h) [(2.59±0.35)vs (1.03±0.09),P=0.000].HIF-1α expression of NRK-52E cells was increased in BSA incubation group (20 mg/ml,48 h) compared with blank control (BSA 0 mg/ml,24 h) [(0.052±0.010) vs (0.014±0.003),P=0.000].Conclusion Albumin can increase HIF/HRE transcription activity of TEC.

Hint1 over expression inhibits the activity of AP-1 transcription factor in HepaG2 cells / 中华普通外科杂志

Objective To study the inhibition of AP-1 transcription factor activity by Hint1 gene over expression in HepG2 cell lines. Methods The Hintl gene was amplified, and then was inserted into the pcDNA3/HA eukaryotic expression plasmid. The constructed pHA-Hint1 plasmid was confirmed by DNA sequencing. The pHA-Hint1 was transfected into the HepG2 human hepatoma cells. Semi-quantitative RT-PCR and Western-blot were used to detecte the expression of HA-Hint1. The HepG2 cells were co-transfected with pHA-Hint1 and pAP-1/Luc luciferase reporter. At 36 h after transfection, luciferase assay system was used to detect the AP-1 transcription factor activity. Results The constructed pHA-Hint1 was confirmed by DNA sequencing, pHA-Hint1 gene transduction through lipofectine induced over-expression in HA-Hint1 mRNA (t =3.89, P<0.05) and HA-Hintl protein (t=3. 12, P<0.05). Co-transfection of Hint1 gene inhibits AP-1 luciferase activity. Cotransfection with increased concentration of a pHA-Hint1 plasmid (0 μg/ml, 0. 5 μg/ml, 1.0 μg/ml, 1.5 μg/ml, 2. 0 μg/ml) produced a concentration-dependent inhibition of AP-1 transcription factor activity. At the concentration of 1.5 μg/ml, and 2.0 μg/ml, the activity inhibition reaches significant difference ( F = 72. 009, P < 0. 05 ). Conclusion Over-expression of Hintl can, at least in part, inhibit the AP-1 transcription factor activity in HepG2 cells.

Germicidal Efficacy of Garlic Solution and Its Influencing Factors on Vibrio parahaemolyticus: An Experimental Research / 中华医院感染学杂志

0.05) after storing at room temperature less than 4 hours.Its germicidal rate decreased obviously after storing at room temperature more than 8 hours.The(germicidal) efficacy was enhanced with the up of temperature and down of pH,and sharply down with the existence of organic substances.(CONCLUSIONS) Garlic solution can kill the V.parahaemolyticus.Its storing time as well as the temperature,pH,and organic substances can influence the germicidal efficacy.

Oligo-chips for Detecting Pathogens in Cerebrospinal Fluids:A Preliminary Study / 中华医院感染学杂志

OBJECTIVE To develop a macroarray method to detect pathogens in cerebrospinal fluid.METHODS According to the bacterial 16S rRNA genes,designed 10 kinds of specific probes and a pair of universal primers that can amplify rRNA gene of all bacteria.The tailed probes were spotted onto a nylon membrane.DNA was isolated from each pathogen,and subjected to UP-PCR to amplify target fragments,which were labeled with bio-16-dUTP at the same time.All those denatured fragments were hybridized to the probes on nylon membrane and visualized by AKP labeled avidin.The sensitivity and specificity of the system were detected.A total of 32 CSF samples,which were verified the bacterial infection by the routine method,were tested by this method.RESULTS It was sensitive to 10 CFU/ml when detecting Escherichia coli.Every kind of pathogens only reacted to its corresponding probes fixed on nylon membranes,which showed high specificity.The result of identifying 32 CSF clinical specimens accorded with that of routine method.CONCLUSIONS The method can screen out common pathogens in CSF sensitively and exactly.

Inhibitory Effects of Water First Extractions of Tea on Digestive Bacteria / 中华医院感染学杂志

OBJECTIVE To study the inhibitory effects of the water extractions from ordinary green tea,black tea,chrysanthemum tea,oolong tea and Puer tea on digestive tract bacteria.METHODS Incubating plate with foveolae was used to study the inhibitory effect on Vibrio parahaemolyticus,Escherichia coli,Salmonella and Shigella with the water extractions of the tea.RESULTS It is showed inhibitory effect on V.parahaemolyticus by the first water extraction from ordinary green tea,black tea,chvysanthemum tea,oolong tea and Puer tea.Same effect was also observed to E.coli,Salmonella and Shigella.The green tea and Puer tea showed best inhibitory effects on V.parahaemolyticus,and the inhibitory zone diameter of the first water extraction of 2.5mg/ml teas were 10.6mm and 12.3mm,respectively.CONCLUSIONS The first water extraction of many kinds of the tea showes inhibitory effects on the digestive bacteria.

Statistical analysis of articles, citations and authors in Chinese Journal of Neurology from 2002 to 2004 / 中华神经科杂志

Objective To study the characteristics of articles, citations and authors in Chinese Journal of Neurology(Chin J Neurol). Methods The date about articles, citations and authors of Chin J Neurol from 2002 to 2004 were collected and analyzed in terms of bibliomertic parameters. Results There were 648 papers published in Chin J Neurol during 3 years and papers with funded projects accounted for 25.0%. The average number of articles per isseu was 36; the average number of article per page was 0.43; the average publishing cycle of the articles was 10 months. The authors distributed in 27 mainland provinces and Hongkong, 91.5% of whom shared with co-author; the coporation rate was 4.19. The papers with citation accounted for 65.3% and 95.0% of all the citations came from journals. The Price’s indicator was 56.6%. Conclusions Chinese Journal of Neurology, an academic journal of high quality, with strong influence, pushes the subject of neurology in China forward.

Relationships between NO, NOS,TNF-? in serum and surgical infection / 中国现代普通外科进展

Objective: To study the relationship between NO, NOS and TNF-? of serum and surgical infection. Methods: We used chemistry technique and ELISA to study serum NO, NOS and TNF-? in 30 cases of surgical infection.Results: Compared with normal group and before treatment group NO, NOS and TNF-? were singnificantly higher in surgical infection(P