Methodological study on eliminating nucleic acid contamination in molecular diagnostic laboratory / 生物工程学报

Nucleic acid detection technique has good sensitivity and specificity and is widely used in in vitro diagnosis, animal and plant commodity quarantine, forensic identification, and other fields. However, it is susceptible to carryover contamination during the operation and leads to false-positive results, which seriously affects the detection accuracy. Therefore, finding an effective solution to prevent and eliminate nucleic acid carryover contamination has become particularly urgent. This study compared several different methods for removing nucleic acid contamination and confirmed that sodium hypochlorite solution and PCRguard reagent could effectively eliminate nucleic acid carryover in the liquid and on surfaces of different materials. Besides, the combination of sodium hypochlorite solution and PCRguard can solve the nucleic acid aerosol contamination. This study proposes solutions for the routine prevention of carryover contamination and removal of aerosol that has occurred in molecular diagnostic laboratories.

Formulation of Criterion for Clinical Application of Shuxuetong Injection by Clinical Pharmacists in Our Hospital Based on Evidence Quality Evaluation Method / 中国药房

OBJECTIVE： To provide reference for strengthening clinical application of key monitoring drugs and promoting rational drug use in clinic. METHODS： Based on evidence-based medicine， taking key monitoring drugs Shuxuetong injection as example， clinical evidence of domestic and foreign clinical studies were collected. The included literatures were graded according to the quality of GRADE evidence and recommended strength system. Evidence-based medicine evidence for the indications of Shuxuetong injection were evaluated， and criterion for clinical use of Shuxuetong injection was formulated in Huaihua First People’s Hospital （our hospital）. RESULTS： The main content of criterion for clinical application of Shuxuetong injection formulated by our hospital was that there was A-level evidence support for acute ischemic cerebral infarction， but it was weakly recommended and only used for adjuvant therapy； there was B-level evidence support for anticoagulation （for preventing DVT）， diabetic peripheral nerve lesion， but it was weakly recommended； there was only C-level or D-level evidence support for other indications， it was strongly recommendation against use. CONCLUSIONS： Clinical pharmacists formulate the criterion for clinical application of Shuxuetong injection by evidence quality evaluation method， provide reference for clinical application management of key monitoring drug and play an important effect on rational drug use in clinic.

Spatial distribution and influencing factors among middle-aged and elderly patients with hypertension / 预防医学

Objective @#To investigate the spatial distribution and the influencing factors of hypertensive patients aged 45 years and above,and to provide evidence for the prevention and control of hypertension among middle-aged and elderly population.@*Methods @#Multi-stage stratified random sampling method was used to collect 150 counties（cities or districts）from 30 provinces（municipalities or autonomous regions）from CHARLS database in 2015 according to the per capita GDP in different areas. Then the probability proportionate to size sampling method was used to select three villages（neighborhood committees）,80 families were randomly selected from each village（neighborhood committees）,and residents aged 45 years and above were interviewed. Spatial analysis was based on the vector map of China's provincial boundaries,Geoda and ArcGIS 10.2 software were used to analyze the prevalence and influencing factors of hypertension at provincial levels.@*Results @#The health data of 14 880 people aged 45 years and above in 28 provinces（municipalities or autonomous regions）were obtained,including 4 203 patients with hypertension（28.25%）and 5 408 people with pre-hypertension（36.34%）. The results of local spatial analysis showed that the clustering regions of hypertension patients were in northeast and eastern parts of China. The results of spatial regression analysis showed that males,age of 80 years and above and overweight were the main risk factors for hypertension（all P<0.05）,which had greater influence on the middle part of China,north and northeast of China,and the east and north of China,respectively.@*Conclusion @#Males,age of 80 years and above and overweight were the main risk factors for hypertension in middle-aged and elderly population in China. The main prevention and control areas of hypertension were eastern and northeast China.

The levels of serum interferon gamma-induced protein 10 in patients with acute pathogen infection / 国际检验医学杂志

Objective To observe the levels of interferon gamma-induced protein 10(IP-10)in children with acute viral and bacterial infection.Methods There were three groups:acute viral infection group(51 ca-ses),acute bacterial infection group(52 cases),and healthy control group(51 cases).Serum IP-10 levels were detected by enzyme linked immunosorbent assay(ELISA),and serum C reactive protein(CRP)were deter-mined with BNⅡ automatic protein analyzer.Results The level of serum IP-10 and CRP were significantly different in different groups(P<0.05).Compared with control group,the level of IP-10 and CRP were higher in acute viral infection group or in acute bacterial infection group(P<0.05).The level of IP-10 was higher in acute viral infection group than that in acute bacterial infection group(P>0.05).The level of CRP was higher in bacterial infection group than that in acute viral infection group(P<0.05).The IP-10 levels in viral Infec-tion patients were not correlated with the CRP levels.The area under curve(AUC),sensitivity and specificity of IP-10 and CRP were 0.688 and 0.873,35.0% and 79.6%,96.1% and 98.0%.When cut-off value of predic-tive probability was 0.713,sensitivity and specificity were increased to 82.5% and 100.0%.Conclusion Ser-um IP-10,CRP and predictive probability are valuable in the diagnosis of acute pathogen infection in children.

Value of serum IgG4 level for differential diagnosis of IgG4-related hepatobiliary diseases and other hepatobiliary diseases / 临床检验杂志

Objective To investigate the value of serum IgG4 level for the diagnosis of IgG4-related hepatobiliary diseases and the differentiation from other hepatobiliary diseases.Methods A total of 270 patients with hepatobiliary diseases in the People's Hospital of Hunan Province from August 2015 to April 2017 were enrolled in this study,and 20 healthy subjects were selected as controls.The 270patients were divided into eight groups:liver cirrhosis group (n =17),acute pancreatitis group (n =52),chronic pancreatitis group (n =33),cholecystitis and gallstone group (n =27),bile duct carcinoma group (n =30),cholangitis and biliary calculi group (n =41),pancreatic cancer group (n =47),IgG4-related hepatobiliary disease group (n =23).The levels of serum IgG4 were measured by rate nephelometery assay.The sensitivity and specificity of IgG4 levels for distinguishing IgG4-associated hepatobiliary diseases were evaluated by receiver operating characteristic curve.Results The levels of IgG4 of the cirrhosis group and the IgG4 related hepatobiliary disease group were significantly higher than those of the control group (P ＜ 0.05).The IgG4 level in the hepatobiliary disease group was significantly higher than those of the other seven groups (Z =-5.267,-6.802,-5.921,-6.005,-6.173,-6.513,-6.014,P all ＜ 0.01).The area under curve (AUC) for IgG4 level in distinguishing IgG4 associated hepatobiliary diseases and other hepatobiliary diseases was 0.982.When 4.13 g/L was used as the cut off value of diagnosis,the sensitivity and specificity of IgG4for diagnosis were 95.7％ and 96.0％ respectively.The IgG4 levels in twelve patients with IgG-associated hepatobiliary diseases after 2 months of glucocorticoid therapy were significantly lower than those before glucocorticoid therapy (Z =-2.021,P =0.043).Conclusion The elevated serum IgG4 level may not be specific just for IgG4-related hepatobiliary diseases.The cut off value of 4.13 g/L should be very useful for diagnosing IgG4-related hepatobiliary diseases,differentiating from other hepatobiliary diseases and evaluating the therapeutic effect of glucocorticoid therapy.The further detailed verification for these findings should be necessary in clinical practice by increasing the sample size.

The clinical research about early diagnosis of small cell lung cancer through mad2 de-tection / 中国肿瘤临床

Objective:To explore combined detection of mad2 with anti-nuclear mitotic spindle apparatus antibody(MSA)and anti-centromere antibody(ACA)and their clinical value for the diagnosis of small cell lung cancer(SCLC).Methods:One hundred and twen-ty SCLC patients,110 non-small cell lung cancer(NSCLC)patients,and 115 pulmonary nodule(PN)patients were enrolled in this study. The expression of mad2 was analyzed by qt-PCR.MSA and ACA were detected by indirect immunofluorescence(IIF)staining.Results:mad2 was overexpressed in SCLC and NSCLC samples(P<0.05).There were significant differences between the results obtained for SCLC and NSCLC samples by qt-PCR(P<0.05).AUC in ROC curve for mad2 expression was 0.799 with an intermediate diagnostic value. In the correlative analysis,the odds ratio of MSA and ACA was 6.94 and 5.60,respectively.In the correlation analysis,Kappa value of mad2 with MSA was 0.49,and Kappa value of mad2 with ACA was 0.42.In the parallel analysis,the sensitivity and specificity was 83.31% and 79.34%,respectively,while the Youden Index was 0.62.Moreover,in the serial analysis,the sensitivity and specificity was 65.32% and 93.35%,respectively,and the Youden Index was 0.59.Conclusions:In comparison with the NSCLC and PN samples,mad2 was overexpressed in SCLC samples.Therefore,mad2 ought to play a critical role in the pathology of SCLC.The combined expression of mad2 with MSA and ACA may contribute to enhancing the sensitivity and specificity of detection;this expression may allow early diag-nosis and clinical diagnosis of SLCC and may be a promising treatment for SCLC.

Comparative observation of the clinical effect of immediate recovery of fluid resuscitation in traumatic shock and delayed resuscitation / 中国综合临床

Objective To investigate the clinical value of immediate fluid resuscitation and delayed resuscitation in patients with traumatic shock. Methods The patients with traumatic shock treated in the Critical Care Medicine Department of People's Hospital of Wenjiang District from March 2014 to March 2017 were selected. According to the number of admission cases,one hundred and twenty patients with traumatic shock were randomly divided into two groups,60 cases in each group. The control group was given early immediate fluid resuscitation,the observation group was given delayed resuscitation,and the blood coagulation and blood routine indexes of the two groups were compared before and after the fluid resuscitation in the two groups,and the amount of fluid rehydration and the fatality rate in the two groups of patients with 1 h shock were observed,and the incidence rate of acute respiratory distress syndrome ( ARDS) and multiple organ dysfunction syndrome (MODS) were compared. Results After treatment,the blood clotting and blood routine indexes of the two groups were improved (P<0. 05),of which the thromboplastin time (PT) ((11. 04±1. 17) s),activated partial thromboplastin time (APTT) ((28. 12±5. 93) s) in the observation group in the observation group were lower than those of the control group( (15. 12±1. 26) s,(36. 17±9. 05) s) (t = -15. 37,-9. 81,P<0. 05),platelet countPLT) ((146. 92±16. 85)×109 / L) was higher than that of the control group ((114. 18±10. 69)×109 / L ) (t= -9. 77,P<0. 05),and the blood routine hemoglobin (Hb) ((112. 21±9. 46) g/ L),and the base surplus (BE)((-5. 30 ± 2. 45) mmol/ L ) were all higher than those of the control group ((92. 95 ± 11. 20) g/ L, (-8. 27±3. 53) mmol/ L ) (t= -11. 46,-8. 99,P<0. 05),blood lactic acid (BL) ((2. 79±1. 12) mmol/ L ) was lower than that of the control group ((3. 54±1. 37) mmol/ L) (t = -8. 99,P<0. 05). The volume of 1 h infusion of shock in the observation group ((569. 96±187. 34) ml ) was lower than that of the control group((1957. 35±204. 14) ml) (t = 8. 725,P<0. 05). The incidence of ARDS (3. 33% (2/ 60)),MOD(3. 33%(2/ 60)) and fatality(1. 67%(1/ 60)) were lower than those of the control group(8. 33%(5/ 60),6. 67%(4/60),6. 67%(4/ 60) ( χ2 = 2. 725,3. 214,2. 985,P< 0. 05) . Conclusion The early stage of traumatic shock delayed fluid resuscitation is conducive to the protection of the blood coagulation function of patients,to improve blood indicators,to reduce the amount of 1 h infusion and to reduce the incidence of ARDS and MODS.

Clinical significance of PCT,DD,and CRP levels in patients with infection in acute-on-chronic liver fail-ure / 实用医学杂志

Objective To investigate the clinical diagnostic value of serum procalcitonin(PCT),D-dimer (DD),C-reactive protein(CRP)in acute-on-chronic liver failure(ACLF). Methods 124 ACLF patients, 63 chronic hepatitis B patients,32 chronic hepatitis C patients,24 chronic hepatitis E patients and 60 healthy controls from the second affiliated hospital of Nanchang University were enrolled in this study.PCT was detected by a sandwish immunodetection method. D-dimer was detected by Latex Turbidimetry. CRP was detected by rate nephenometry. The detection results were used for analyzing the clinical diagnostic value of ACLF with infection. Results(1)The level of PCT,DD and CRP in ACLF group were significantly higher than non-ACLF group and healthy controls(P<0.05).The levels of PCT,DD and CRP in the infection group were significantly higher than non-infection group(P<0.05).(2)The positive rates of PCT,DD and CRP in the infection group were 93.24%, 78.38%,89.19%,which were significantly higher than the non-infection group and healthy controls respectively (P < 0.05).(3)The sensitivity(93.24%)and specificity(90.00%)of PCT were the highest among all indexes. (4)The area under the ROC curve of PCT,DD,CRP were 0.892,0.715,0.755,respectively.PCT had the highest diagnostic value. Conclusion The levels of serum PCT,DD and CRP have a significant clinical value for the early diagnosis of ACLF with infection.

Influence of atorvastatin to TNF-α and IL-10 in intracerebral hemorrhage rats / 中国基层医药

Objective To observe the neuroprotective effect of atorvastatin and its influence to tumor necrosis factor-alpha(TNF-α) and intedeukin-10(IL-10)in intraeerebral hemorrhage(ICH)rats,and to explore the possible mechanism of its neuroproteetive effect.Methods ICH WaS induced by stereotaxic infusion of collagenase into the right baSal ganglia,the expressions of TNF-α,IL-10 in the serum and brain tissue were analyzed by enzyme linked immunosorbent assay(ELISA),HE staining was used to evaluate the number of activated microglia.Results (1)The numbers of activated mieroglia,TNF-α and IL-10 were have significant differences between group A and group B(all P＜0.05).(2)In comparision with group B,numbers of activated mieroglia in group C,D and E were significantly redueed(P＜0.05).Atorvastatin cause a dose-dependent regulation,which decreased the level of TNF-α and inereased the level of IL-10 significantly(all P＜0.05).Conclusion The down-regulation of TNF-α and up-regulation of IL-10 might partly attribute to the neuropmtection of atovastatin.

Experimental parameters for evaluating the functional prognosis of systemic scleroderma / 中国组织工程研究

BACKGROUND: Antinuclear antibody, anticentromere antibody, anti-cytoplasm antibody, and antibody against SCL-70 are the main self-antibodies involved in systemic scleroderma (SSc) and are closely connected with the development of SSc.OBJECTIVE: To probe the value of various self-antibodies and proteins in evaluating the functional prognosis of patients with SSc. DESIGN: Case-control study.SETTING: Clinical Laboratory of Second Hospital Affiliated to Jiangxi Medical College.PARTICIPANTS: Totally 74 patients, 19 males and 55 females aged 12-59years old, were confirmed of SSc at the outpatient and inpatient departments of Second Hospital Affiliated to Jiangxi Medical College between December 1995 and December 2004. There were 46 cases of diffuse cutaneous SSc, 24 cases of localized cutaneous SSc, and 4 cases of overlapping syndrome. Meanwhile40 inpatients (14 males and 26 females aged 19-54years old) who received treatment due to other diseases were recruited from the same hospital.METHODS: The level of antinuclear antibody, anticentromere antibody,and anti-cytoplasm antibody was detected with indirect irnmuneofluorescence assay; antibody against SCL-70 was detected with Western blot; the level of serum immunoglobulin, C-reactive protein (CRP) and rheumatoid factor was examined with velocity dispersion turbidimetry.ticentromere antibody, anti-cytoplasm antibody, and antibody against SCL-70;RESULTS: Blood samples collected from the 74 patients with SSc and 40controls were proved eligible and all data entered the final statistical analanti-cytoplasm antibody, and antibody against SCL-70: It was obviously higher in SSc group than in control group [66% (49/74), 53% (39/74), 39%(29/74), 7% (5/74), 0, (x2=57.15, P ＜ 0.01)]. The positive rate of antinuclear antibody in patients with diffuse cutaneous SSc was significantly lower than in patients with localized cutaneous SSc [57% (26/46), 83%(20/24),(x2=5.03, P ＜ 0.05)], but the positive rate of antibody against SCL70 was significantly higher than localized cutaneous SSc [48% (22/46),rheumatoid factor in patients with SSc was markedly higher thanin control group [(16.89±11.94), (11.89±2.05) g/L; (23.06±6.18), (22.44±5.53) IU/mL,t=8.01, 2.46, P ＜ 0.01].CONCLUSION: The positive rate of antinuclear antibody, anticentromere antibody, anti-cytoplasm antibody, and antibody against SCL-70, as well as the level of immunoglobulin G and rheumatoid factor were obviously increased in patients with SSc, suggesting that these experimental parameters have the value in evaluating the prognosis of SSc.

The effect and mechanism of caveolin-1 upregulation on the proliferation of MGC803 cell line / 中国医师杂志

Objective To study the effect of caveolin-1 gene expression on the proliferation of human gastric adenocarcinoma cells,and to explore the possibility for its future usage in gene therapy.Methods The full-length caveolin-1 gene was stably transfected into the MGC803 cell line by lipofectin.The Pcl neo vector was transfected at the same time as mock control.The expression of caveolin-1 was detected by Western blot in both the caveolin-1 gene transfected MGC803 cells and the controls.The cell cycle was analyzed by flow cytometry.Results After transfected with caveolin-1,MGC803 cells significantly up-regulated the expression of caveolin 1 and extended their doubling time.The cell proliferation was inhibited and the cell cycle was arrested in the G_0/G_1 phase.Conclusion Caveolin-1 can inhibit the proliferation of MGC803 cells and induce cell cycle arrest in G_0/G_1 phase.

Preventive effects of curcumin on status epilepticus induced by lithium chloride-pilocarpine / 中国医师杂志

Objective To investigate the preventive effects of curcumin for status epilepticus(SE) induced by lithium chloride-pilocarpine.Methods Totally 45 Sprague-Dawley(SD) rats were randomly divided into three groups: preventive group(n=15),non-preventive group(n=15),and control group(n=15).The latency peroid and incidence of SE were recorded.The surviving neurons were stained by using nissl staining,and the programme death cells were detected by using terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL) in hippocampal CA3.Results The SE incidence of preventive group was 66.7%,which was significantly lower than that of non-preventive group(P0.05).Conclusion Pretreatment of curcumin can prevent the SE induced by lithium chloride-pilocarpine and the pretreatment can not protect the neuron.

Knockdown of Survivin Gene Expression in Neuroblastoma Cells by RNAi / 中国医师杂志

Objective To construct the short hairpin RNA(shRNA) expression vector of survivin and down-regulate the expression of survivin through RNA interference in neuroblastoma cell line SH-SY5Y. Methods Two pairs of oligonucleotide sequences specific for human survivin mRNA were designed and synthesized. The annealed oligonucleotide fragments were subcloned into pBSHH1 plasmid. After being identified by restriction enzyme digestion and sequencing, the recombinant plasmids pBSHH1-S1 and pBSHH1-S2 were transfected into SH-SY5Y cells, respectively. Survivin expression in the transfected cells was assayed by both RT-PCR and western blot. Results Enzyme digestion analysis and DNA sequencing showed that the oligonucleotide fragments were correctly inserted into pBSHH1 plasmid, and survivin expression in the transfected cells was knocked down significantly by pBSHH1-S1 or pBSHH1-S2 at both the protein and mRNA level. Conclusion The shRNA expression vectors of survivin were successfully constructed, and could down-regulate survivin expression in SH-SY5Y cells, which lay a foundation for further research on gene therapy for tumors such as neuroblastoma.

Expression of ubiquitin in neurofibrillary tangles and senile plaque neurites in hippocampal formation of Alzheimer's disease brain / 中华神经科杂志

Objectives To detect the expression and distribution of ubiquitin in neurofibrillary tangles(NFT) and senile plaque neurites in the hippocampal formation of Alzheimer's disease brain and explore the significance of ubiquitin in pathological mechanism of Alzheimer's disease.Methods The expression of ubiquitin in the hippocampal formation of 7 patients with Alzheimer's disease and 5 non-dementia elderly was detected using immunohistochemical technique.Results Neurofibrillary tangles and senile plaque neuritis were labeled by the antibody of ubiquitin in hippocampal formation of Alzheimer's disease brain, but pretangle neurons were not stained. Containing NFT neurons stained by ubiqutin antibody immunoreactive products were distributed more in the CA1 and CA2 (42.13?0.65, 30.57?0.78 respectively)than those in CA3 and CA4 (12.43?0.24?18.34?0.81 respectively)of the hippocampal formation (P

Value of differential diagnosis of 5-hydroxytryptamine,dopamine and their metabolic productions in cerebral spinal fluid in patients with viral encephalitis and schizophrenia / 临床神经病学杂志

Objective To investigate the value of differential diagnosis of 5-hydroxytryptamine,dopamine and their metabolic productions in cerebral spinal fluid(CSF) in patients with viral encephalitis and schizophrenia.Methods The CSF concentrations of DA,5-HT,5-hydroxyindole acetic acid(5-HIAA) and homovanillic acid(HVA) were assayed by high-performance liquid chromatography(HPLC) in the patients with viral encephalitis and meningoencephalitis(67 cases),schizophrenia(37 cases) and other diseases(21 cases).Results(1) Compared with case control group,the concentrations of 5-HT,5-HIAA and the ratios of 5-HIAA/5-HT,5-HT/DA were significantly lower in viral encephalitis and meningoencephalitis group(P

Effects of long-term replacement therapy of estrogen or compound estrogen on the deposition of beta-amyloid protein in OVX rat hippocampal formation / 临床神经病学杂志

Objective To investigate the effects of long-term replacement therapy of estrogen or compound estrogen on the deposition of beta-amyloid protein (A?) in ovariectomy (OVX) rat hippocampal formation. Methods Fifty seven-month-old female Sprague-Dawley rats were randomly divided into five groups: normal control, SHAM, OVX, OVX plus 17beta-estradiol (OVX/ERT), and OVX plus compound nylestriol tablet (OVX/NL). The ovariectomy was performed in OVX, OVX/NL and OVX/ERT rats and drugs were administered orally in OVX/NL and OVX/ERT rats for 35 weeks after the ovariectomy. Then the rats in all groups were sacrificed. Immunohisto-chemistry of A?, cell counting and imaging system were used to determine the change of A? deposition levels in OVX rat hippocampal formation. Results The numbers and optical density of A?-positive neurons of all hippocampal subregions and dentate gyrus in OVX rats were markedly higher than those of normal control, SHAM,OVX/NL, and OVX/ERT rats( P