Establishment and application of miRNA detection method for forensic body fluid identification / 中国法医学杂志

Objective To establish a SYBR Green real-time PCR detection method with tissue-specific miRNAs and explore a novel approach for forensic body fluid identification. Methods The frequently reported 6 standard miRNAs were synthesized to establish a SYBR Green method, and verify with body fluid. The relative expression data for the 6 miRNAs were obtained using SYBR Green real-time PCR method in peripheral blood, menstrual blood, saliva and semen. Results The assays showed that miRNA205 permitted the unequivocal identification among different fluids. miRNA451 and miRNA144 could be used to distinguish blood from non-blood. Menstrual blood or peripheral blood could be identified through miRNA214. miRNA888 and miRNA891 was highly expressed in semen. Conclusion The results of this study indicate that miRNA SYBR Green profiling may provide a feasible and effective approach to body fluid identification for forensic casework.

A study on screening,treatment and prognosis of congenital hypothyroidism in Xuancheng City / 中国新生儿科杂志

Objective To review the experience on screening,diagnosis and treatment for neonatal congenital hypothyroidism (CH)in Xuancheng City,Anhui Province.MethodsFrom Jan 2008 to Dec 2014,peripheral blood samples from the heel were collected among 83787 neonates and the blood samples were sent to the screening center to test the level of thyroid stimulating hormone (TSH).Neonates with abnormal level of TSH were admitted to our hospital for further TSH and free tetraiodothyronine (FT4 )tests. Neonates with CH diagnosis were given standard treatments of levothyroxine and their thyroid function, physical and intelligence development were evaluated on a routine basis.Results Among the 106 neonates with abnormal TSH level ,68 were diagnosed CH.The prevalence of CH in Xuancheng City was 0.8 ‰.All 68 patients with CH received levothyroxine treatment and 6 cases were lost during follow-up.44 patients received physical examination and their bone ages were normal. 53 patients received intelligence assessment.Only 1 of them was diagnosed of mental retardation, and 3 with suspicious mental retardation.Conclusion Neonatal screening is necessary for the early diagnosis and treatment of CH. Neonates with CH can have normal physical and mental development if they receive standard treatment as early as possible and followed up regularly.

The clinical effect of shuxuening treatment on organophosphorus pesticide poisoning with toxic myocarditis / 中国基层医药

Objective To investigate the clinical effect of shuxuening treatment on organophosphorus pesti-cide poisoning with toxic myocarditis.Methods 60 patients with organophosphorus toxic myocarditis were selected in our hospital as the research subjects,and 60 cases were divided into two groups:observation group(n =30) and control group(n =30).Control group was given conventional treatment,treatment group was given shuxuening injec-tion 14 days on the basis of conventional treatment.After treatment,creatine kinase isoenzyme (CKMB),troponin (TNI),interleukin 6 (IL -6) and cholinesterase(ChE) were compared,and the changes of clinical symptoms were observed at the same time.Results There was no significant difference between the observation group and the control group(χ2 =0.630,P =0.730).The TNI,IL -6,CKMB could reflect the severity of myocardial injury in patients with different degrees of organic phosphorus poisoning,TNI,CK -MB,IL -6 increased with the degree of poisoning,the differences were statistically significant(F =11.863,4.512,3.774;P =0.000,0.015,0.029).After treatment for 4, 9,14 days,TnI,CK,CK -MB,levels of IL -6 in the two groups were recovered,but the recovery levels of TnI,CK -MB and IL -6 of the observation group significantly better than the control group,the differences were statistically sig-nificant(Fourth days,t =8.125,5.128,10.461;P =0.000,0.001,0.000;Ninth days,t =5.464,4.674,9.510;P =0.001,0.002,0.000,t =6.162,8.248,5.523;P =0.000,0.000,0.001).Conclusion Conventional treatment combined with shuxuening in the treatment of organophosphorus pesticide poisoning with toxic myocarditis has better therapeutic effect,it is worthy of promotion.

The correlation analysis of indoor carbon monoxide concentration and chronic diseases / 国际检验医学杂志

Objective To explore the correlation of the indoor carbon monoxide concentration and chronic diseases by monitoring the indoor carbon monoxide concentration timely.Methods Collected the data of non-traumatic patients(n=80)from July 201 1 to January 2014,and assigned them into the experiment group with carbon monoxide concentration measured and the control group without measured.All the subjects were further divided into one group with complaint of carbon monoxide poisoning and the other one without.For patients exposed to high concentration of carbon monoxide,carboxyhemoglobin was also measured and APACHEⅡ was scaled,and the relationship between them was analyzed.The confirmation and misdiagnosis rate of carbon monoxide poison-ing were calculated.For the patients with acute exacerbation of chronic diseases like chronic cardiac failure,chronic obstructive pul-monary disease,cerebral infarction induced by carbon monoxide poisoning,BNP,CAT and NIHSS were documented on admission and during follow-up with removal of carbon monoxide exposure and compared respectively.Results The relationship between blood carboxyhemoglobin,APACHEⅡ scores on admission and indoor carbon monoxide concentration was linear,and obviously positive.Between the experiment group(n=40)and the control group(n=40),there was significant difference(P <0.05)in confirma-tion rate and misdiagnosis rate with 80.00% vs.55.00% and 6.25% vs.81.80% respectively.For the 54 patients with carbon monoxide poisoning diagnosed,the changes between before and after removal of carbon monoxide exposure of the CAT scores of chronic obstructive pulmonary disease and NIHSS score of CI disease and myoglobin,troponin I,creatine kinase isoenzyme,BNP for chronic cardiac failure were significantly different(P <0.01).Conclusion The indoor carbon monoxide concentration may indicate the severity of carbon monoxide poi-soning,which could increase the confirmation rate of carbon monoxide poisoning,and reduce the misdiagnosis rate.It is helpful to identify carbon monoxide exposure,the common inducing factor,so as to improve the patients′clinical symptoms.

Compound K suppresses myeloid-derived suppressor cells in a mouse model bearing CT26 colorectal cancer xenograft / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of ginseng-derived compound K (C-K) on apoptosis, immunosuppressive activity, and pro-inflammatory cytokine production of myeloid-derived suppressor cells (MDSCs) from mice bearing colorectal cancer xenograft.</p><p><b>METHODS</b>Flow-sorted bone marrow MDSCs from Balb/c mice bearing CT26 tumor xenograft were treated with either C-K or PBS for 96 h and examined for apoptosis with Annexin V/7-AAD, Cox-2 and Arg-1 expressions using qRT-PCR, and supernatant IL-1β, IL-6, and IL-17 levels with ELISA. C-K- or PBS-treated MDSCs were subcutaneously implanted along with CT26 tumor cells in WT Balb/c mice, and the tumor size and morphology were evaluated 21 days later.</p><p><b>RESULTS</b>C-K treatment significantly increased the percentages of early and late apoptotic MDSCs in vitro (P<0.01 and P<0.05, respectively), decreased the expressions of immunosuppression-related genes Cox-2 (P<0.05) and Arg-1 (P<0.01), and suppressed the production of IL-1β (P<0.05), IL-6 (P<0.01), and IL-17 (P<0.05) by the MDSCs . Compared with PBS-pre-treated cells, C-K-pretreated MDSCs showed significantly attenuated activity in promoting CT26 tumor growth in mice (P<0.01).</p><p><b>CONCLUSION</b>C-K can suppress the immunosuppresive effect of MDSCs to inhibit tumor cell proliferation in mice, which suggests a new strategy of tumor therapy by targeting MDSCs.</p>

Clinical efficiency of deferred limited TURP in the treatment of urinary retention after 125I seed implantation / 中华泌尿外科杂志

Objective To evaluate the clinical efficiency and complications of deferred limited TURP for treating urinary retention after 125I seed implantation.Methods From Jan.2006 to Jan.2014,36 prostate cancer patients with severely dysuria or retention were performed with deferred limited TURP 6 months after 125I seed implatation.The average age was 66 (57 to 82) years.The average PSA was 8.5 (3.5 to 25.6) μg/L before seed implantation.The average prostate volume was 78 (45 to 110) ml.Before limited TURP,the average IPSS was 16.5 (13 to 32),the average QOL score was 5.5 (5 to 6),the Qave was 5.6 (2 to 9) ml/s,the average PVR was 285 (120 to 550) ml.The urination state,QOL and complications were evaluated the second day after catheter removal and one,three and six months after limited TURP.Results Limited TURP was successfully performed in all 36 patients.The average operation time was 45 (35 to 60) min.The average fellow-up time was 42 (6 to 84) mon.The second day after catheter removal,the average IPSS was 4.5 (3 to 6),the average QOL score was 2 (1 to 3),the Qave was 14.5 (12 to 21) ml/s,the average PVR was 35 (20 to 50) ml.One month later,the average IPSS was 3.5 (2 to 5),the average QOL score was 2.0 (1 to 3),the Qave was 15.5 (12 to 23) ml/s,the average PVR was 30 (20 to 40) ml.Three months later,the parameters continued to improve and stabilized.The second day after catheter removal and one,three and six months after limited TURP,all parameters had significant improvement compared with those before limited TURP with statistical significance (P ＜ 0.05).Four cases had mild incontinence,no case had urethral ischemia and necrosis.Conclusions 6 months after 125I seed implatation,prostate cancer patients with severely dysuria or retention can be safely and effectively treated with limited TURP.

Establishment of an anti-CD45 monoclonal antibody that inhibits T-cell proliferation / 中国实验动物学报

Objective To identify molecules that modulate T-cell functions and serve the studies on T-cell media-ted autoimmune diseases.Methods Bone marrow-derived dentritic cells were collected from BALB/c mice to immunize Wistar rats, and to establish many hybridoma cell lines.Many hybridoma cell lines which could modulate T-cell functions were obtained.One of the cell lines, most actively inhibiting T-cell proliferation, was further studied.Results The anti-CD45 mAb recognized CD45 and significantly suppressed T-cell proliferation in proliferation assays.Conclusions Our re-sults indicate that the anti-CD45 mAb can effectively suppress T-cell proliferation, and is promising to be used in the pre-vention and treatment of T-cell mediated autoimmune diseases in the future.

Evolution of hepatitis C virus quasispecies during natural disease progression of chronic hepatitis C and the clinical implications / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the variations of hepatitis C virus (HCV) quasispecies and the changes in their composition in untreated patients with chronic hepatitis C.</p><p><b>METHODS</b>Eleven patients chronic hepatitis C without previous specific anti-HCV treatment were tracked for disease progression and blood samples were collected at multiple time points. The major clinical parameters of liver function and viral load were tested. A fragment of HCV hypervariable region 1 (HVR1) was amplified and cloned, and the positive clones were sequenced and subsequently analyzed to determine the composition variation of HCV quasispecies during disease progression in relation to the major clinical parameters.</p><p><b>RESULTS</b>A total of 631 HVR1 sequences were acquired from the positive clones. The evolution of HCV HVR1 quasispecies in untreated chronic hepatitis C patients featured 3 patterns of variation in quasispecies composition, namely stable, fast and slow changes during the natural course of chronic hepatitis C. The genetic distance of the quasispecies was found to inversely correlated with ALT (R=-0.438, P=0.011) and AST level (R=-0.500, P=0.003), and sense mutation rate was also inversely correlated with ALT level (R=-0.387, P=0.026) and AST level (R=-0.410, P=0.018). No significant association was found between HCV load and any clinical or virological parameters.</p><p><b>CONCLUSION</b>Due to individual differences and immune pressure, HCV quasispecies can present with different patterns of evolution in the natural disease progression of chronic hepatitis C. HCV quasispecies evolution, due to its close correlation with the biochemical parameters, can be used to evaluate the severity and prognosis of chronic hepatitis C.</p>

Effects of Ellagic Acid on Melanogenesis and Melansome Transfer / 天津医药

Objective To investigate the effect of ellagic acid on human epidermal melanocyte melanogenesis and melanin transfer, and the mechanism thereof. Methods The human melanocytes and and keratinocytes were co-cultured and purified. After passing the second generation, cells of 1∶10 ratio were inoculated into the small dish (3 cm × 3 cm). The changes of melanin content and tyrosinase activity in melanocytes were detected before and after intervention with ellagic acid (100, 10 and 1 mg/L) for 48 h. The melanin transfer in cultured cells was detected by flow cytometry method. The 10 nmol/L arbutin was used as the positive control. Results The tyrosinase activity was down-regulated by ellagic acid in a dose-dependent manner. The ellagic acid can reduce the melanin content except for the 1 mg/L of ellagic acid. The melanin transfer was also inhibited by ellagic acid in a dose-dependence manner. Conclusion Ellagic acid can be used for skin-whitening cosmetic and the depigmenting effect might be due to the down-regulation of melanogenesis and melanin transfer.

Recombinant expression of hepatoma associated gene and its protein function / 中南大学学报(医学版)

OBJECTIVE@#To recombinant express hepatoma associated gene(HTA) and pre-test the function of HTA to determine the role of HTA in the development of liver cancer.@*METHODS@#HTA338-616 was amplified from HepG2 cells and cloned into the prokaryotic expression vector pET21a(+)-MBP. The proteins MBP and MBP-HTA were induced, purified by His-tag magnetic bead purification kit and identified by Western blot and ELISA. HepG2 cells were stimulated with MBP or MBP-HTA proteins. MTT assay and colony formation assay were employed to examine the proliferation of these cells and the changes of cell cycle distribution were determined by flow cytometry.@*RESULTS@#The prokaryotic expression plasmid pET21a(+)-MBP-HTA was successfully constructed. We got a 52 kD purified purpose protein.The proliferation of HepG2 cells stimulated with MBP-HTA was significantly higher than those stimulated with MBP and negative controls. HepG2 cells stimulated with MBP-HTA showed significant decrease fraction in G1 phase and increase fraction in S phase, and the cell proliferation was enhanced.@*CONCLUSION@#HTA protein can significantly promote the proliferation of HepG2 cells, which may be related to the promotion of G1 phase to S phase.

Expression and activity identification of a human nasopharyngeal carcinoma I50 anti-idiotype antibody / 中南大学学报(医学版)

Objective To obtain I50 anti-idiotype antibody and identify its activity in vitro.Methods I50 anti-idiotype (Id) antibody gene was amplified from the template of fuse 5-I50 by PCR to construct a prokaryotic expression vector pET25b-I50. The expression of pET25b-I50 in E. coli BL21(DE3) was induced by isopropylthio-β-D-galactopyranoside (IPTG) and was confirmed by SDS-PAGE and Western blot with Ab1(FC2) monoclonal antibody and an anti-hexahistidine tag antibody. The method of dialysis refolding was used to restore the activity of I50 anti-Id antibody, which was measured by Dot-ELISA and lymphocyte proliferation assay. Results The recombinant vector was successfully constructed and the recombinant protein was successfully expressed and purified with 90% purity. The relative molecular weight of the expressed protein was 15 kD, which was in accordance with expectation. The activity of I50 anti-Id antibody could be restored and could promote the proliferation of lymphocyte in a dose-dependent manner. Conclusion These results suggested that I50 anti-Id protein vaccine is likely an option in the therapy against nasopharyngeal carcinoma in vivo.

Dual effects of extract of Schisandra chinensis Baill on rat hepatic CYP3A / 药学学报

This study is to investigate the effects of aqueous extract of Schisandra chinensis Baill (WWZ), kadsurin, schisandrin A, schisandrin B and schisandrol B on rat hepatic CYP3A. Rats received a daily gavage of aqueous extract of WWZ for different times. The livers were harvested after gavage and subjected to microsome preparation. Microsomal CYP3A activity was determined by measuring the amount of the metabolite of testosterone (6 beta-hydroxytestosterone) with HPLC. Aqueous extract of WWZ, kadsurin and schisandrin A were incubated with microsomes obtained from rat. Microsomal CYP3A activity was determined by HPLC. Primary hepatocytes were separated and extracted from rat, then were treated with aqueous extract of WWZ, schisandrin A, schisandrin B and schisandrol B. Then, the expression of CYP3A1 mRNA was analyzed by RT-PCR. As for the in vivo assay, aqueous extract of WWZ significantly inhibited the enzyme activity of CYP3A after 12 h gavage. The inhibitory effect was converted to inductive effect after 3-day gavage. Aqueous extract of WWZ could induce the enzyme activity of CYP3A after 6-day gavage. Aqueous extract of WWZ and kadsurin showed a dose-dependent inhibition of CYP3A (IC50 of 487.8 microg mL(-1) and 6.2 micromol L(-1), separately). In rat primary hepatocytes, aqueous extract of WWZ (2.5 mg mL(-1)), schisandrin A (0.1 micromol L(-1)), schisandrin B (0.1 micromol L(-1)) and schisandrol B (10 micromol L(-1)) increased significantly the expression of CYP3A1 mRNA by 23%, 55%, 42% and 27%, respectively. Aqueous extract of WWZ could show dual effect on the enzyme activity of CYP3A in rat in vivo. Meanwhile, kadsurin showed a dose-dependent inhibition of the enzyme activity of hepatic CYP3A in vitro. And schisandrin A, schisandrin B and schisandrol B showed significant inductive effect on the expression of rat CYP3A1 mRNA.

In vitro anti-tumor immune mechanism of nasopharyngeal carcinoma bispecific anti-idiotype antibody / 中南大学学报(医学版)

Objective To compare the in vitro antitumor immune responses induced by bivalent bispecific anti-idiotype antibody G22-I50 and monovalent anti-idiotype antibody G22 and I50, and explore its possible mechanism. Methods Proteins G22-I50, G22, and I50 were induced and identified by Western blot and ELISA. Peripheral blood monoclear cells (PBMC) were isolated and stimulated with G22-I50, G22, and I50 anti-idiotype antibodies, respectively. MTT assay and LDH release test were employed to examine the proliferation and cytotoxicity of the PBMC. The levels of IFN-γ, IL-2, and IL-4 in the supernatant were detected by ELISA and changes of T lymphocyte subsets were determined by flow cytometry. Results Western blot showed that G22-I50, G22, and I50 had specific binding capabilities to FC2 (Ab1). The activities of G22-I50, G22, and I50 had recovered and these proteins could be used in the in vitro study. The proliferation and cytotoxicity of the PBMC stimulated with G22-I50 were significantly higher than those stimulated with G22 or I50, The level of IFN-γ and IL-2 in the culture supernatant of the PBMC stimulated with G22-I50 was higher than that in the G22 or I50 group, but the level of IL-4 did not increase.Compared with the G22 or I50 group, the proportion of CD4+ and CD8+ T cells and CD4+/CD8+ ratio significantly increased, and the proportion of CD4+CD25+ T cells significantly decreased in the PBMC stimulated with G22-I50. Conclusion G22-I50 has more potent immunogenicity and would enhance specific antitumor effect which might relate to improving PBMC proliferation, inducing the secretion of Th1 type cytokines, activating CD8+T cells, and suppressing the expression of CD4+CD25+ T cells.

Construction and expression of pDsRed-human apelin receptor recombinant plasmid in human embryo kidney 293 cells / 中国组织工程研究

BACKGROUND: Apelin/APJ system has a wide range of physiological functions,but its intracellular signal transduction,in particular,apelin receptor desensitization,internalization,resensitization degradation,have still no consistent opinion.OBJECTIVE: To construct eukaryotic expression vector expressing human apelin receptor(APJ)tagged to red fluorescent protein(pDsRED-express-C1),and to determine the expression in human embryo kidney 293 cells.METHODS: The plasmid pcDNA3.1-hAPJ was used as a template for PCR amplification of human APJ.Following PCR amplification the PCR product were removed and enzymatic digestion with EcoR I and BamH I.Same enzymes were used to cut vector pDsRED-express-C1.The digestive product was ligated by conventional methods of connection,then transfected into Competent E.coli TOP10.Single clones were picked plasmid extraction,followed by restriction enzyme digestion and finally DNA sequencing.The recombinant plasmid with correct sequencing was transfected into human embryonic kidney cells,PI staining,followed by the observation under a confocal microscope.RESULTS AND CONCLUSION: PCR amplified a 1.2-kb fragment,which was consistent with the expected size of the human APJ.The pDsRed-hAPJ recombinant plasmid was cut into two fragments,one corresponded to the pDsRED-express-C1 vector size,and the other fragment corresponded to APJ target fragment.Confocal microscopy analysis showed that,APJ was expressed mainly in the membrane of human embryo kidney 293 cells.The pDeRed-hAPJ eukaryotic plasmid expression vector was successfully constructed and effective expression of this fusion protein is achieved,which might be instrumental in the study of displacement and intracellular localization of human APJ.

The approach on methods of replacing tracheostomy tube after incision of trachea in 48 hours / 中国医师进修杂志

Objective To explore the more secure and available methods in replacing traeheostomy tube.Methods Sixty patients with incision of trachea were divided into group A(29 eases)and group B (31 cases)randomly.The new method was used in group A with putting a catheter into the old tracheostomy tube before taking it out,and placing the new tracheostomy tube under the guide of catheter.While the traditional method was used in group B.The heart rate,SpO2,manipulation time,perioperative haemorrhage, and the condition of entering the false passage were recorded.Results The heart rate,SpO2 were no significant difference before and after manipulation in group A,but the heart rate increased and SpO2 decreased in group B(P<0.01 or<0.05).The manipulation time was(50.5±4.2) s in group A,and (84.9±5.3) s in group B(P<0.05).The perioperative haemorrhage >3 ml and the condition of entering the false passage in group A(2 Cases,o cage)were less than those in group B(15 cases.5 cases)(P<0.01 or<0.05).Conclusion The new method in replacing tracheostomy tube which use an input catheter is more security and more availability.

Factors analysis on circadian rhythm of interdialytic blood pressure in maintenance hemodialysis patients / 中华肾脏病杂志

Objective To describe the circadian profiles of interdialytic blood pressure in maintenance hemodialysis patients and to investigate its related factors. Methods Ambulatory blood pressure monitoring (ABPM)was conducted in forty-four patients on regular hemodialysis during interdialytic days.Three groups were identified with their ambulatory blood pressure:non-hypertension group,controlled hypertension group and uncontrolled hypeaension group.Hemoglobin,creatinine,Kt/V,serum total calcium,serum phosphate,immunoreactive parathyroid hormone(iPTH),interdialytic weight gain and Morisky self-report scale etc.were assessed. Results (1)No significant difference was found in ABPM index ranged from decline in nocturnal systolic and diastolic blood pressure,AASI to dipper percentage in three groups.Nevertheless,24-h average pulse pressure in uncontrolled hypertension group was higher than the other groups[(80.06+13.41)vs(53.00±7.73),(57.85±21.97)mm Hg,all P＜0.01].(2)All the three groups showed the blood pressure profile of double-peak and trough.In the uncontrolled hypertension group,however,the nocturnal blood pressure decrease was not notable.(3)Nocturnal systolic blood pressure decline Was negatively correlated to iPTH(r=-0.039,P＜0.05).(4)In ten dippers out of all 44 patients,AASI was negatively correlated with nocturnal diastolic blood pressure decline (r=-0.748,P＜0.05). Conclusion Nocturnal blood pressure decline is correlated to iPTH and arterial selerosis.

Development and Application of Hospital Comprehensive Information System / 医疗卫生装备

Objective To provide guidance for patients and enhance their trust in the hospital by achieving openness and transparency in medical fees. Methods The modular structure of the three-tiered process was employed and various information machines, inquiry devices and doctors' workstations were connected to the Hospital Information System to form an integrated inquiry system of hospital-wide coverage. Results The Hospital Information System, with its versatile functions, easy operation, strong compatibility and good reliability, could provide patients with accurate information and more convenience. Conclusion The Hospital Information System can meet different needs of all patients.

Runx2 regulation and its isoforms / 中国药理学通报

Runx2 is a transcription factor essential for osteoblast differentiation and bone formation.Expression of Runx2 is initiated from two promoters,the distal P1 promoter and the proximal P2 promoter.These promoters give rise to two major protein isoforms with distinct amino termini,named as Runx2-TypeI and Runx2-TypeII.Here we review some integrated complex pathways(Wnt/LRP5/?-catenin,BMP/Smads/DPC4,Vitamin D/VDR/VDRE pathway,etc) and several key proteins(Msx2,Dlx5,Twists,etc) necessary for regulating Runx2 activity and bone formation,which give us new hints on controlling osteoblast differentiation by drug therapy and treating osteoporosis by gene therapy.