Objective The aim of this study is to investigate the effects of the initial amount of total RNA input,adapter dilution and PCR amplification cycles on the results of small RNA library preparation and high-throughput sequencing.Methods Based on the sequencing reads number,the number of detected miRNAs and the accuracy of quantitative detection,we explored the effects of initial Total RNA,dilution ratio and PCR cycles on the quality of microRNA library preparation and high-throughput sequencing,respectively.Results For libraries preparation of the normal RNA input(1 μg),the adapter dilution combined with 22 PCR cycles could gain the best quality of sequencing.In the low-input libraries(10 ng),adapter dilution and increased PCR cycles would also improve the quality of sequencing,but as compared with the 1 μg library,there was lower correlation with microarray quantitative results in general.Conclusions The initial amount of RNAs input has the biggest effects on the quality of sequencing,and the quantity accuracy rate of low-input libraries is lower than the normal libraries generally.Increasing the PCR amplification cycles properly is indispensable to low-input libraries.

The fetal stage is a critical period for brain development. Intrauterine growth restriction(IUGR)af-fects the growth of fetus and impairs the brain development severely,resulting in long - term neurological sequelae such as motor and behavioral disorders. Since their brain underdevelopment originated in the uterus,postnatal interventions usually fail to improve the brain function. Then antenatal interventions become urgent to protect the brain,taurine has appeared a high practical value in improving the brain development. The adverse effects of IUGR on fetal brain develop-ment and the possibilities of antenatal interventions were introduced in this paper in order to improve the prognosis of the nervous system.

Objective To investigate the risk factors of early recovery in patients with ischemic stroke(IS).Methods 168 patients with IS received the conventional treatment and early rehabilitation were enrolled in the study.Single factor analysis was used to analyze the risk factors of the IS in the early treatment of rehabilitation in patients with and without success recovery rehabilitation success,and then application (unconditional Logistic regression analysis factors early independent risk factors for healing.Results Single factor analysis shows that age quartile (60years old),history of hypertension,coronary heart disease,diabetes,history,history of smoking history,anxiety and depression and successful rehabilitation negatively,life,education degree by law quartile (12 years) and mental work positively correlated with successful recovery.Many factors analysis shows that education degree by quartile (12)(OR=0.12,95％ 00.03 to 0.75),mental workers (OR=0.45,95％ CI 0.37 to 0.47) is the success of the recovery protection factor (P＜0.05),And age quartile (60) ( OR=2.12,95％ CI 1.21 to 3.21 ),anxiety ( OR =2.64,95％ CI 1.82 to 6.11 ) and depression (OR =1.45,95％ CI 1.13 to 1.96) is the success of the recovery independent risk factors (P ＜ 0.05 ).Conclusion Age quartile (60),anxiety and depression are IS successful independent risk factors for the recovery of the patients with early rehabilitation IS the risk factors should undertake the necessary intervention.

Objective The purpose of this study was to explore the infection characteristic of Toxoplasma gondii (TOX),Rubella virus (RV),Cytomegalovirus (CMV) and Herpes simplex virus Ⅱ type (HSV-Ⅱ) (TORCH) infection in neonate in Tianjin area.Methods TOX-IgM/IgG,RV-IgM/IgG,CMV-IgM/IgG and HSV-Ⅱ-IgM/IgG were detected in serum of 2 273 neonate during 2015～2016 with enzymelinked immunosorbent assay (ELISA).Results The positive rates of TOX IgM,RV-IgM,CMV-IgM and HSV-Ⅱ-IgM were 0.00％(0/2 273),0.00％(0/2 273),0.88％(20/2 273) and 0.00％(0/2 273),respectively and those of TOX-IgG,RV-IgG,CMV-IgG and HSV-Ⅱ-IgG were 3.65％ (83/2 273),86.45％ (1 965/2 273),95.82％(2 178/2 273) and 8.27％(188/2 273),respectively.There was 0.66％ percent (15/2 273) of examinees who were infected by none of TORCH pathogens.There existed significant statistical difference for positive rate between TOX-IgG,RV-IgG,CMV-IgG and HSV-Ⅱ-IgG (x2 =6.747,P =0.000) with consequence of the highest positive rate being CMV-IgG.The positive rates of TOX-IgG and CMV-IgM in neonate of 2016 were significantly less than those in 2015 (x2 =5.789～7.505,P=0.006～0.016) but that of HSV-Ⅱ-IgG of 2016 was statistically higher than that in 2015 (x2 =6.073,P =0.014).The positive rate of CMV-IgM in male neonate in 2015 was significantly higher than that in 2016 (x2 =5.054,P =0.025).As a whole the positive rates of TOX-IgG,RV-IgG,CMV-IgG and HSV-Ⅱ IgG had no differences between different years,so did those between gender groups (x2 =2.23～6.963,P=0.073～0.526).The positive rates of TOX-IgG,RV IgG,CMV-IgG and HSV-Ⅱ-IgG in female neonate in 2015 were statistically different from those in 2016 (x2 =8.247,P =0.041).The female neonate in 2015 had higher infection proportion of TOX-IgG compared with that in 2016 (x2 =6.992,P=0.008).TORCH infection detected in 2 273 cases of neonate had one pathogen infection and multi-pathogen infection with overall six patterns of TORCH infection and all infection patterns had no relationship with year and gender,respectively (P＞0.05).Conclusion RV and CMV were primary pathogens in TORCH infection for neonate in Tianjin area and there were recent infections by CMV.TORCH infection varied in different years and gender groups,which provided experimental data and basis for epidemiology and prevention of TORCH in neonate.

OBJECTIVETo observe the curative effect of Qianjin Kouchuang Jiawei Granule (QKJG) on recurrent oral ulceration [yin deficiency fire excess type (YDFET)].

METHODSTotally 120 patients who suffered from recurrent oral ulceration (YDFET) were randomly assigned to two groups, the experiment group and the control group, 60 in each group. Patients in the experiment group took QKJG, 20 g each time, twice per day, while those in the control group took Kouyanqing Granule (KG) , 20 g each time, twice per day. Fourteen days consisted of one therapeutic course, two for all. Scores for patients' symptoms and signs (ulcer area, exudation, hyperaemia, edema, the number of ulceration, burning sensation, and pain degrees) were assessed before treatment, at day 3 and 7 after treatment. Short-term efficacy was assessed by visual analogue scale (VAS). The total paralysis time and the total number of ulceration at month 12 after treatment were taken as judgment for long-term efficacy. Results Compared with before treatment in the same group, symptoms and signs were obviously improved at day 3 and 7 after treatment in the two groups (P < 0.05). Compared with the control group at day 3 after treatment, the improvement of edema, exudation, pain degree, and burning sensation was more obvious in the experiment group (P < 0.05, P < 0.01). The improvement of edema, pain degree, and burning sensation at day 7 after treatment was more obvious in the experiment group than in the control group (P < 0.05). As for short-term efficacy, the total effective rate was 86.67% (52/60 cases) in the experiment group and 83.33% (50/60 cases) in the control group, with no statistical difference between the two groups (P > 0.05). As for long-term efficacy, the total effective rate was 90.00% (54/60 cases) in the experiment group, significantly higher than that of the control group with statistical difference [81.67% (49/60 cases), P < 0.05]. At month 12 after treatment, the total number of ulceration was reduced and the paralysis time of ulcer attack prolonged in the experiment group, with statistical difference when compared with the control group (P < 0.05).

CONCLUSIONQKJG showed better long-term efficacy than that of KG in treating recurrent oral ulceration (YDFET).

Chronic Disease ; Drugs, Chinese Herbal ; administration & dosage ; therapeutic use ; Humans ; Pain Measurement ; Phytotherapy ; Treatment Outcome ; Yin Deficiency ; drug therapy

Objective To compare the transfection efficiency of galactosylated chitosan nanoparticle vehicle with chitosan nanoparticles vehicle,and observe the therapeutic effect of pGL3-hTERTp-TK on HCC cell line HepG2. Methods Preparing the chitosan and galaetosylated chitosan. Constructing the pGL3-hTERTp-TK plasmid and the Ch/DNA and GC/DNA complexes.Transfecting the HepG2 and the normal hepatic cell L-02 with chitosan/DNA and galactosylated chitosan/DNA complexs.Detecting the fluorescence and the expression of luciferase gene using the fluorescent microscope and the scintillation counter.Detecting the cell growth and apoptosis through the Caspase-3 and the flow cytometry. Results Two clear straps appeared in the agarose gel.The locations were 300 bp and 1100 bp.The relative lueiferase activity of pGL3-hTERTp-Luc+mediated by galactosylated chitosan was powerful than which of pGL3-hTERTp-Luc+mediated by chitosan in HepG2 by the scintillation counter.However,the relative luciferase activity was very weak in L-02.The same results were observed by fluorescent microscope.When the concertration of the GCV was 10 μg/ml(t=51.40,P=0.000),the HepG2 cell inhibition which was transfected by GC-pGL3-hTERTp-TK was obviously different from the L-02 cell inhibition which was transfected by GC-pGL3-hTERTp-TK in the statistics.The significant apoptotic rate was 65.28%in HepG2 which was transfected with GC/DNA,whereas it was only 10.80% in L-02. The significant apoptotic rate in HepG2 which was transfected with GC/DNA was very higher than the other groups (LSD, P ＜0.05). The significant apoptotic rate (10.80%) in L-02 which was transfected with GC/DNA was very higher than the group which was Ch-pGL3-control (LSD, P =0.000). The average fluorescence intensity of the HepG2 which is transfected by the Ch-pGL3-hTERTp-TK was 168.02± 3.68. The average fluorescence intensity of the HepG2 which is transfected by the GC-pGL3-hTERTp-TK was 204.45 ±3.45. The two groups had a significant difference in the statistics (t=-12.504, P＜0.05). Conclusion Galactosylated chitosan has higher transfection efficiency than chitosan. GC-pGL3-hTERTp-TK could specially attack HCC cell line and almost has no influence on normal hepatic cells.

Objective To investigate the intracellular and extracellular affinity between hepatitis B virus e antigen (HBeAg) and protein AK026018. Methods The gene AK026018 was amplified by reverse transcription polymerase chain reaction (RT-PCR) technique from HepG2 cell. The expressive vector of pGADT7-AK was constructed by routine molecular biological methods. The auxotroph yeast cells were cotransfected with pGADT7-AK and pGBKT7-eAg and plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing x-?-gal. To further prove the affinity between HBeAg and protein AK026018, translation was performed by using reticulocyte lysate, and immunoprecipitation was showed in vitro. Results The expressive vector was constructed and confirmed by DNA sequencing analysis and restriction enzyme digestion.The positive clones could grow on SD/-Trp-Leu-His-Ade/x-?-gal medium and were blue. The radioautographic band were found in immunoprecipitation analysis. Conclusions HBeAg could bind protein AK026018 in vivo and in vitro.

Objective To observe the effects of Lentinan(LNT) combined with Cisplatin (DDP) on the apoptosis induction in mice bearing Lewis lung carcinomas (LLC). Methods Lewis lung carcinoma cells (1?107) were subcutaneously inoculated into BALB/c mice. Mice were randomized into control groups and treatment groups. Observe the morphologic views under light microscope (LM) and electron microscope (EM). Results Compared with the control groups, the treatment groups were morphologic characteristic by relatively smaller in size, cell atypia were not evident, pathalogical karyokinesis were less interstitial blood vessel were fewer, while lymphocyte infiltrate into the interstitial. Apoptotic bodies could be seen under EM. Within all treatment groups, LNT+DDP group showed significant difference. Conclusions Lentinan combined with Cisplatin showed significant antitumor effects, and mechanism maybe inducing the tumor cell apoptosis and stimulate the proliferation of lymphocyte.

Objective To compare the value of four kinds of commonly used serological detection method for detecting Trepone-ma Pallidum,i.e.,Treponema Pallidum enzyme-linked immunosorbent assay(TP-ELISA),Treponema pallidum particle agglutina-tion test(TPPA),Treponema Pallidum rapid plasma reagin test(RPR)and Treponema Pallidum antibody detection(TP-AD,emul-sion method).Methods 5 870 specimens from outpatients and inpatients were screened by TP-ELISA.121 cases of detected posi-tive specimen were simultaneously detected by TP-AD,TPPA and RPR.Then the detection results were performed the comparative analysis.Results Among 5 870 specimens,121 cases were positive by ELISA,the detection rate was 2.06%.Among 121 positive cases,119 cases were positive by TPPA,the conformity degree was 98.34%,49 cases were positive by RPR,the conformity degree was 40.41%,113 cases were positive by TP-AD,the conformity degree was 93.38%.With the TPPA results as the standard,there was no statistically significant difference between TPPA and TP-AD(P >0.05),but there was statistically significant difference be-tween TPPA and RPR(P <0.01).Conclusion The four kinds of method have their applicability.ELISA d has good specificity and high sensitivity,and is simple to operate and suitable for the physical examination of large amount of pregnant women,parturients and normal people.TPPA has good specificity and high accuracy,is suitable for definite diagnosis.RPR is suitable for the monito-ring and the curative effect observation in the patients with the active stage of siphilis.Compared with ELISA,TP-AD has slightly less sensitivity,but good specificity and can be used for screening without specific instrument.