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Objective To construct Raji-Luc lymphoma cells with CD19 knockout using CRISPR/Cas9 technology and preliminarily validate their immune escape ability.Methods PB-CRISPR-CD19 small guide RNA(sgRNA)plasmids was constructed,the optimal sgRNA sequence was screened,and Raji-Luc cells with pCAG-PBase,PB-CD19 sgRNA,and PB-CRISPR-Cas9 were co-transfected.Stable knockout monoclonal cell lines were screened by flow sorting and limit dilution method and the knockout effect was verified through gene sequence testing.The expression of luciferase on the surface of the cell line was detected by microplate reader,CD19 CAR-T and CD38 CAR-T previously constructed in the laboratory were used as effector cells,and the immune escape ability of Raji-Luc CD19 KO cell line was verified by universal luciferase chemiluminescence method.Results The transfection efficiency of Raji-Luc CD19 KO cells prepared by electro transfection was high,and the knockout efficiency of the two monoclonal cells was more than 99%.There was no significant difference in luciferase expression compared to the original Raji-Luc cells,and CD19 CAR-T cells could not be activated to the kill them.Conclusion Successfully constructed Raji-Luc CD19 KO lymphoma cell line.
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Objective:To investigate the relationship between different obesity phenotypes and abnormal blood pressure in children and adolescents in Yinchuan city, and to provide appropriate treatment and intervention measures for obese children and adolescents.Methods:The current research design was adopted to facilitate the cluster sampling.A total of 1 047 children and adolescents aged 12 to 18 in Yinchuan were enrolled in this study from September 2017 to September 2018.There were 530 males and 517 females, with an average age of (13.93±1.24) years old.The questionnaire survey, physical examination and laboratory testing were carried out.Statistical analysis was performed by using SPSS 19.0 software.Results:Among the children and adolescents with normal weight, the composition ratio of the metabolically unhealthy normal-weight (MUNW) phenotype was 7.6%.In the obese cases, the composition ratio of the metabolically healthy obesity (MHO) phenotype was 20.2%.The blood pressure of MUNW [systolic pressure SBP: (119±13) mmHg(1 mmHg=0.133 kPa); diastolic pressure(DBP)(74±10) mmHg] and metabolically unhealthy obesity (MUO) [SBP (127±10) mmHg; DBP(74±7) mmHg] phenotypes were significantly higher than those of the metabolically healthy normal-weight (MHNW) phenotype (all P<0.05). The blood pressure of the MUO [SBP(127±10) mmHg; DBP(74±7) mmHg] phenotype was significantly higher than that of the MHO phenotype ( P<0.05). After adjusting for age, gender, and family history of hypertension, MUNW and MUO phenotypes were 5.93 (95% CI: 3.10-11.36) and 11.63 (95% CI: 6.37-21.24) times more likely to develop blood pressure abnormalities than MHNW phenotypes, respectively ( P<0.001). The MHO phenotype was 0.63 (95% CI: 0.08-4.93) times more likely to develop blood pressure abnormalities than the MHNW phenotype ( P=0.66). Conclusions:The MHO phenotype does not increase the risk of abnormal blood pressure, while the MUNW phenotype does.Therefore, it is recommended to identify the MHO phenotype and MUNW phenotype in order to provide appropriate obesity treatment and interventions for children and adolescents.
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Objective:To determine the seroprevalence of celiac disease in susceptible population, and to analyze the relationship between demographic characteristics, dietary habits, lifestyle and serological positivity so as to provide guidance for the prevention and treatment of celiac disease in Southern China.Methods:A total of 1 273 individuals who participated in Guangdong Province Health Screening Program in 2015, were selected as serologically positive subjects of celiac disease, including people with irritable bowel syndrome, colitis, diarrhea, anemia, low BMI, short stature, type 1 diabetes mellitus (T1DM), rheumatoid arthritis (RA), ankylosing spondylitis, psoriasis and bristol grade=6 or 7. All subjects were tested for serum IgA anti-tissue transglutaminase antibodies (TTGA), IgA antibodies against deamidated gliadin peptides(DGPA) and IgG against deamidated gliadin peptides (DGPG). Dietary habits, lifestyle and demographic characteristics were compared in subgroups.Results:The seroprevalence of celiac disease in susceptible population was 0.94% (95% CI 0.54%-1.64%) including 0.08% (1/1 273) for TTGA, 0.47% (6/1 273) for DGPA, and 0.39% (5/1 273) for DGPG. The seropositive rate was 3.6% (1/28) in patients with psoriasis, 2.1% (2/95) in the low BMI group, 1.9% (1/53) in T1DM group, 1.8% (3/169) in diarrhea group and 1.1% (5/463) in RA group. No significant difference was found in age, gender, high carbohydrate diet or lifestyle between the negative and the positive subjects. Conclusions:In Southern China, the seropositive rate of celiac disease is 0.94% in susceptible population, which prompts an urgent need of serological screening for early diagnosis.
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Objective:To evaluate the right ventricular(RV) function by two-dimensional speckle tracking echocardiography(2D-STE) in heart failure patients with different left ventricle diastolic dysfunction grade.Methods:Twenty-nine healthy volunteers(control group), sixteen heart failure patients with reduced ejection fraction(HFrEF group) and eighty-six heart failure patients with preserved ejection fraction(HFpEF group) were enrolled in the Second Affiliated Hospital of Chongqing Medical University from October 2018 to September 2019. HFpEF patients were further divided into four subgroups according to recommendations of the American Society of Echocardiography in 2016: normal left ventricular diastolic function group and left ventricular diastolic dysfunction groups with grade 1, 2, 3 (LVDD1, LVDD2, LVDD3). Conventional ultrasonic parameters were obtained in apical four-chamber images, including RV basal diameter(RVD1), RV middle diameter(RVD2) and RV longitudinal diameter(RVD3), RV end diastolic area(EDA), end systolic area(ESA) and fractional area change(FAC), peak systolic velocity(S′), myocardial performance index(MPI), tricuspid annular plane systolic excursion(TAPSE). 2D-STE was performed to observe strain parameters, including basal, middle, apical and the whole longitudinal peak systolic strain of RV free wall respectively (RVLSbas, RVLSmid, RVLSapi and RVLSfw). Then 2D strain parameters were compared among these groups, and the correlation between 2D strain parameters and conventional ultrasonic parameters were analyzed.Results:①TAPSE and S′ gradually decreased in normal group, HFpEF group and HFrEF group(all P<0.05). For the four subgroups of HFpEF, compared to LVDD3, TAPSE、S′ increased significantly in normal group, LVDD1 and LVDD2(all P<0.05). ②RVLSbas, RVLSmid, RVLSfw gradually increased in control group, HFpEF group and HFrEF group(all P<0.05). The same trend appeared in group LVDD1, LVDD2 and LVDD3(all P<0.05). ③In control group, HFpEF group and HFrEF group, RVLSfw showed strong correlation with LVEF( r=0.77, P<0.01) and middle correlation with age, TAPSE, and S′( r=0.41, -0.48, -0.40, P<0.01). In the subgroups of HFpEF, RVLSfw had middle correlation with LV diastolic function( r=0.59, P<0.01), and weak correlation with age( r=0.21, P<0.01), and the same correlations with B-type natriuretic peptide(BNP), FAC, TAPSE( r=0.28, -0.29, -0.29; all P<0.01). Conclusions:2D-STE can detect the early subtle impairment of RV systolic function in HFpEF patients.
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The incidence and mortality of heart failure with preserved ejection fraction (HFpEF) are increasing year by year. Right ventricle dysfunction (RVD) is an independent predictor of poor prognosis of HFpEF patients. Early evaluation of right heart structures and degrees of RVD are of great significance to the treatment and prognosis of HFpEF patients. The progresses of echocardiography in evaluation of the right heart structure and function in HFpEF were reviewed in this article.
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Objective This study aims to examine the association between leptin and postpartum depression symptoms. Methods Two hundred pregnant women were enrolled and their prenatal serum leptin levels were measured by using enzyme-linked immunosorbent assay (ELISA). The state of depression was first assessed on the third day after delivery and reassessed by a telephone follow up on the 42nd day using the Edinburg postnatal depression scale (EPDS) after delivery. Patients with postpartum depression symptoms with EPDS score ≥13 points after 3 days postpartum or 42 days postpartum were recruited as the depressive symptom group. Results The medians and lower and upper quartiles of the prenatal serum leptin levels in the depressive symptom group (the 3rd day) and the control group were 1.36 (1.15, 1.68) μg/L and 1.49 (1.28, 1.91) μg/L, respectively. After normal conversion, the serum leptin level was significantly lower in the depressive symptom group than in the control group (P=0.021). The medians and lower and upper quartiles of the prenatal serum leptin levels in the depressive symptom group (the 42nd day) and the control group were 1.17 (1.01, 1.36) μg/L and 1.50 (1.29, 1.90) μg/L, respectively. After normal conversion, the serum leptin level was significantly lower in the depressive symptom group than in the control group (P＜0.001). EPDS scores 3 days postpartum (r=-0.199, P=0.014) and 42 days postpartum (r=-0.254, P=0.002) were negatively correlated with prenatal serum leptin levels. Multiple logistic regression analysis showed that the prenatal leptin levels were associated with depressive symptoms at 42 days postpartum (OR=0.026, P=0.001). Conclusion Prenatal serum leptin levels may be associated with postpartum depressive symptoms, and high levels of leptin are protective factors for postpartum depression.
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Objective To evaluate the effectiveness of rapid hepatitis B vaccination with different vaccine dosages and types in adults.Methods Adults who were aged ≥20 years,negative in the detections of 5 HBV serum markers or only anti-HBc positive were selected from Chaoyang district of Beijing.They were divided into 4 community-based specific groups and given three doses of 10 μ g HepB-SCY vaccine,20 μ g HepB-SCY vaccine,20 μ g HepB-CHO vaccine and 10 μ g HepB-HPY vaccine respectively at month 0,1,and 2.Their blood samples were collected within 1-2 months after completing the three dose vaccination to test anti-HBs level by using chemiluminesent microparticle immunoassay.A face to face questionnaire survey was conducted,and x2 test,Mantel-Haensel x2 test,Kruskal-Wallis rank test and multiple logistic regression analysis were performed.Results A total of 1 772 participants completed vaccination and observation.Their average age was 48.5 years,and 62.75％ of them were females.The anti-HBs positive rates in the groups of 10 μg HepB-SCY,20 μg HepB-SCY,20 μg HepB-CHO and 10 μg HepB-HPY vaccines were 79.49％,84.34％,82.50％ and 74.15％,respectively (P=0.005),and the geometric mean titers (GMT) were 39.53 mIU/ml,62.37 mIU/ml,48.18 mIU/ml and 33.64 mIU/ml respectively (P=0.025).The overall anti-HBs positive rate and GMT were 79.01％ and 41.18 mIU/ml.The anti-HBs GMT of 4 groups declined with age.The differences in anti-HBs GMT among 4 groups minimized with age.The result of logistic modeling indicated that vaccine type and dosage,age and smoking were associated with anti-HBs statistically after controlling the variables of "only anti-HBc positive or not" and "history of hepatitis B vaccination".Conclusion Hepatitis B vaccination at dosage of 20 tg based on 0-1-2 month rapid schedule could achieved anti-HBs positive rates ＞80％ in middle aged and old people,which can be used as supplement of 0-1-6 month routine schedule.
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Objective To evaluate the effectiveness of rapid hepatitis B vaccination with different vaccine dosages and types in adults.Methods Adults who were aged ≥20 years,negative in the detections of 5 HBV serum markers or only anti-HBc positive were selected from Chaoyang district of Beijing.They were divided into 4 community-based specific groups and given three doses of 10 μ g HepB-SCY vaccine,20 μ g HepB-SCY vaccine,20 μ g HepB-CHO vaccine and 10 μ g HepB-HPY vaccine respectively at month 0,1,and 2.Their blood samples were collected within 1-2 months after completing the three dose vaccination to test anti-HBs level by using chemiluminesent microparticle immunoassay.A face to face questionnaire survey was conducted,and x2 test,Mantel-Haensel x2 test,Kruskal-Wallis rank test and multiple logistic regression analysis were performed.Results A total of 1 772 participants completed vaccination and observation.Their average age was 48.5 years,and 62.75％ of them were females.The anti-HBs positive rates in the groups of 10 μg HepB-SCY,20 μg HepB-SCY,20 μg HepB-CHO and 10 μg HepB-HPY vaccines were 79.49％,84.34％,82.50％ and 74.15％,respectively (P=0.005),and the geometric mean titers (GMT) were 39.53 mIU/ml,62.37 mIU/ml,48.18 mIU/ml and 33.64 mIU/ml respectively (P=0.025).The overall anti-HBs positive rate and GMT were 79.01％ and 41.18 mIU/ml.The anti-HBs GMT of 4 groups declined with age.The differences in anti-HBs GMT among 4 groups minimized with age.The result of logistic modeling indicated that vaccine type and dosage,age and smoking were associated with anti-HBs statistically after controlling the variables of "only anti-HBc positive or not" and "history of hepatitis B vaccination".Conclusion Hepatitis B vaccination at dosage of 20 tg based on 0-1-2 month rapid schedule could achieved anti-HBs positive rates ＞80％ in middle aged and old people,which can be used as supplement of 0-1-6 month routine schedule.
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Objective To optimize the producing area and parts of Gardenia roots. Methods Oleanolic acid 3-acetate was hydrolysed into Oleanolic acid in Gardenia roots from 10 different origins, and root, stem, leaf of Gardenia from Liuyang Hunan, and the content was determined by HPLC. Results The content of oleanolic acid 3-acetate in Gardenia roots of different origins from high to low was:Shaodong, Liuyang of Hunan>Anji of Zhejiang, Guiyang of Hunan>Ningxiang, Anhua of Hunan, Zhangshu of Jiangxi>Liling, Pinjiang, Youxian of Hunan. The content in root was 2 times of that in stem and leaf. Conclusion Experimental data were provided for the optimization of producing area and part of Gardenia roots.
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Objective To establish a method for efficient,accurate genotyping and nucleoside drug-resistant mutation analysis for hepatitis B virus ( HBV ).Methods The 48 HBV serum samples were collected from the Third People's Hospital of Taiyuan from July to August 2011,and HBV DNA were extracted using the commercial kit.The HBV whole genome and P gene were amplified and sequenced.Each HBV sample was genotyped by both constructing phylogenetic trees and genotyping software analysis.The results from two strategies were compared for every sample.Results A total of 48 HBV full genome sequences were identified into 12 B and 36 C genotype's by both constructing phylogenetic trees and genotyping software analysis,which was exactly the same as the analysis using P gene fragment sequencing.Seven forms of nucleoside drug-resistant mutation were found in the P gene for all the samples,with the ratio of 27.1％ ( 13/48 ),in which all the mutation forms were associated with lamivudine or adefovir,and no other nucleotide drugs-related resistance mutations existed.In addition,there were 11 B and 35 C genotype and 2 B/C hybrid type with the analysis using Real-time PCR genotyping for the 48 samples.Conclusion P gene sequencing can be used as a new clinical method for efficient,accurate HBV genotyping and resistant mutation analysis,which provides guidance for hepatitis B treatment.
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Ethanol is an attractive alternative to fossil fuels. Saccharomyces cerevisiae is the most important ethanol producer. However, in the process of industrial production of ethanol, both cell growth and fermentation of ethanologenic S. cerevisiae are dramatically affected by environmental stresses, such as thermal stress. In this study, we improved both the thermotolerance and fermentation performance of industrial ethanologenic S. cerevisiae by combined usage of chemical mutagenesis and genomic DNA mutagenesis-based genetic recombination method. The recombinant S. cerevisiae strain T44-2 could grow at 44 degrees C, 3 degrees C higher than that of the original strain CE6. The survival rate of T44-2 was 1.84 and 1.87-fold of that of CE6 when heat shock at 48 degrees C and 52 degrees C for 1 h respectively. At temperature higher than 37 degrees C, recombinant strain T44-2 always gave higher cell growth and ethanol production than those of strain CE6. Meanwhile, from 30 degrees C to 40 degrees C, recombinant strain T44-2 produces 91.2-83.8 g/L of ethanol from 200 g/L of glucose, which indicated that the recombinant strain T44-2 had both thermotolerance and broad thermal adaptability. The work offers a novel method, called genomic DNA mutagenesis-based genetic recombination, to improve the physiological functions of S. cerevisiae.
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Adaptación Fisiológica , ADN de Hongos , Genética , Etanol , Metabolismo , Fermentación , Calor , Microbiología Industrial , Métodos , Mutagénesis , Recombinación Genética , Saccharomyces cerevisiae , Genética , MetabolismoRESUMEN
Objective To establish the cell model of intractable epilepsy and to observe its neuronal damage and morphologic change of neurites.Methods The model was established by exposing hippocampal neurons to Mg2+ -free media for 3 hours on days 10 of culture.Expression of lactic acid dehydrogenase (LDH) in supernatant was measured as an index of neuronal damage.The morphologic change of neurons and neurites was observed by optical microscope and scanning electron microscope (SEM).Results Compared to the control group, level of LDH (U/L) was significantly increased in the model group at different time points (3 hours: 4.26 ± 1.28, 6 hours: 6.56 ±2.34 and 24 hours: 16.67 ±3.57, P <0.05).With time prolonging, release of LDH in the model group was gradually increased (F = 39.316,P <0.05).Under optical microscope, neurons of model group migrated closely to each other and neurite connections appeared to be gradually "reticulated" after Mg2+ -free media treatment for 24 hours; and the "reticulated" neurites connections become more obvious after 72 hours.Under SEM, neuronal membrane was rough and had several small depressions, neurites were interlaced in cluster.Conclusions Neuronal damage and morphologic change of neurites are verified in the cell model of intractable epilepsy.
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8 hours and without and binding nursing in these 3 groups respectively. Observed the condition of respiratory tract infection in 3 groups. Results The incidence rate of respiratory tract infection in binding nursing groups was 50%, which was significant higher than that of in no binding nursing group (P8 hours group was 66.67%, and this rate had an tendency of rising with the time of binding nursing lasting (P
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Objective To study the nursing method about percutaneous biliary tract stent implantation to cure malignant obstructive jaundice.Methods Careful perioperative nursing cares and finished post-hospital direction were applied among 21 patients with malignant obstructive jaundice when by percutaneous biliary tract stent implantation.Results There were 20 patients obtained successful operation,the successful rate was 95%.The postoperative of glutamic pyruvic transaminase were(96.60?89.36) U/L,the total bilirubin was(137.96?103.95) ?mmol/L,the directed bilirubin was((85.67)?62.95) ?mmol/L and the indirected bilirubin was(56.76?37.37) ?mmol/L.All the indexes which have mentioned above were significant lower than those of before operative,P
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0.05). (2) The serum leptin and TG concentration in breast cancer group were significantly higher , the HDL level was significantly lower than those in the groups of breast benign disease and healthy control (P0.05).(3) Multiple logistic regression analysis reveal statistically significant association between serum leptin, TG , ApoA1 and HDL-C levels and breast cancer incident.(ORLeptin=1.14, 95% confidence interval CI:1.076-1.210, ORTG =2.663, 95% CI:1.65-6.82; ORApoA1=5.726, 95%CI : 1.238 - 26.480 and ORHDL=0.035, 95% CI : 0.007 - 0.162, respectively). Conclusion:The increased serum leptin and ApoA1 levels and decreased HDL-C level may be the risk factors of breast cancer.
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Objective To evaluate the reliability and clinical application of measuring blood glucose and urea with electrodes and end reaction methods. Method Using the electrodes and end reaction methods, blood glucose and urea were measured respectively. The results and the relativity between the two methods were compared. Result Comparing the two methods for GLU and UREA, the correlative factors were 0.990 1 and 0.989 1 respectively. They had fairly good comparability. The linear ranges of their regression equations met the clinical needs. With favorable accuracy, the correlative factors were above 0.997 9. Conclusion The electrodes method was rapid and accurate, which was suitable for emergency tests. The ending reaction methods was fairly stable and has less interfering factors, which was fit for the routine use.
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Objective To evaluate the efficacy of balloon dilatation in the treatment of intrahepatic biliary strictures in patients with liver transplantation. Methods Of the 100 patients with liver transplantation, 16 patients had intrahepatic biliary strictures and received balloon dilatation treatment. Results Initial technical balloon dilatation was successful in 14 cases but failed in 2 cases. There were no procedure-related complications. 4 restenosis occurred and they were treated with repeated balloon dilatation treatment. Conclusion Balloon dilatation represented an effective and relatively safe treatment for biliary stricture in liver transplant recipients. For restenosis, balloon dilatation was also an effective treatment.
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Objective To explore the effects of Gantaikang on porcine serum-induced liver fibrosis in rats. Methods Hepatic fibrosis was induced in rats by intraperitoneal injection of 0.5ml porcine serum twice a week. Gantaikang was simultaneously administered intragastrically to rats in the intervention group at dose of 0.27g/kg?bw for 7 weeks. Then blood was collected, and serum levels of hyaluronic acid(HA), laminin(LN) and precollagen III(PCIII) were measured. Liver tissues was removed, and the pathologic changes of liver were observed by routine, Masson and James staining. The semi-quantitive analysis of collagen and reticulin in the liver tissues were performed. Results Gantaikang could effectively counteract the increase of HA, LN and PCIII level in serum, and attenuate hepatic collagen and reticulin proliferation induced by porcine serum in rats. Conclusion The results suggested that Gantaikang could, to a certain extent, prevent and attenuate immunologic hepatic fibrosis induced by porcine serum in rats.
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Objective To compare the result of T2/T3 transitional cell carcinoma (TCC) of the urinary bladder after segmental cystectomy, treated by postoperative radiation plus intravesical chemotherapy and postoperative intravesical chemotherapy alone.Methods From 1985 to Dec.1995 patients with T2/T3 TCC bladder cancer who had been treated by segmental cystectomy were eligible for this retrospective analysis. Fifty-eight patients received postoperative radiotherapy plus intravesical chemotherapy (RT+IVC) and 35 patients were given postoperative intravesical chemotherapy (IVC) with thio-TEPA or calmette-Gue'rin bacilli (BCG). For radiation,8 or 18 ?MV X-ray was given with total dose of 50-60 ?Gy. Vesicoclysis was performed on 50-60 mg thio-TEPA twice per week and 0.5 mg BCG per week.Results The 3-year local control rates of RT+IVC and IVC groups were 68.6% and 48.2% showing a difference statistically significant (? 2=4.08,P=0.044).The 3- and 5-year survival rates of RT+IVC and IVC groups were 70.7%,49.5% and 59.9%,35.7% ,showing no significant difference (? 2=1.77,P=0.184). Among the 5 year survivors of the RT+IVC patients, 78.6% had their bladder preserved. Though untoward radiation reactions were severer, they were tolerated well.Conclusions Combined radiation therapy plus intravesical chemotherapy is indicated for T2/T3 bladder cancer after segmental cystectomy. Multimodality therapy is more favored to improve both the local control and the possibility of preserving the bladder.
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Objective To evaluate the effect of radiotherapy(RT)combined with arterial infusion chemotherapy(AIC) in the treatment of locally advanced or recurrent rectal cancer. Methods From May.1994 to Dec.2000, 62 patients with locally advanced or recurrent rectal cancer were randomized into two groups: RT alone group(31 patients)and the combined group(RT+AIC, 31 patients). All patients were treated with conventional radiotherapy by 8 18 ?MV X ray to a total dose of D T40 50 ?Gy in 4 to 5 weeks for the resectable disease, or to a dose of 60 70 ?Gy in 6 to 7 weeks for the unresectable disease. However, the combined group received concurrent arterial infusion chemotherapy by DDP 70?mg/m 2 and 5 FU 600 ?mg/m 2 in 2 3 cycles. Results The response rates were 83.9% and 54.8% in combined group and RT alone group, respectively (P