To obtain single chain variable fragment (scFv) and bivalent single chain variable fragment (bsFv) against transferrin receptor, up-stream and down-stream primers were designed according to the complementary sequences of FR1 region of variable heavy (VH) and FR4 of variable light (VL), respectively, which contained inter-linker G4S and the restriction endonuclease SfiI, AscI and NotI. Two pieces of scFv fragments were first amplified through PCR and then inserted into plasmid pAB1, which could express scFv protein once induced by IPTG in the host bacteria. To express scFv and bsFv, E. coli TG1 was cultured in LB broth and was induced by IPTG. The restriction enzyme digestion map and DNA sequencing demonstrated that scFv and bsFv genes were successfully inserted into the expression plasmid. SDS-PAGE and Western blotting revealed the protein band at 35kD and 60kD, which were consistent with the molecular weight of scFv and bsFv respectively. Flow cytometry showed that scFv and bsFv harbored the specific binding activity with TfR expressed in various tumor cells, and the avidity of bsFv was higher than that of the parent scFv.
Base Sequence ; Cloning, Molecular ; Escherichia coli/genetics ; Escherichia coli/metabolism ; Genetic Vectors/genetics ; Hep G2 Cells ; K562 Cells ; Molecular Sequence Data ; Receptors, Transferrin/*immunology ; Recombinant Fusion Proteins/biosynthesis ; Recombinant Fusion Proteins/genetics ; Single-Chain Antibodies/*biosynthesis ; Single-Chain Antibodies/genetics

Objective To explore the feasibility of prediction of subacute infarct lesion volumes by processing ADC maps based on ADC cut-off values in patients with acute stroke.Methods MRI was performed in 20 patients with clinically diagnosed acute infarct less than 6 h after stroke onset.The MRI included a DWI and conventional MRI.The follow-up MR examinations of all the patients were carried out within 4-7 days after admission.The brain tissue, which showed abnormal high intensity both in DWI at the baseline and FLAIR at the reexamination , was regarded as the area of the initial ischemia core.Graphic-penumbra was regarded as the difference between initial DWI and follow-up FLAIR.The ADC values of the ischemia core, graphic-penumbra and the contralateral normal brain tissue , the relative ADC (rADC) were measured.The rADC value of the graphic-penumbra was defined as the ADC cut-off values.GE medical system based on ADC cut-off values was also tested in these patients to obtain ADC maps.The lesion volumes, the abnormal area seen on the DWI , ADC maps and follow-up FLAIR, were also measured.rADC values in different areas were analyzed by paired Student t test.Relationship between baseline DWI , ADC map and follow-up FLAIR was analyzed using Spearman rank-order correlation test , and Kruskal-Wallis H test was used to compare the volumes among three groups.Results Absolute ADC and rADC values gradually increased from the core to the periphery of the ischemic lesion.The absolute ADC values statistically differed from those on the contralateral side for both ischemia core and graphic-penumbra.The rADC values were significantly decreased in the ischemia core ( 0.620 ±0.116 ) compared with the graphic-penumbra values (0.809 ±0.097;t =8.083,P <0.01).The lesion volume of follow-up FLAIR [16.3 (4.7, 29.0) cm3 ] correlated highly with both volume predicted with the ADC map [13.5 (4.8, 25.4) cm3] (r=0.967, P<0.01) and baseline DWI volume [11.0 (4.4,30.4) cm3] (r=0.950, P<0.01).There was no difference between the volume of DWI , ADC map and follow-up FLAIR(H=0.168, P>0.05).Conclusions Data shows the high feasibility of prediction of subacute infarct lesion volumes by processing ADC maps based on ADC cut-off values in patients with acute stroke , without intravenous contrast material, and it provides a new method for outcome prediction.

OBJECTIVETo observe the effect of Yiqi Huoxue Qingre Huashi Recipe (YHQHR, a recipe capable of supplementing qi, activating blood, clearing heat, and dissipating dampness) on ulcer healing and Helicobacter pylori (Hp) eradication rate in Hp positive peptic ulcer patients, and to explore coccoid Hp occurrence in the eradication.

METHODSTotally 80 Hp positive peptic ulcer patients were assigned to the treatment group and the control groups by random digit table, 40 in each group. All patients received standard triple therapy of Western medicine for 2 successive weeks. Those in the control group additionally took omeprazole enteric coated tablet, 20 mg each time, once per day for 4 successive weeks. Those in the treatment group additionally took YHQHR, twice per day for 6 successive weeks. The ulcer healing was observed and recorded by gastroscope after discontinued medication of 14 days. The effective rate of ulcer healing under endoscope was statistically calculated. Rapid urease test (RUT) was performed in one small piece of tissue from corpora ventriculi and sinuses ventriculi using 14C breathe test (UBT). Gastric juice was collected from the stomach. Hp urease gene amplification test (urea A-PCR) was performed in living tissue from gastric antrum. Results obtained from the above three test methods were recorded and assessed to decide the final eradiation rate. Gastric mucosa tissue was observed under electron microscope,attempting to find non-eradicated Hp, which was further observed.

RESULTSThe total curative effect under gastroscope was 97.5% (39/40 cases) in the treatment group, obviously higher than that in the control group (80.0%, 32/40 cases) (P < 0.05). The eradication rate of Hp was 75.0% (30/40 cases), obviously better than that of the control group (52.5%, 21/40 cases) (P < 0.05). The total positive Hp numbers after treatment was 14C UBT (12), RUT (8), and urea A-PCR (27), respectively. The Hp positive rate detected by 14C UBT and RUT was lower than the Hp positive rate detected by urea A-PCR (P < 0.05). Rod-like and coccoid Hp bacteria could be observed under electron microscope.

CONCLUSIONYHQHR combined standard triple therapy was more effective than standard triple therapy alone in promoting ulcer healing and elevating the eradication rate of Hp.

Breath Tests ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Gastric Mucosa ; Helicobacter Infections ; drug therapy ; Helicobacter pylori ; Humans ; Omeprazole ; Peptic Ulcer ; drug therapy ; microbiology ; Urea

To understand the family sex education for young children in rural areas of Sichuan province and analyze the influencing factors. A multi-stage random sampling method was used to select 2246 parents of kindergarten children from rural areas in Sichuan province for a questionnaire-based survey.The Chi-square test and Logistic regression model were used for data analysis. It was found 1132(52.33%)parents had implemented family sex education for young children and 1031(47.67%)had not.Young children having asked sex-related questions(=1.536,95%=1.257-1.878),parents thinking that early childhood sex education is necessary(=3.691,95%=2.029-6.717),and parents having the intention to know early childhood sex education(=1.700,95%=1.274-2.269),and kindergarten having implemented early childhood sex education(=3.316,95%=2.515-4.372)were promoting factors for parents to conduct early childhood sex education,whereas a total annual household income at the middle level(=0.664,95%=0.456-0.968)was a hindering factor for parents to conduct early childhood sex education. Parents of young children in rural areas of Sichuan province have poor awareness of sex education,and the proportion of parents who have never conducted sex education for children is high.The total annual income of the family,whether the children have asked about sex-related questions,parents' attitude towards early childhood sex education,and whether the kindergarten has conducted the early childhood sex education are important factors that influence the level of children's family sex education.
Child ; China ; Humans ; Logistic Models ; Parents ; Sex Education ; Surveys and Questionnaires

Objective: To establish a prediction model for high-risk cardiovascular disease (CVD) among residents aged 35 to 75 years, so as to provide the basis for improving CVD prevention and control measures. Methods: Permanent residents aged 35 to 75 years were selected from Dongcheng District, Beijing Municipality using the stratified random sampling method from 2018 to 2023. Demographic information, lifestyle, waist circumference and blood biochemical indicators were collected through questionnaire surveys, physical examinations and laboratory tests. Influencing factors for high-risk CVD among residents aged 35 to 75 years were identified using a multivariable logistic regression model, and a prediction model for high-risk CVD was established. The predictive effect was evaluated using the receiver operating characteristic (ROC) curve. Results: A total of 6 968 individuals were surveyed, including 2 821 males (40.49%) and 4 147 females (59.51%), and had a mean age of (59.92±9.33) years. There were 1 155 high-risk CVD population, with a detection rate of 16.58%. Multivariable logistic regression analysis showed that gender, age, smoking, central obesity, systolic blood pressure, fasting blood glucose, triglyceride and low-density lipoprotein cholesterol were influencing factors for high-risk CVD among residents aged 35 to 75 years (all P<0.05). The area under the ROC curve of the established prediction model was 0.849 (95%CI: 0.834-0.863), with a sensitivity of 0.693 and a specificity of 0.863, indicating good discrimination. Conclusion The model constructed by eight factors including demographic characteristics, lifestyle and blood biochemical indicators has good predictive value for high-risk CVD among residents aged 35 to 75 years.

Objective To evaluate the influence and significance of preoperative design of surgical approach on stem cell transplantation via stereotactic surgery. Methods Six patients with stroke in the basal ganglia region were selected. The transplantation target and transcranial approach point were designed by magnetic resonance examination before stem cell transplantation via stereotacfic surgery to guarantee that the line connecting the transplantation target and transcranial approach point could avoid the important functional areas, the ventricular system and the softening focus. Postoperative magnetic resonance examination was performed to observe whether the practical target and surgical approach coincided with the preoperative design or not. Results The practical transplantation target was coincided with the designed transplantation target, distributed around the softening focus without implanted cells in the softening focus. Surgical approach was coincided with the preoperative design and it successfully avoided the important brain functional area, ventricular system and softening focus.Conelnsion The preoperative design of surgical approach can not only ensure the cells being exactly transplanted into the reservation target and guarantee the curative effect, but also promise the surgical approach successfully avoiding the important brain functional area, ventricular system and softening focus and reduce the operative injury.

Human infections with Lophomonas blattarum are rare. However, the majority of the infections occurred in China, 94.4% (136 cases) of all cases in the world. This infection is difficult to differentiate from other pulmonary infections with similar symptoms. Here we reported a case of L. blattarum infection confirmed by bronchoalveolar lavage fluid smear on the microscopic observations. The patient was a 21-year-old female college student. The previous case which occurred in Chongqing was 20 years ago. We briefly reviewed on this infection reported in the world during the recent 20 years. The epidemiological characteristics, possible diagnostic basis, and treatment of this disease is discussed in order to provide a better understanding of recognition, diagnosis, and treatment of L. blattarum infection.
Female ; Humans ; Lung Diseases, Parasitic/diagnosis/*parasitology ; Parabasalidea/*isolation & purification ; Protozoan Infections/*parasitology ; Young Adult

Objective:The aim of the current study is to understand the sleep disorders in children with autism and related factors.Methods:A case-control study method was conducted in this study which included 198 children autism who met the criteria of the Diagnostic and Statistical Manual of Mental Disorders,Fourth Edition (DSM-Ⅳ) and 233 cases of gender-and age-matched normal children.Their general demographic and sleep data were collected and the children with autism were assessed with Autism Behavior Checklist,Child Behavior Checklist and Autism Spectrum Screening Questionnaire.Correlation analysis and regression analysis were performed to compare sleep quality,sleep problems and the related factors of the two groups.Results:The scores of difficulty falling asleep,sleep restlessness,taking hold or shaking,restless sleep,interrupted sleep,nightmares,sleepwalking,night terrors,sleep snoring,nocturnal enuresis,sleep apnea,mouth breathing,sweating and sleep scores were higher in the autism group compared to the normal control group before falling asleep (P ＜ 0.05),while the proportion of daytime sleep time in the autism group was lower than the normal control group (P ＜0.05).With sleep status as dependent variable,linear regression analysis was conducted with children's birth weight,walking month age,speaking month age,father age,and autism spectrum as independent variables.The multiple linear regression analysis showed the scores of,sensory behaviors,body and object use behaviors,and self-help behaviors in Autism Behavior Checklist for autistic children were positively associated with their sleep (β =0.21,0.21,0.29,P ＜ 0.05).However,paternal age at birth in the autism group had negative associatin with the sleep of autistic children (β =-0.16,P ＜0.05).In the autism group,their total scores of Autism Behavior Checklist and the scores of language and social behavior in Autism Behavior Checklist were not associated with their sleep status score (P ＞ 0.05).Conclusion:It suggests that children with autism may have more sleep problems,and their sleep problems are related to autism symptoms including the scores of,sensory behaviors,body and object use behaviors,and self-help behaviors in Autism Behavior Checklist for autistic and their paternal age at birth.

2 in some patients with the loss of high and medium sized vWF multimers in plasma.Eight patients with vWD were identified, wherein two were characterized as type 1,4 as type 2A and 2 as type 3 respectively.Conclusion The panel of tests is suitable for diagnosis and classification of vWD.

BACKGROUNDIgA nephropathy (IgAN) is the most common primary glomerular disease. Transforming growth factor β1 (TGFβ1) plays an important role in pathogenesis of IgAN. Associations between the polymorphisms of TGFβ1 gene and the risk of IgAN remained inconsistent. A meta-analysis was conducted to investigate the association between polymorphisms in the TGFβ1 gene and IgAN susceptibility.

METHODSDatabases including Pubmed, EMBASE, ISI, et al. were searched to find relevant studies. Odds ratios (ORs) with 95% confidence intervals (CIs) were used to evaluate the strength of associations.

RESULTSTen studies involving 1770 cases and 1953 controls were included. Significant association between C509T polymorphism and IgAN risk was observed (OR 1.42, 95% CI 1.12-1.81, P = 0.0004; I(2) = 0%) in Caucasians by the overdominant model (CT vs. CC + TT), but no significant association was found (P = 0.200) in Asians by the dominant model (CC + CT vs. TT). Significant association between T869C polymorphism and IgAN susceptibility was found (OR 1.21, 95% CI 1.02-1.44, P = 0.030) in overall populations by the dominant model (TT + TC vs. CC). Subgroup analysis found T allele of T869C polymorphism was associated with IgAN susceptibility in Caucasians (P = 0.030), but not in Asians (P = 0.290).

CONCLUSIONBoth heterozygotes of C509T polymorphism and T allele of T869C polymorphism in TGFβ1 were associated with the risk of IgAN in Caucasians, but not in Asians.

Asian Continental Ancestry Group ; genetics ; European Continental Ancestry Group ; genetics ; Genetic Predisposition to Disease ; genetics ; Glomerulonephritis, IGA ; epidemiology ; genetics ; Humans ; Polymorphism, Genetic ; genetics ; Transforming Growth Factor beta1 ; genetics