Objective To investigate the changes in biologic behavior of dendritic cells (DCs) after being modified by human telomerase reverse transcriptase (hTERT) gene. Methods In order to obtain the hTERT gene modified DCs, DCs were transfected with a replication-deficient recombinant adenovirus expression vector of hTERT. Then the expression to hTERT and PCNA was assessed by by Western blot, mature markers on DCs surface were detected by flow cytometry, and the proliferative capacity was determined by MTT method. Results Immunohistochemical staining and Western blotting showed that the expression of hTERT was upregulated obviously in DCs after being modified by hTERT gene. Flow cytometry indicated that the expression of CD83 and CD86 remained unchanged. The DC growth curve showed that the number of DC-hTERT was increased slightly in the first 2 weeks, meanwhile the number of DC/rAd-LacZ and DC was decreased obviously. Although the number in 3 groups was all decreased in the third week, the number of DC/rAd-hTERT was still greater than the other control groups. It was also found that the expression of PCNA was increased in DC/rAd-hTERT compared with that of immature DC, mature DC or DC/rAd-LacZ by Western blot. Conclusion After rAd-hTERT modification, life span of DC in vitro is extended and proliferative capacity is enhanced.

Objective The recombinant fluorescent eukaryotic expressing vector containing hTERT cDNA was transfected into human embryonic fibroblasts (hEFs) to explore the effects of exogenous hTERT on the type I and III collagens expression in hEFs. Methods p IRES2-EGFP-hTERT plasmid and pIRES2-EGFP plasmids were transfected into primary hEFs respectively by Lipofectin reagent. Expression of type I and III collagen was determined by Western blotting and the content of type I and III collagens in the cellular medium at 3 days after transfection were examined by radio-immunoassay. Results The expression levels of type I and III collagens in hTERT gene transfected hEFs(hEF-hTERT) were obviously higher than those in untransfected hEFs and vacant vector transfected hEFs(hEF-EGFP). The content of type I and III collagens in the cellular medium in hEF-hTERT cells at 3 days after transfection was also higher than that in untransfected hEFs and hEF-EGFP cells. Conclusions The synthesis ability of type I and III collagens in hEFs could be promoted by exogenous hTERT gene transfection.

In order to explore the effects of exogenous human telomerase reverse transcriptase (hTERT/hTRT/hEST2) on telomeric restriction fragment (TRF), telomerase activity and its subunits expression in human embryonic fibroblasts (hEFs), hTERT sense eukaryotic expression vector pIRES2 EGFP hTERT was constructed with DNA recombinant technique and then transfected into primary hEFs by Lipofectin method. TRF length, telomerase activity and changes in telomerase subunits expression were examined and evaluated in transfected and untransfected cells. The results showed that telomerase activity in pIRES2 EGFP hTERT transfected cells (hEF EGFP) was significantly higher than that in untransfected hEFs and vacant vector transfected cells (hEF EGFP) ( P

BACKGROUND: Following bone marrow mesenchymal stem cells (BMMSCs) infusion therapy, which factor promotes BMMSCs migrated to correct position is a key point, currently, adhesion molecule is thought to be playing an important role in mediating BMMSCs migration. OBJECTIVE: To investigate the expression of vascular cell adhesion molecule 1 (VCAM-1) and intercellular adhesion molecule 1 (ICAM-1) in rat BMMSCs. METHODS: BMMSCs were in vitro separated from rat bone marrow by directly adherence method. The expression of VCAM-1 and ICAM-1 were identified by using immunocytochemical staining, and the expression rates of antigen were tested by flow cytometry, in addition, their mRNA expressions were measured by RT-PCR. RESULTS AND CONCLUSION: Immunocytochemistry demonstrated that BMMSCs weakly expressed VCAM-1, but strong expressed ICAM-1. Flow cytometry showed that the expression rate of VCAM-1 was 6%, and the expression rate of ICAM-1 was 100%. RT-PCR showed that BMMSCs expressed a low level of VCAM-1 mRNA but a high level of ICAM-1 mRNA. It revealed under physiological condition, BMMSCs expressed a low level of VCAM-1, whereas they expressed a high level of ICAM-1.

Objective To investigate factors associated with prevalence, awareness, treatment and control of hypertension in Dagang Oil Field, and to provide the scientific evidence for the intervention measurements. Methods A cross-sectional, population-based survey was conducted in Dagang Oil Field from 2008 to 2010. Totally, 16 553 adults, 8 174 men (49.38%) and 8 379 women (50.62%), completed all of the questionnaires, measurement examination and blood specimen collection. Results The prevalence of hypertension was 27.30%. Multivariate logistic regression analysis showed that male, old age, overweight or obesity, family history of hypertension, marriage, education attainment, snoring, high heart rate, fasting blood glucose, serum total protein, and hyperuricemia were associated with hypertension. The awareness, treatment, control rates among all hypertensive subjects, and control rate among treated hypertensive participants were 52.65%, 40.07%, 15.51%, and 38.78%, respectively. Awareness was more common in those who were 45-64 years old , high medical costs, family history of hypertension, and smoking currently. Odds ratio (OR) (95% CI) of awareness was 5.24 (2.78 to 9.88), 24.11 (11.91 to 48.84) and 27.46(13.92 to 54.18) for medical costs 1-59.9 yuan/month, 60-199.9 yuan/month, and≥200 yuan/month, respectively. In contrast, exercising 2-4 times/week and longer sleep duration were associated with lower awareness. Medication treatment was more common in those who had old age, high medical costs, hypertension family history, coronary heart disease, prefer vegetable, while less common in those who exercise occasionally, sleep longer and drink alcohol currently. OR (95%CI) of treatment was 10.89 (3.76 to 31.56), 54.07 (19.20-152.24), and 87.31 (31.54 to 241.75) for medical costs 1-59.9 yuan/month, 60-199.9 yuan/month, and ≥200 yuan/month, respectively. Controlled hypertension was more common in those who had old age, high medical costs, exercise 2-4 times/week, coronary heart disease, prefer crude fibre diet, but less common in those who drink alcohol currently, have higher heart rate, and higher education attainment. OR (95%CI) of controlled hypertension was 13.59 (6.50-28.44), 14.67 (7.01-30.68), and 17.43 (8.34-36.39) for medical costs 1-59.9 yuan/m, 60-199.9 yuan/m, and≥200 yuan/m, respectively. Conclusion Despite high rate of awareness, treatment and control of hypertension still need to be strengthened.

Objective To observe the effects of calcium cyanamide (Rongbao) on the histological and morphological changes of Oncomelania hupensis snails in order to explore its molluscicidal mechanisms.Methods The serial snails' slides were fixed after soaking in a concentration of Rongbao leached liquor at different time.The histological and morphological changes of the snails were compared among these slides.Results After soaking in the Rongbao leached liquor for 48 h,the mantle epithelia,respiratory epithelia of the gill,liver cells,and muscle cells of gastropods were injured seriously,which resulted in the death of the snails directly.The death rates of the snails were 96.70% and 100% after soaking in the Rongbao leached liquor for 48 h and 72 h,respectively.Conclusion Rongbao is an effective molluscicide by damaging the several snail tissues.

Objective To investigate the effect of the replication-deficient adenovirus carrying miR-138(Ad-miR138) on the pro-liferation of human gastric cancer cell line BGC-823 and the possible mechanism .Methods The human gastric cancer cell line BGC-823 was infected with Ad-miR138 .Then the expression level of miR-138 was measured by RT-PCR .The growth curve of BGC-823 was measured using CCK-8 method .The ability of cell invasion was measured using Transwell chambers .Results After infected with Ad-miR138 ,the expression of miR-138 in BGC-823 cells was up -regulated significantly (1 .07 ± 0 .07 vs .4 .89 ± 0 .45 ,P<0 .05) .Absorbance of the 6th day decreased significantly (0 .52 ± 0 .06 vs .0 .77 ± 0 .06 ,P<0 .05) ,and the invasion ability was de-creased obviously (32 .00 ± 11 .00 vs .56 .00 ± 12 .00 ,P<0 .05) .Conclusion miR-138 can effectively suppress the proliferation and invasion of human gastric cancer cell line BGC-823 in vitro .

Ten novel podophyllotoxin derivatives (IIIa-IIIi and IV) were synthesized by three-step reactions using podophyllotoxin and N-benzyloxycarbonyl glycine-L-proline as raw material. The structures of the target compounds were confirmed by 1H NMR, 13C NMR and MS. MTT method was used to test anti-tumor activity of the target compounds on HepG2, THP-1, HeLa and MCF-7 cells. The results showed that all the target compounds had inhibitory activity against HepG2, THP-1, HeLa and MCF-7 cells, and the inhibitory activity of IIIa on HepG2 cells was the most prominent with an IC50 value of 0.58 nmol/L. The binding mode of compound IIIa and FAPα was studied by molecular docking. Compound IIIa could bind to multiple sites of FAPα enzyme.

OBJECTIVE: To develop a genotyping method for the Junior blood type and report on a rare blood type with Jr(a-). METHODS: Healthy O-type RhD+ volunteer donors of the Shenzhen Blood Center from January to May 2021 (n = 1 568) and a pedigree with difficult cross-matching (n = 3) were selected as the study subjects. Serological methods were used for proband's blood type identification, unexpected antibody identification, and antibody titer determination. Polymerase chain reaction-sequence specific primer (PCR-SSP) method was used for typing the proband's RhD gene. ABCG2 gene coding region sequencing and a PCR-SSP genotyping method were established for determining the genotypes of the proband and his family members and screening of Jra antigen-negative rare blood type among the 1 568 blood donors. RESULTS: The proband's ABO and RhD blood types were respectively determined as B and partial D (RHDDVI.3/RHD01N.01), Junior blood type Jra antigen was negative, and plasma had contained anti-D and anti-Jra. Sequencing of the ABCG2 gene revealed that the proband's genotype was ABGG201N.01/ABGG201N.01 [homozygous c.376C>T (p.Gln126X) variants], which is the most common Jr(a-) blood type allele in the Asian population. Screening of the voluntary blood donors has detected no Jr(a-) rare blood type. Statistical analysis of the heterozygotes suggested that the allelic frequency for ABCG2*01N.01 (c.376T) was 0.45%, and the frequency of Jr(a-) rare blood type with this molecular background was about 0.2‰. CONCLUSION A very rare case of partial DVI.3 type and Jr(a-) rare blood type has been identified. And a method for identifying the Junior blood type through sequencing the coding regions of the ABCG2 gene and PCR-SSP has been established.
Humans ; Blood Group Antigens/genetics* ; Genotype ; Genotyping Techniques ; Heterozygote ; Alleles ; Blood Donors ; Rh-Hr Blood-Group System/genetics*

OBJECTIVETo investigate genetic polymorphisms of 12 X chromosome short tandem repeat (X-STR) loci in ethnic Hebei Han population using an Investigator Argus X-12 amplification kit.

METHODSDNA was extracted for 198 unrelated individuals (96 males and 102 females) and amplified with a fluorescence labeled multiplex PCR system. PCR products were separated and genotyped with capillary array electrophoresis.

RESULTSOnly DXS10103 and DXS10101 showed significant linkage disequilibrium at the 12 X-STR loci. One hundred and forty-eight alleles, including 22 off-ladder (OL) alleles, were observed at the 12 X-STR loci in the population. The heterozygosity and polymorphic information content (PIC) were 0.5074-0.9143 and 0.4377-0.9079, respectively. The power of discrimination (PD) was 0.5074-0.9143 in males and 0.6876-0.9863 in females. The mean exclusion chance was 0.4377-0.9079 in the trios cases and 0.2984-0.8373 in the duo cases, respectively.

CONCLUSIONThe Investigator Argus X12 amplification system is highly polymorphic in ethnic Han population from Hebei and is useful for personal identification and paternity testing.

Alleles ; Asian Continental Ancestry Group ; ethnology ; genetics ; China ; Chromosomes, Human, X ; genetics ; Female ; Genetics, Population ; Humans ; Male ; Microsatellite Repeats ; Polymerase Chain Reaction ; Polymorphism, Genetic