Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Mesenchymal Stromal Cells ; cytology ; Myocytes, Cardiac ; cytology

OBJECTIVETo assess the effect of Klotho gene transduction on the progression of hypertension and heart damage in spontaneous hypertensive rats (SHRs).

METHODSAn adeno-associated virus (AAV) carrying full-length mouse Klotho cDNA (rAAV.mKL) was constructed for in vivo expression of Klotho. Three different groups of male SHRs and a control group of sex and age-matched Sprague Dawley (SD) rats (5 rats per group) were used. The experimental groups of SHRs received an IV injection of phosphate buffered saline (PBS), rAAV.mKL and rAAV.EGFP, respectively. The control group only received equal-volume of PBS. The whole study has spanned 12 weeks. Plasma levels of insulin-like growth factor-1 (IGF-1) and insulin were measured with ELISA. The weight of whole heart was measured to calculate the heart weight index (HWI). EGFP expression of heart frozen sections was observed by fluorescence microscopy. Expression of mRNA and protein of Klotho, IGF-1, IGF-1 receptor (IGF-1R) and p-Akt were determined with RT-PCR, immunohistochemical analysis, and Western blotting. Hypertrophic myocardial cell and collagen fiber were observed by histological examination following Haematoxylin-Eosin and Masson staining.

RESULTSTransduction of rAAV.mKL can significantly prevent the increase of blood pressure in SHRs. Compared with the control group, the levels of Klotho mRNA and protein have both increased, and the plasma levels of IGF-1, insulin and glucose were elevated, whereas the expression of phosphor-Akt (also called Protein Kinase B, PKB) was decreased in the rAAV.mKL group. Furthermore, a decrease of hypertrophic myocardial cells and collagen fibers was noticed in the rAAV.mKL group compared with the control group.

CONCLUSIONThe Klotho gene can attenuate the progression of hypertension and abolishes myocardial hypertrophy and myocardial fibrosis. The protective effect observed in the rAAV.mKL group of SHRs may be attributed to increased Klotho protein and suppression of insulin and IGF-1 signaling pathways through inhibition of Akt phosphorylation.

Animals ; Blood Glucose ; Blood Pressure ; genetics ; Gene Expression ; Glucuronidase ; genetics ; metabolism ; Hypertension ; genetics ; metabolism ; pathology ; Insulin ; blood ; Insulin-Like Growth Factor I ; genetics ; metabolism ; Male ; Myocardium ; metabolism ; pathology ; ultrastructure ; Proto-Oncogene Proteins c-akt ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Inbred SHR ; Transduction, Genetic

To study the dynamic change law of bioactive constituents from Polygonum multiflorum, and to explore the optimal harvest period of P. multiflorum. Determination of stilhene glucoside, anthraquinones and catechin from P. multiflorum in different harvest times by MEKC-DAD, and principal component analysis (PCA) was used to comprehensive evaluation for bioactive constituents. There are obvious differences among the contents of active ingredients in various collecting periods samples, the content of stilbene glucoside was the highest in November, the total content of combined anthraquinone was the highest in November and December, the content of catechin was the highest in September. The comprehensive evaluation index obtained with principal component analysis showed that the sample collected in November is significantly higher than those with other samples. The optimal harvest period of P. multiflorum is November.
Electrophoresis ; Fallopia multiflora ; chemistry ; growth & development ; metabolism ; Time Factors

OBJECTIVETo study the relationship between resting heart rate and single nucleotide polymorphisms (SNP) at 3 sites of nitric oxide synthase (NOS) gene including NOS3 -922A/G, NOS3 894G/T and NOS2 -1173C/T SNPs.

METHODSGenomic DNA was gained from 211 Chinese Han nationality population. The SNPs of NOS3 -922A/G, NOS3 894G/T and NOS2 -1173C/T were genotyped by allele-specific primer-polymerase chain reaction (ASP-PCR) technique.

RESULTSThe distribution frequencies of GG, GT and TT genotypes of NOS3 894G/T and AA, AG and GG genotypes of NOS3 -922A/G and CC, CT and TT genotypes of NOS2 -1173C/T were consistent with Hardy-Weinberg equilibrium (P> 0.05). The resting heart rate of Chinese Han nationality population with AA genotypes was higher than that with GG genotype of NOS3 -922A/G (P< 0.01). The resting heart rate of the sub-population with GG genotype was higher than that with TT genotype of NOS3 894G/T (P< 0.05). There were no difference among the resting heart rates of the sub-populations with the allele genotypes of NOS2 -1173C/T.

CONCLUSIONThe resting heart rate of Chinese Han nationality population with mutated genotype GG of NOS3 -922A/G and with mutated genotype TT of NOS3 894G/T were lower than those with wild genotype of NOS3 -922A/G and NOS3 894G/T. The finding suggests that resting heart rate is associated with SNP of NOS3 -922A/G and NOS3 894G/T.

Adult ; Aged ; Aged, 80 and over ; Asian Continental Ancestry Group ; genetics ; China ; Female ; Gene Frequency ; Genotype ; Heart Rate ; Humans ; Male ; Middle Aged ; Nitric Oxide Synthase ; genetics ; Nitric Oxide Synthase Type II ; genetics ; Nitric Oxide Synthase Type III ; genetics ; Polymorphism, Single Nucleotide

Aim To explore the effect of Klotho (KL) gene transfection on the apoptosis of MC3T3-E1 osteo-blasts induced by dexamethasone(DEX). Methods MC3T3-E1 osteoblasts were transfected by recombinant adenovirus containing KL gene(Ad-KL) and recombi-nant adenovirus containing green fluorescent protein (GFP) gene(Ad-GFP). The apoptosis model was con-structed. The transfection efficiency of Ad-KL and Ad-GFP in cells were observed using inverted fluorescent microscope, and the level of KL mRNA and protein was detected by qPCR and Western blot,respectively. The cell viability after different concentrations of DEX acting on the cells and the viability of every research group were determined by cell counting kit-8 (CCK-8) assay. The apoptotic rate was evaluated by flow cytom-etry. The level of mRNA and protein was analyzed by qPCR and Western blot, respectively. The level of caspase-9 protein was detected by immunofluorens-cence assay. Results Cells were transfected by Ad-KL and Ad-GFP successfully. KL group and KL +DEX group had higher level of KL mRNA and protein than that in other groups. The optimum concentration of DEX was 2.0 mmol·L-1. When DEX acting on the cells, the cells viability decreased and apoptotic rate increased obviously in DEX group and GFP + DEX group. The level of Bax mRNA and protein presented a upward trend in DEX group and GFP +DEX group, while the level of Bcl-2 mRNA and protein was oppo-site. But after KL transfecting MC3T3-E1 osteoblasts, the markers described above in KL group had more dramatic improvement than in DEX group and KL +DEX group. Conclusions High-dosage DEX can in-duce the apoptosis of MC3T3-E1 osteoblasts, and the pro-apoptosis effect of high-dosage DEX in MC3T3-E1 osteoblasts can be suppressed by up-regulating KL gene expression level, suggesting that the glucocorticoid-in-duced osteoporosis might be improved by up-regulating KL gene expression level, and it may be a new target for the treatment of latrogenic osteoporosis induced by high-dosage glucocorticoid in clinic.

OBJECTIVETo evaluate the efficacy and clinical outcome of the treatment of thoracolumbar single compression fracture by using in situ rod rotation reduction and short segment pedicle screw at the fracture level.

METHODSFrom December 2008 to May 2010,12 cases of traumatic thoracolumbar single compression fracture (T11-L2) were treated, including 9 males and 3 females, with an average age of 35.8 years old (ranging from 24 to 52). There were 2 case with T11 fracture, 2 cases with T12, 6 cases with L1 and 2 cases with L2, without osteoporosis,pathological fractures or neurologic deficits. Radiographic data were collected preoperatively, 5 days postoperatively and at last follow-up (at least 12 months). Cobb's angle, vetebral compression ratio, internal fixation state were observed.

RESULTSAll patients were followed up from 12 to 30 months postoperatively,with an average of 19 months. There was no pseudoarticulation and solid bone fusion was achieved in all cases. There were no complications such as loosening or rupturing of internal fixation and so on. Sagittal kyphotic Cobb angle was corrected from preoperative (25.8 +/- 9.4) degrees to postoperative (6.7 +/- 2.3) degrees and (6.9 +/- 2.6) degrees at last follow-up. The percentage of vertebral compression was corrected from preoperative (42.5 +/- 10.4)% to postoperative (7.5 +/- 3.9)% and (8.4 +/- 4.5)% at last follow-up.There was significant difference between the postoperative data and preoperative data (P < 0.05), while the difference was not significant between the postoperative data and the last follow-up (P > 0.05).

CONCLUSIONThe thoracolumbar single compression fracture can obtain and maintain a good restoration by using the technic of in situ rod rotation reduction and short segment pedicle screw at the fracture level. The technique should be highly recommended.

Adult ; Female ; Fracture Fixation, Internal ; methods ; Fractures, Compression ; surgery ; Humans ; Lumbar Vertebrae ; injuries ; surgery ; Male ; Middle Aged ; Rotation ; Spinal Fractures ; surgery ; Spinal Fusion ; methods ; Thoracic Vertebrae ; injuries ; surgery

Objective:To explore the feasibility and clinical value of artificial intelligence-assisted breast ultrasound in screening breast cancer in Tibet.Methods:Two hundred and eighty-six women who participated in breast cancer screening in Shigatse People′s Hospital from August to September in 2021 were selected. The study included four groups. Group 1, ultrasound screening by senior breast ultrasound doctors from Shanghai; Group 2: local ultrasound doctors used intelligent-assisted ultrasound equipment for screening; Group 3: local ultrasound technicians used intelligent-assisted ultrasound equipment for screening; Group 4: ultrasound screening by local ultrasound doctors. The pathological results of screening positive cases and six-month ultrasound follow-up results of negative cases were set as the gold standard.Results:Twenty-seven lesions of 21 persons were screened positive. Pathology showed that 1 case of invasive ductal carcinoma, 1 case of severe atypical hyperplasia, 6 cases of fibroadenoma, 5 cases of breast disease, 14 cases of breast hyperplasia. Two hundred and sixty-five persons were screened negative, and the results of the six-month ultrasound follow-up were still negative. The accuracy, sensitivity, and specificity of group 2 were 0.966, 1, and 0.964 respectively; The accuracy, sensitivity, and specificity of group 3 were 0.935, 0.769, and 0.943 respectively; The accuracy, sensitivity, and specificity of group 4 were 0.860, 0.308 and 0.885 respectively. The accuracy and area under the curve of groups 2 and 3 were significantly different from that of group 4 (all P<0.001), and there was no significant difference from that of group 1 ( P=0.063, P=0.055). Conclusions:Artificial intelligence-assisted breast ultrasound screening technology can effectively improve the screening efficiency of non-breast ultrasound specialists and technicians. It is very suitable to solve the problems faced by grass-roots screening in Tibet and has great social significance and clinical value.

This work was designed to investigate proteins differentially expressed in cultivated Pseudostellaria heterophylla and its wild type using iTRAQ proteomics approach. The extracted proteins were digested using FASP method and identified by iTRAQ coupled with LC-MS/MS technology and then analyzed by Protein Pilot 5.0 search engine. Proteins differentially expressed were searched through comparison of relatively quantified proteins. The analysis was conducted using GO (gene ontology), KEGG and STRING. A total of 3 775 proteins were detected, among them, 3 676 proteins can be quantified, of which 127 proteins were up-regulated and 205 were down-regulated in cultivated Pseudostellaria heterophylla. We found 71 significantly differentially expressed proteins for further analysis. These proteins were classified into nine categories:heat shock proteins, transferases, oxidoreductases, lyases, isomerases, ligases, hydrolases, tubulin and translocases. The results indicated that the carbohydrate and cellular amino acids metabolism of cultivated Pseudostellaria heterophylla were weaker than its wild type and its ability of responding to stress was much stronger. GWD1, PHS1, GBE1, PGM, and BAM1 are the important proteins to regulate sucrose; metE and CYS are the key proteins that regulate amino acids in cultivated and wild Pseudostellaria heterophylla. This will provide the basic information for exploring the cause of secondary metabolites differences in different ecotype of Pseudostellaria heterophylla and the protein mechanism of its quality formation.

【Objective】 To explore the effects of perceived risk of COVID-19 of college students on their anxiety and depression, as well as the roles of attention to negative information and perceived social support, so as to provide theoretical basis for colleges and universities to formulate corresponding intervention measures. 【Methods】 By the convenience sampling method, totally 1 404 college students from Shaanxi and Henan provinces were investigated online by using General Information Questionnaire, Perceived Risk of COVID-19 Pandemic Scale, Attention to Negative Information Scale, Patient Health Questionnaire, Generalized Anxiety Disorder and Perceived Social Support Scale. SPSS 20.0 was used for data analysis, Pearson correlation method was used to explore the correlation between variables. The mediating effect of attention to negative information and the moderating effect of perceived social support were analyzed by PROCESS. 【Results】 The scores of anxiety and depression of the 1 404 college students included in the study were 4.03±4.48 and 6.21±5.41, respectively. The detection rate of anxiety symptom was 29.9%, and that of depression symptom was 44.4%. The risk perception of COVID-19 epidemic of the college students was positively correlated with attention to negative information (r=0.373, P<0.001), anxiety (r=0.227, P<0.001), and depression (r=0.226, P<0.001). Anxiety (r=0.553, P<0.001) and depression (r=0.497, P<0.001) were positively correlated with attention to negative information, while perceived social support was negatively correlated with the risk perception of the COVID-19 (r=-0.154, P<0.001), attention to negative information (r=-0.259, P<0.001), anxiety (r=-0.321, P<0.001) and depression (r=-0.278, P<0.001). The risk perception of COVID-19 affected the anxiety and depression of the students mainly through the mediating effect of attention to negative information. The total effect of risk perception of COVID-19 and anxiety was 0.227, and the mediating effect accounted for 80.18% of the total effect. The total effect of risk perception of COVID-19 and depression was 0.228, and the mediating effect accounted for 90.35% of the total effect. Perceived social support played a moderating role in the last half of this mediating model. 【Conclusion】 Risk perception of COVID-19 indirectly affects the occurrence of anxiety and depression in college students through attention to negative information, and perceived social support plays a moderating role in this mediating model. The findings suggest that when a risk event occurs, colleges and universities should pay attention to guiding students to adjust their attentional bias to external information, and give students enough care and support to improve their mental health.

Ambystoma mexicanum/immunology* ; Amputation ; Animals ; Biomarkers/metabolism* ; Blastomeres/immunology* ; Cell Lineage/immunology* ; Connective Tissue Cells/immunology* ; Epithelial Cells/immunology* ; Forelimb ; Gene Expression ; High-Throughput Nucleotide Sequencing ; Humans ; Immunity ; Peroxiredoxins/immunology* ; Regeneration/immunology* ; Regenerative Medicine/methods* ; Single-Cell Analysis/methods*