Objective To explore the aging mechanisms of human adult adipose-derived stem cells (hADSCs) in vitro.Methods The hADSCs came from the production of normal patients with liposuction.The hADSCs were cultured in the 37℃ incubator and passed until the cells were senescent.The senescence-associated β-gal staining (SA-β-gal) and histone protein γH2AX staining were carried out in both young and old hADSCs.The quantitative real-time PCR (qRT-PCR) was conducted to check the difference of the transcription function of the young and old hADSCs.Results The cultured hADSCs would be the senescent cells when the PD number was more than 35.The aging cells became big and had large flat nuclei and much more lososomes in the cytoplasm.The SA-β-gal staining and γH2AX staining were mostly positive in the aging hADSCs (PD>35) compared with the self-renewing hADSCs (PD<20).The qRT-PCR results showed that the transcription levels of Nanog and Oct4 gene were very low in aging hADSCs compared with the self-renewing ones.Conclusions The hADSCs will be senescent (SEN) when cultured and passed in vitro for long time and the aging process is similar to that of stem cells in vivo.The aging model of hADSCs can be established by this method and will be very useful in the anti-aging drug, regeneration medicine, tissue engineering and stem cell therapy.

ObjectiveTo investigate the effect of focal ischemic preconditioning (IPC) on the expression of protein kinase-like endoplasmic reticulum kinase ( PERK ) and glucose-regulated protein 78 (GRP78) mRNA and protein after focal cerebral ischemia/reperfusion (I/R) in rats.MethodsAll 120 male SD rats were randomly divided into three groups: sham-operation group,middle cerebral artery occlusion (MCAO) group and brain ischemia preconditioning (BIP) group.Each group was further divided into 4 subgroups according to 12 h,1,2 and 3 d after I/R.The IPC models were made in order to measure the expression of PERK,GRP78 mRNA and protein by in situ hybridization and Western blot,and the apoptosis rate of neuron by flow cytometry. Results ①The expression of PERK mRNA increased and reached the peak at 12 h,then decreased continuously after 1 d.BIP could decrease its expression.The expression of PERK protein increased at 12 h and reached the peak at 24 h,then decreased continuously after 2 d.BIP could decrease its expression.②The expression both of GRP78 mRNA and its protein all increased and reached the peak at 12 h,then decreased continuously.BIP could increase their expression (mRNA:12 h: 136.70±9.53,F=32.265; 24 h:147.54 ±9.97,F=54.920; 2 d:158.16 ±9.44,F=45.374; 3d: 165.85±10.26,F=16.493,P＜0.05; protein:12 h: 1.319±0.116,F=5.619,P＜0.05; 24 h: 1.226±0.108,F=33.742,P＜0.01; 2 d:1.183 ±0.112,F =46.556,P ＜0.01; 3 d:1.115± 0.098,F =11.730,P＜0.05).③The rate of apoptosis neuron of rats in MCAO increased markedly at 12 h after reperfusion,and reached the peak at 1 d,then decreased continuously.BIP could decrease the rate of apoptosis neuron. Conclusion BIP can protect neurons through inhibiting the expression of PERK and inducing the expression of GRP78 after endoplasmic reticulum stress in rats.

This study was aimed to observe the effect of Bai-Zhu Fu-Ling (BZFL) Decoction in different proportion-ing on VIP and VIPR1 in Crohn's disease (CD) rats with spleen deficiency syndrome, in order to further explore the immunologic mechanism of BZFL Decoction on CD. The CD rat model with spleen deficiency syndrome was estab-lished using exhaustion and hunger. The model rats were treated by BZFL Decoction with different proportioning, and immunohistochemistry (IHC) was used to detect the expression of VIP and its receptor in colon tissues. The results showed that comparing to the blank control group, the level of VIP and its receptor of the model group significantly increased (P< 0.05). Comparing to the model group, the level of VIP and its receptor in BZFL Decoction B5 group (Rhizoma A tractylodis Macrocephalae:Poria = 12:15), B6 group (Rhizoma A tractylodis Macrocephalae:Poria = 15:12) and B7 group (Rhizoma A tractylodis Macrocephalae:Poria = 18:9) was significantly decreased (P< 0.05). It was con-cluded that the effect of BZFL Decoction of B5 group, B6 group and B7 group was better than other groups in VIP and its receptor which can regulate the VIP and its receptor, inhibit the releasing of inflammatory factors and reduce intestinal inflammation injury.

Objective To investigate the mechanism of sleeve gastrectomy combined with modified jejunoileal bypass (SG/MJIB) in improving the blood glucose of Goto-kakizaki (GK) rats.Methods According to the random meter method,the 28 GK rats were divided into 4 groups:sleeve gastrectomy with modified jejunoileal bypass (SG/MJIB),sleeve gastrectomy (SG),modified jejunoileal bypass (MJIB) and sham operation (SHAM).The changes of weight,food intake,fasting blood glucose,oral glucose tolerance test (OGTT),insulin tolerance test (ITT),plasma insulin and gastrointestinal hormonal (ghrelin and GLP-1) were monitored before and 16 weeks after surgery respectively.Results After surgery,the fasting blood glucose level in SG/MJIB group was obviously lower than that in SG,MJIB and SHAM groups (P＜0.05).From the 2nd week after surgery,OGTT of the SG/MJIB group was obviously improved compared to SG,MJIB,and SHAM groups (P＜0.02).In our post-op erative study,the insulin levels in SG/MJIB group were lower than those in SG,and SHAM groups (P＜0.05).Compared to MJIB and SHAM groups,ghrelin levels of SG/MJIB group were significantly decreased (P＜0.001),while GLP-1 levels of SG/MJIB group were higher than those of SG and SHAM groups (P＜0.01).Conclusions SG/ MJIB can improve insulin sensitivity and insulin secretion,and this effect is independent of the body weight and food intake.This study further validates that the gastrointestinal hormones play an important role in the pathogenesis and treatment mechanisms of type 2 diabetes mellitus.

Objective: To investigate lunch supply of public primary school canteens in Zhejiang Province, and to provide a basis for the scientific guidance of school lunch. Methods: During May to Jun. and Sept. to Oct. in 2019, lunch food supply was weighed and recorded and the number of diners in 44 public primary school canteens were summarized. Each investigation lasted for one week. Results: Food was mainly based on the taste of the students (40.91%) in the school canteen. About 45.45% of the schools referred to the Nutrition Guidelines of School Meals for students meals when making recipes in the school canteen. The supplies of cereals, vegetables, fruits, livestock and poultry meat, fish and shrimp, eggs, milk, soybean nuts, vegetable oil and salt were 109.05, 118.01, 0, 63.96, 9.25, 11.31, 0, 10.68, 10.47, 2.54 g. The supply of vegetable oil was basically the same as the recommended amount ( P >0.05). The supplies of energy, protein, calcium, iron, zinc, vitamin A, vitamin B 1, vitamin B 2, vitamin C, dietary fiber were 820.84 kcal, 32.79 g, 164.18 mg, 7.84 mg, 4.71 mg, 23.07 μgRAE, 0.41 mg, 0.35 mg, 20.47 mg, 2.34 g, 37.56% of energy from fat and 48.47% of energy from carbohydrate. The supply of vitamin B 1 was basically the same as the recommended amount ( P >0.05). There were no significant differences in all kinds of food and nutrients between urban and rural primary schools ( P >0.05). Conclusion Lunch supply is not optimistic in public primary school canteens in Zhejiang Province, with unreasonable structure and fails to agree with current nutritional recommendations. It is suggested that the scientific guidance of students meals should be carried out according to the survey results combined with the characteristics of local diet.

Objective:To investigate the value of logistic regression model based on the features of cone-beam breast CT (CBBCT) for the identification of benign and malignant masses in dense breast.Methods:The data of 106 patients (130 masses) with dense breast who underwent contrast-enhanced CBBCT examination and obtained pathological results from May 2011 to August 2020 were retrospectively analyzed as the training set. From August 2020, the data of 49 patients (54 masses) who met the same criteria were prospectively and consecutively collected and used as the validation set. Taking pathological results as the gold standard, the training set was divided into benign and malignant groups. The t-test, χ 2 test and Fisher′s exact test were used to compare the differences in CBBCT image characteristics between the two groups in the training set. A binary logistic regression model was established by multivariate analysis. ROC curves were used to assess the diagnostic efficacy of the model as a whole in the training and validation sets and the diagnostic efficacy of each feature in the model, and the cut-off value of the intensity (ΔCT) value was determined. The H-L method was used to test the goodness of fit of the model. Decision curve analysis (DCA) was drawn to validate the clinical power of the model. Results:Univariate analysis showed that the breast parenchymal background enhancement (BPE), shape, margin, lobulation, spiculation, density, calcifications, ΔCT value, enhancement pattern, non-mass enhancement, ipsilateral increased vascularity (IIV), and peripheral vascular signs had statistical difference between benign group and malignant group ( P<0.05). BPE, margin, ΔCT value and IIV were included in the multivariate analysis, the equation was logit( P′)=-8.510+0.830×BPE+0.822×margin+1.919× ΔCT+1.896 × IIV. The are a under curve of the model in the training set was 0.879 ( P<0.001) and in the validation set was 0.851 ( P=0.001). The are a under curve of BPE, margin, ΔCT value, and IIV in the diagnosis of malignant mass were 0.645, 0.711, 0.712, 0.775 (all P<0.05); the best cut-off value of ΔCT was 50.38 HU. The fit of this model was good ( P = 0.776). The DCA curve showed that when the risk threshold was 0.05-0.97, the net benefit rate was>0, and this model had some clinical value. Conclusion:The logistic regression model based on the features of CBBCT is helpful to distinguish benign and malignant masses in dense breasts.

The ultimate goal of synthetic biology is to build customized cells or organisms to meet specific industrial or medical needs. The most important part of the customized cell is a synthetic genome. Advanced genomic writing technologies are required to build such an artificial genome. Recently, the partially-completed synthetic yeast genome project represents a milestone in this field. In this mini review, we briefly introduce the techniques for de novo genome synthesis and genome editing. Furthermore, we summarize recent research progresses and highlight several applications in the synthetic genome field. Finally, we discuss current challenges and future prospects.
Animals ; CRISPR-Cas Systems ; Gene Editing ; methods ; Genetic Engineering ; methods ; Genome, Human ; High-Throughput Nucleotide Sequencing ; Humans