Objective To establish the cell line stably expressing INSIG2 and observe its effecet on fat metabolism after overexpression of INSIG2.Methods The eukaryotic expression plasmid pcDNA3.1(+)-INSIG2 was constructed,which was transfected into 3T3-L1 cells.The expression of INSIG2 and related genes were detected by RT-PCR and immunohistochemistry,the contents of FFA in cell culture medium and adipocyte differentiation were detected by ELISA and Oil Red "O"staining respectively.Results After pcDNA3.1(+)-INSIG2 was transfected into the 3T3-L1 cells,the expression of INSIG1 mRNA and FAS mRNA were down-regulated,the content of FFA in the cell culture medium was decreased and adipocyte differentiation was drepressed.Conclusion The cell line stably expressing INSIG2 was successfully established,the transfected INSIG2 may have a drepressant effect on fat metabolism.

BACKGROUND:Adipose-derived stem cel s are totipotent stem cel s in the adipose tissue, and have the function of self-renewal and multi-directional differentiation. Human adipose-derived stem cel s are ideal seed cel s with stable genetic milieu and few rejections.
OBJECTIVE:To extract human adipose-derived stem cel s from human omental adipose tissue and to identify the cel s by adipogenic and osteogenic induction.
METHODS:Omental adipose tissues were col ected from surgical patients to isolate and culture adipose-derived stem cel s using type I col agenase digestion, filtration and centrifugation. Cel growth was observed and proliferative curve of human adipose-derived stem cel s were drawn by cel counting method to calculate the doubling time at logarithmic growth phase. After adipogenic and osteogenic induction, induced cel s were identified using oil red O and alizarin red staining, respectively.
RESULTS AND CONCLUSION:Human adipose-derived stem cel s were successful y isolated from the omentum tissues of surgical patients. Adherent cel s were fusiform-shaped and like fibroblasts. The growth curve of passage 3 cel s was in S shape, and the doubling time was 45.90 hours. After adipogenic and osteogenic induction for 2 and 3 hours, respectively, oil red O staining showed unequal-sized orange fat droplets, and alizarin red staining showed typical calcified nodules that were in orange. These findings indicate that adipose-derived stem cel s have the adipogenic and osteogenic capacity.

The ptrpose of this study was to investigate the effects of long-term estradiol and progesterone combined therapy on bone mineral density (BMD) in patients with Turner syndrome.Eight patients with Turner syndrome received estradiol and progesterone combined therapy for six years were observed and BMD was measured for each of them before hormone replacement therapy (HRT) and at an interval of one to two years after HRT and compared with that in normal age-sex matched women.BMD was significantly lower in patients with Turner syndrome than that in normal controls before HRT.All the patients with Turner syndrome had breast enlargement and irregular vaginal bleeding after HRT.BMD increased slightly in the patients with Turner syndrome after six-year HRT ,but still much lower than that in normal controls.Their BMD of the 2nd to 4th lumbar vertebra increased to (0.84±0.22) g/cm2 after HRT from (0.75±0.12)g/cm2 before it,with Z-score increased to -2.2±0.6 from -3.2±0.9,respectively; and overall BMD of the hip increased to (0.81±0.08) g/cm2 after HRT from (0.68±0.07) g/cm2 before it,with Z-score increased to-1.2 ± 0.3 from-2.2 ± 0.5,respectively.Long-term HRT can improve their BMD for patients with Turner syndrome but can not restore it to normal.

AIM: Some studies found that the glucose uptake ability of diabetes mellitus mice is stronger after feeding with taurine. Whether the taurine can affect adipocytes or not deserves further studies. This article investigates the effects of taurine on the differentiation of 3T3-L1 preadipocytes and detects the mechanics. METHODS: Experiments were performed in the Laboratory Center of Xiangya Third Hospital of Central South University from July to September 2006. ①3T3L1 preadipocytes were provided by Shanghai Cell Bank of China Academy of Sciences. ②3T3-L1 preadipocytes were cultured in high glucose DMEM medium containing fetal bovine serum of 0.10 volume fraction, 108 u/L benzylpenicillin and 80?1010 U/L streptaquaine. After confluence, cells at 5?107 L-1 were incubated in culture flask, and induced with 0.5 mmol/L IBMX, 0.5 mg/L insulin and 1 ?mol/L dexamethasone. The cells in an experimental group were intervened with taurine, whereas these in a control group were not treated with taurine. Oil O staining was used to observe the development of adipose cells. C3T3-L1 preadipocytes were treated with 10,20 mmol/L taurine respectively for 24 and 48 hours. RNA and protein were extracted in the control group and the experimental group. ③Genes related to adipose cells development was examined by RT-PCR and Western-blot. RESULTS: ①The number of cells stained by oil O was less in the experimental group than the control group in 3T3-L1 preadipocytes treated with 10 mmol/L taurine for 14 days. ②After C3T3-L1 preadipocytes were treated with 10,20 mmol/L taurine respectively for 24 and 48 hours, there were no changes in expression of Insig-2 protein, PPAR, Insing-2, adiponectin, adiponectin receptor, GLUT-4, AP-2 mRNA. CONCLUSION: Taurine inhibits the differentiation of 3T3-L1 preadipocytes, but the mechanics still need to be investigated.

Objective To study on NGF positive cells changes by the action of Danshen and explore the correlation between time and amount of NGF after traumatic brain injury (TBI)in rat.Methods72 rats were randomly divided into 3 groups,including sham group,brain injury and Danshen therapy group.TBI rat models were made by fluid percussion.NGF was determined on the 3rd,7th and 14th day after TBI.ResultsNGF positive cells after traumatic brain injury in brain reached the maximum level at the 3rd day(1.29±0.61),and it decreased since the 7th day(2.03±0.42),and it decreased to the lowest level on the 14th day (0.87±0.23).NGF positive cells after traumatic brain injury treated by Danshen were more than those patients who were treated without Danshen(1.77±0.54,3.35±0.37,0.65±0.21 vs 0.75±0.31,1.58±0.52,0.49±0.36,P＜0.05～0.01).ConclusionsThe expression of NGF was significantly increased in brain damaged tissues.This may be served as a useful marker of diagnosis in time-course of brain leison.Danshen injection can be used to protect brain after traumatic brain injury.

Objective: To investigate the health of gypsum miners in Hubei province and analyze the health effects of occupational disease hazards exposure on gypsum miners. Methods: In April 2017, occupational disease hazard factors were tested on the site of a gypsum mine, and 500 workers were selected by random sampling to conduct questionnaires, relevant data such as occupational health examination was collected, and descriptive statistical analysis was performed. Results: The main occupational hazards of gypsum miners were gypsum dust and noise. The time-weighted average concentration of 8 h in the workplace was 4.32 to 9.34 mg/m³, and the post pass rate was 69.2% (9/13) ; Respiratory dust 0.13-5.15 mg/m³, post pass rate 75.0% (3/4). Gypsum miners had finger dysfunction and muscle numbness, joint pain (29.2%, 88/301) and chest pain and breathing difficulties (17.6%, 53/301). Followed by tinnitus, auricle pus, running water (4.7%, 14/301), abnormal muscle tension (2.7%, 8/301). The exposure of occupational disease hazards was associated with respiratory, auditory and neurological symptoms of gypsum miners. Conclusion The long-term exposure of gypsum workers to gypsum dust, noise and other harmful factors may result in obvious symptoms of respiratory system and other health damage.

Humans ; Severe Acute Respiratory Syndrome ; diagnosis ; therapy

ObjectiveTo investigate the efficacy of Bushen Shengxue prescription and Yiqi Yangxue prescription in the treatment of chronic aplastic anemia and the effect on T cell subsets and the expression of T-box expressed in T cells （T-bet） and GATA binding protein 3 （GATA3）. MethodA total of 585 patients with chronic aplastic anemia who were treated in 19 hospitals in China from May 2018 to June 2021 were enrolled. With the prospective， double-blind and randomized control methods， the patients were randomized into three groups： kidney deficiency group， Qi and blood deficiency group， and control group. The three groups were respectively treated with Bushen Shengxue prescription granule， Yiqi Yangxue prescription granule， and Placebo （half the dose of Bushen Shengxue formula granules）. In addition， all of them were given oral cyclosporin and androgen. The treatment lasted 6 months， with 3 months as a course. The blood routine indexes， T cell subsets， and fusion genes T-bet and GATA3 before and after treatment were analyzed， and the safety indexes were monitored. ResultDuring the observation， a total of 75 cases dropped out and 18 were rejected. Finally， 161 cases in the kidney deficiency group， 164 in the Qi and blood deficiency group， and 167 in the control group were included. After 6 months of treatment， the total effective rate was 98.8% （159/161） in the kidney deficiency group， which was higher than the 79.9% （131/164） in the Qi and blood deficiency group （χ²=30.135， P<0.01） and the 61.7% （103/167） in the control group （χ²=70.126， P<0.01）. The total effective rate was higher in the Qi and blood deficiency group than in the control group （χ²=13.232， P<0.01）. After treatment， the hemoglobin （HGB） content increased significantly in three groups （P<0.05） as compared with that before treatment， particularly the kidney deficiency group （P<0.01）. After treatment， the white blood cell （WBC） count and platelet （PLT） count in the kidney deficiency group and the control group increased compared with those in the Qi and blood deficiency group （P<0.01）. There was no specific difference in neutrophils （ANC） after treatment among the three groups. At the same time point， the level of T helper type 1 （Th1） cells， Th1/Th2 ratio （P<0.05）， level of CD4⁺， and CD4⁺/CD8⁺ ratio （P<0.05） were significantly low in the kidney deficiency group among three groups. There was no significant difference in CD19^-， HLA/DR⁺， and CD25⁺ between the kidney deficiency group and the other two groups， but the T-bet of the kidney deficiency group and the control group was lower than that of the Qi and blood deficiency group （P<0.05）. ConclusionBushen Shengxue prescription exerts therapeutic effect on the aplastic anemia by improving the immunoregulatory mechanism， inhibiting the activity of immune system， modulating T cell subsets， suppressing Th1 and CD4⁺， and promoting bone marrow hematopoiesis. Moreover， it is safe with little side effects， which is worthy of further promotion.