OBJECTIVE: To study the effect of overexpression of truncated type II TGF-beta receptor on transforming growth factor-beta 1(TGF-beta 1) autoproduction in normal dermal fibroblasts. METHODS: In vitro cultured dermal fibroblasts were treated with recombinant human TGF-beta 1(rhTGF-beta 1) (5 ng/ml) or recombinant adenovirus containing truncated type II TGF-beta receptor gene (50 pfu/cell). Their effects on regulating gene expression of TGF-beta 1 were observed with Northern blotting. RESULTS: rhTGF-beta 1 up-regulated the gene expression of TGF-beta 1 and type I procollagen. Overexpression of truncated receptor II down-regulated the gene expression of TGF-beta 1. CONCLUSION: Overexpression of the truncated TGF-beta receptor II decreases TGF-beta 1 autoproduction via blocking TGF-beta receptor signal. The results may provided a new strategy for scar gene therapy.
ObjectiveTo optimize HSP65-MUC1 fusion protein purification in pilot scale through protein purification techniques and identify the methods for biological activity detection. MethodsE. coli expressing HSP65-MUC1 was obtained by fermentation, then homogenized to obtain the supernatant. To acquire high-purity, high-quality HSP65-MUC1, the supernatant was treated with saturated ammonium sulfate, phenyl sepharose FF column and Q FF ion-exchange chromatography column purification. The expression of CD86 on the surface of DC cells treated with HSP65-MUC1 was determined with flow cytometry. ResultsE. coli containing pET28a-HSP65-MUC1 recombinant plasmid can effectively express target protein. A total of 413.7 mg of HSP65-MUC1 was obtained after 10 g of fermented cells was treated with saturated ammonium sulfate, phenyl sepharose FF column and Q FF ion-exchange chromatography column, and the purity was nearly 96%. Compared with negative control (10.13%±0.89%), purified HSP65-MUC1 could significantly improve the expression of CD86 on the surface of DC cells (29.98%±1.02%). ConclusionThe pilot scale production of purified HSP65-MUC1 has been effectively optimized, and the methods of its biological activity detection have been identified, which simultaneously provides the basis for clinical studies.
ObjectiveTo explore the effect of chronic unpredictable mild stress (CUMS) on the reproductive function of mice and provide a suitable animal model for reproduction and stress. MethodsA total of 240 female Kunming mice were feed for 5 days, and then divided randomly into the control group (n=90) and experimental group (n=150). The mice in the experimental group were stressed by 9 chronic mild unpredictable stress factors for 4 weeks and validated by open field test and sucrose consumption test. We administrated pregnant mare serum gonadotropin (PMSG)/human chorionic gonadotropin (HCG) for induction of superovulation and observed the ovarian response and embryo development potential. ResultsAfter 4-week CUMS stimulation, the weight gain, 2% sugar consumption test and open field test were significantly different between the mice in two groups (P>0.05). After PMSG/HCG was administrated, the antra follicles and preovulatory follicles significantly reduced significantly in the experiment group than that in the control group (P<0.05); the number of oocytes, fertilization rate, 2-cell embryos, D4 embryos, blastocysts, high quality embryo rate and D5 bed points were all significantly decreased in the experiment group than those in the control group (P<0.05). ConclusionThe CUMS female Kunming mice model is a kind of emotional stress animal model with low reproductive function, which is effective, operable and repeatable; it could be used for further study on the mechanism of reproductive medicine.
For treatment of pediatric inguinal hernia, we fabricated a device, i.e. so called "filling type pediatric hernia sac", which treats the problem from the abdominal cavity, through the abdominal and is a self-adaptive closer, using synthetic material. The device includes filling rack, self-adaptive umbrella support bar, bottom piece, outside pulling line and device fixing lines. The filling rack is composed of 2 concentric circles of 3.0 cm diameter with peripherally fixed together and can be pulled into the shapes of a ball or an olive. The supporting bar is structured of 3 pieces with 0.5 cm wide, 4.0 cm long, cross-fixed on top of the filling rack. The bottom piece is in a circular structure with a diameter of 3.0 cm, and it is connected to the filling rack bottom. Adjust positioning stay outside the fixed on the top of the device are connected at one end, and the other end free through filling the top frame connected with the bottom slice of central fixation. By using this device, we treated 37 pediatric inguinal hernia cases with 38 side-inguinal hernia successfully. The mean duration of post-operation follow-ups was 14.6±5.89 months, without hernia recurrence, obvious scar and hard sections of inguinal region. This device could provide a convenient, safe and effective plugging technology for children's pediatric hernia.
ObjectivesTo estimate the basic reproduction number of the novel coronavirus (2019-nCoV) and to provide support to epidemic preparedness and response.MethodsBased on the susceptible–exposed–infected–removed (SEIR) compartment model and the assumption that the infection cases with symptoms occurred before January 26, 2020 were resulted from free propagation without intervention, we estimated the basic reproduction number of 2019-nCoV according to the reported confirmed cases and suspected cases, as well as theoretical estimated number of infected cases by other research teams, together with some epidemiological determinants learned from the severe acute respiratory syndrome.ResultsThe basic reproduction number fall between 2.8 to 3.3 by using the real-time reports on the number of 2019-nCoV infected cases from People’s Daily in China, and fall between 3.2 and 3.9 on the basis of the predicted number of infected cases from international colleagues.ConclusionsThe early transmission capability of 2019-nCoV is close to or slightly higher than SARS. It is a controllable disease with moderate-high transmissibility. Timely and effective control measures are capable to quickly reduce further transmission.