Objective:To investigate the effects of myeloid-derived growth factor(MYDGF) on inflammatory response and osteoclast differentiation of RAW264.7 cells.Methods:The RAW264.7 osteoclast precursor cells were cultured and treated with different concentrations of recombinant MYDGF protein(rMYDGF), and their cell viability was assessed using the MTT assay. RAW264.7 cells were induced with lipopolysaccharide(LPS) to induce inflammation, and the expression of inflammatory mediators and cell polarization were observed after intervention with rMYDGF. The RAW264.7 cells were induced for osteoclast differentiation using receptor activator of nuclear factor-κB ligand(RANKL), and rMYDGF was added for intervention. Osteoclast differentiation was evaluated by tartrate-resistant acid phosphatase(TRAP) staining. The osteoclast resorption pits and the number of actin rings(F-actin rings) were observed under a microscope. Reverse transcription PCR was performed to detect the expression of activated T cell nuclear factor 1(Nfatc1), cathepsin K(CTSK), and c-Fos genes during osteoclast differentiation. The protein phosphorylation levels of nuclear factor-κB(NF-κB) signaling pathway proteins were detected using Western blotting.Results:MTT assay showed that rMYDGF did not significantly inhibit the viability of RAW264.7 cell when the concentration was lower than 100 ng/mL. Moreover, rMYDGF inhibited the expression levels of inflammatory factors and M1 cell polarization after LPS stimulation. Compared with the control group, the number and area of TRAP positive cells, the number and area of bone resorption pit were decreased in rMYDGF intervention group respectively, as well as the area of the F-actin ring was reduced and its shape was incomplete after rMYDGF intervention. Furthermore, rMYDGF reduced the expression levels of osteoclast-specific marker genes and inhibited the phosphorylation of NF-κB signaling pathway protein IκBα during osteoclast differentiation.Conclusion:MYDGF inhibits the inflammatory response and osteoclast differentiation of RAW264.7 cells.

A UHPLC-Q Exactive Orbitrap MS method was used to analyze the chemical constituents of the classical prescription Qianghuo Shengshi Standard Decoction(QHSS). UHPL conditions were as follows: Waters~(TM) UPLC~(TM) HSS T3 C_(18) column(2.1 mm×100 mm, 1.7 μm) and mobile phase of acetonitrile-0.1% formic acid aqueous solution. Mass spectrometry data of QHSS, each herb extract, and negative sample were collected in both positive and negative ion modes. The chemical constituents of QHSS were identified or tentatively identified based on the accurate molecular weight, retention time, MS fragmentation, comparison with reference substances, and literature reports. A total of 141 compounds were identified, including 18 amino acids, oligosaccharides, oligopeptides, and their derivatives, 19 phenolic acids, 44 coumarins, 18 flavonoids and chromones, 13 saponins, 17 phthalides, and 12 other components. This study comprehensively characterized the chemical constituents of QHSS, laying an experimental basis for the in-depth research on the material basis and quality control of QHSS.
Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal/chemistry* ; Gas Chromatography-Mass Spectrometry ; Mass Spectrometry ; Quality Control

Tea polyphenol （TP） is the main active constituent in tea， and epigallocatechin gallate （EGCG） is the main active constituent of TP that plays a biological role. Both of them can be used as immunomodulators in the treatment of tumor. In this paper， the anti-tumor mechanism of TP and EGCG was summarized from two aspects： enhancing immune effect and improving immune escape. The results showed that TP and EGCG could not only enhance the anti-tumor immune effect by promoting the activity of immune cells and the secretion of cytokines， but also improve tumor immune escape by regulating immunosuppressive cells， immune checkpoints， cytokines， immunomodulatory enzymes and JAK/STAT pathway， which has great potential in tumor immunotherapy.

Objective:To investigate the effect of alogliptin on bone loss in ovariectomized(OVX)mice.Methods:For animal experiments, thirty 8-week-old C57BL/6J female mice were divided into Sham group, OVX group, and OVX+ alogliptin group. OVX+ alogliptin group were administered with alogliptin in a dosage of 20 mg·kg -1·d -1 by gavage, Sham and OVX groups with equivalent saline. After 12 weeks intervention, serum bone anabolism indicators were detected, and Micro CT and HE staining were used to observe and analyze the bone trabecular structure of femur and tibia in mice. For in vitro experiments, bone marrow mesenchymal stem cells(BMSCs)were incubated with 100 μmol/L alogliptin for osteoblast differentiation. Alkaline phosphatase(ALP)and alizarin red S staining were used to determine the ALP activity and mineralization after osteogenic induction and culture. Real-time fluorescence quantitative PCR and Western blot were used to detect mRNA and protein expressions of osteoblast related genes. Results:Alogliptin intervention improved the biochemical indexes of bone anabolism and protected against bone microstructure deterioration to alleviate bone loss in OVX mice. Alogliptin stimulated osteoblast differentiation and elevated expression levels of Runt-related transcription factor 2(Runx2), ALP, osteocalcin, and osterix in in vitro experiments. Conclusion:Alogliptin can alleviate bone loss in OVX mice.

Objective:To observe the main clinical features and outcomes of type 2 diabetes mellitus patients after infection with COVID-19 and to compare them with those without diabetes mellitus.Methods:A single-center retrospective observational study was conducted in 88 in-patients who were diagnosed as COVID-19 from January 1 to February 26, 2020. They were divided into diabetic group and non-diabetic group, with 44 patients in each group. Patients′ medical history, laboratory examination, in-hospital treatment plan, and disease outcome were collected and compared.Results:The clinical symptoms of diabetic patients were varied, mainly fever(75.0%), cough(75.0%), fatigue(52.3%), and so on. The systolic blood pressure(SBP)was higher [131.50(120.00, 140.75) vs 125.00(120.00, 131.75)mmHg, 1 mmHg=0.133 kPa, P=0.021] and the oxygenation was lower [96.00%(94.25%, 97.00%) vs 97.00%(95.00%, 98.00%), P=0.038] at admission compared with the non-diabetic group. Hypertension and chronic kidney disease were more common in diabetic group. Fasting blood glucose [7.64(6.12, 15.43) vs 5.62(5.25, 6.50)mmol/L, P<0.01], interleukin-6(IL-6)[19.85(6.50, 43.38) vs 10.80(3.03, 20.90)pg/ml, P=0.046] in diabetic group were significantly higher than those in non-diabetic group. Secondary infection(27.3% vs 9.1%, P=0.027), ARDS(22.7% vs 4.5%, P=0.013)and shock(4.5% vs 0%, P<0.01)were more likely to occur in the diabetic group. More patients were treated with mechanical ventilation in the diabetic group(20.5% vs 4.5%, P=0.024). The diabetes group was more likely to progress to critical type(20.5% vs 4.5%, P=0.024), and the time to progress to critical state was shorter [3(1.75, 5.25) vs 6(3.00, 12.00)d, P=0.019). The duration of severe symptoms was longer in the diabetic group [26.5(15.00, 31.50) vs 9(8.00, 13.00)d, P=0.026]. Conclusion:The clinical symptoms of type 2 diabetes patients with COVID-19 are diverse. They are often combined with diseases such as hypertension and chronic kidney disease. The inflammatory reaction is more obvious and has more COVID-19 related complications and is more likely to progress into a persistent severe condition in a short time.

Objective To explore a newly discovered transmembrane protease serine 4 (TMPRSS4) isoforms and its molecular charcteristics in transfected colon cancer cells. Methods The named T4-1A and T4-1B of the TMPRSS4-isoforms were authenticated by the RT-PCR and Western blot,and then transfected to human colon cancer cells (DLD-1). Those stable transfected cells of migration and invasion were illustrated using wound healing assays and matrigel invasion assays. Results Successfully constructed T4-1A and T4-1B recombinant vectors,and obtained T4-1A and T4-1B transfected cell lines,and their T4-1A and T4-1B were highly expressed in stable DLD-1. Compared to cells transfected with empty vector of pcDNA6,the transfected DLD-1 with T4-1A enhanced migration and invasion were statistical significance (P < 0. 05). However,compared to pcDNA6 no significant1 difference was found for T4-1B. Moreover,the biological characteristics of T4-1A and TMPRSS4 were very similar. Conclusion The T4-1A and T4-1B is newly TMPRSS4-isoforms,and protease domain included to a T4-1A has further facilitated the migration and invasion of colon cancer cells,and further studies provide a theoretic base in the molecular biomedical characteristics of TMPRSS4.

Objective To assess the association of subclinical thyroid dysfunction with fractures. Methods Medline, Embase, Pubmed, Cochrane Library, CBM, CNKI, Wan Fang, and VIP databases were systematically searched from January 1990 to August 2015 to identify prospective cohort studies which have studied the risk of fracture in patients with subclinical thyroid dysfunction. The relative risks ( RR) of cohort studies were pooled respectively, depending on the result of heterogeneity test among the individual studies search. The Stata (version 13. 0) software was used for meta-analysis. Results Nine prospective cohort studies including 292460 participants were identified as eligible for the meta-analysis. RR of subclinical hyperthyroidism for fracture was 1. 39(95%CI 1. 24-1. 55);for hip fracture, RR was 1. 24(95%CI 1. 10-1. 40);for nonspine fracture, RR was 1. 32 (95%CI 1. 09-1. 60). Different gender for subclinical hyperthyroid was associated with higher fracture rates:for females, RR was 1. 15(95%CI 1. 04-1. 27); for males, RR was 1. 31 (95% CI 1. 08-1. 59). The incidence of fracture in patients with subclinical hyperthyroidism was higher during the follow-up. For subclinical hypothyroidism, the RR was 1. 21(95% CI 1. 03-1. 42). Subgroup analysis indicated that there were significant differences between endogenous/exogenous subclinical hyperthyroidism and euthyroid, but no differences between endogenous/exogenous subclinical hypothyroidism and euthyroid were found. Conclusion Subclinical hyperthyroidism is associated with an increased risk of fracture in the population, especially hip fracture and nonspine fracture. During the course of subclinical hyperthyroidism, the incidences of fracture should be noticed both in females and males. However, there is no evidence which could prove a definite association between subclinical hypothyroidism and the risk of fracture.

Objective To investigate the relationship between diameter of early gastric cancer as well as lymph node metastasis and expression of vascular endothelial growth factor(VEGF) and Matrix metalloproteinases-7(MMP-7)in gastric cancer. Method VEGF and MMP-7 expressions of gastric cancer tissue in 55 cases were examined by immumohistochemical assay. The diagnosis was confirmed by mucous stain and pathology. Results The level of VEGF and MMP-7 expression in group of early gastric cancer with diameter ≤ 10 mm was significantly lower than that of diameter ≥ 10 mm and ≤ 20 mm (P0.05). Conclusion It is significant that relation of the diameter of early gastric cancer with VEGF and MMP-7 expression, the cases of strongly positive over-expression of both VEGF and MMP-7 in tissue of patients with early gastric cancer are accompanied by more likelihood of lymph node metastasis.

Objective To explore the role of the autoantibodies against ?_1-adrenergic receptor(?_1-receptor)in the development of hypertension with renal failure.Methods The epitopes of the second extracellular loop of ?_1-receptor were synthesized and used respectively to screen sera autoantibodies from patients with hypertension with renal failure(61 cases),hypertension without renal failure(58 cases) and healthy blood donors(40 cases,control) by ELISA method.Results The positive rates of the autoantibodies ?_1-receptor(69%,42/61) in patients with hypertension with renal failure were higher than those of patients with hypertension without renal failure(19%,11/58) respectively(P

Objective To explore the roles of autoantibodies against ?1 adrenoceptor(?1-receptor)and M2 cholinergic receptor(M2-receptor)in patients with chronic renal insufficiency.Methods The epitopes of the second extracellular loop of ?1 receptor and M2 receptor were synthesized and used respectively to detect the sera autoantibodies against ?1 receptor and M2 receptor by enzyme linked immunosorbent assay(ELISA) in 76 patients with chronic renal insufficiency,60 cases with hypertension and 40 healthy controls.Results In patients with chronic renal insufficiency,the positive rates of the autoantibodies against ?1-receptor and M2-receptor were 56.7% and 38.1% respectively,which were much higher than those of patients with hypertension(18.3% and 11.7%) and higher than those of healthy controls(17.5% and 15.0%)(all P