In the context of precision medicine, although individual treatment of breast cancer under the guidance of molecular classi-fication has become the norm, a precision treatment program with increased efficiency and quality is still required. Compared with the traditional real-time fluorescent quantitative polymerase chain reaction (PCR), the droplet digital PCR (ddPCR) has obvious advantages in the detection of rare mutations and copy number variations, as well as the integration with the second-generation-sequencing tech-nology. This paper reviews the application of a ddPCR platform in different breast cancer subtypes and explores new horizons of breast cancer research through the ddPCR technology.

Purpose　The aim is to isolate and purify Protein C(PC) and Protein S(PS) from no-albumin human plasma by rivanol precipitation. Methods　The isolated and purified steps included adsorption onto and elution from barium, salting-out, ion-exchange chromatography, affinity chromatography,preparative isoelectric focusing and so on. Results　The molecular weights of the obtained PC and PS were (61±0.9)kD and (83±0.8)kD, respectively, the isoelectric point, 4.70±0.03 and 5.20±0.03,and the yield, 28.3% and 12.6%. The purified PC and PS were shown to be highly homogeneous by capillary zone-electrophoresis(CZE), and rich in Glu, Leu and Gly or Asp, Glu and Leu respectively. Conclusion　 The methods could be used for large-scale isolation and purification of PC and PS from no-albumin human plasma.

Objective:To systematically analyse the blood coagulation features of coronavirus disease 2019 (COVID-19) patients.Methods:An electronic search in PubMed, Scopus, Web of Science, EMbase, and CNKI to collect studies related to the blood coagulation features of COVID-19 patients from 1 January 2020 to 1 May 2020. Two reviewers independently screened literatures, extracted data and assessed the risk of bias of included studies. Then, the platelet count, D-dimer value, prothrombin time (PT), activated partial thromboplastin time (APTT) and fibrinogen of patients with different types of diseases were analyzed by using Stata12.0 software.Results:Thirty-nine retrospective studies involving 6 994 COVID-19 patients were included. The results of meta-analysis showed that:(1) compared with severe group, the platelet count (Weighted mean difference; WMD=20.11, 95% CI 11.53 to 28.69, P<0.001) and APTT (WMD=1.30, 95%CI 0.31 to 2.30, P=0.01) were found to be higher while D-dimer (WMD=-0.41, 95%CI-0.58 to-0.24, P<0.001), fibrinogen (WMD=-0.58, 95% CI-0.76 to-0.39, P<0.001) and PT (WMD=-0.51, 95%CI-0.92 to-0.10, P<0.001) were lower in mild group; the platelet count (WMD=-14.75, 95% CI-29.73 to-0.23, P=0.044) was found to be lower while D-dimer (WMD=1.06, 95% CI 0.65 to 1.47, P<0.001) was found to be higher in critical ill patients. (2)Compared with the survival group, the patients in death group displayed elevated levels of D-dimer (WMD=6.86, 95% CI 4.15 to 9.57, P<0.001) and PT (WMD=1.37, 95% CI 0.73 to 2.02, P<0.001) while platelet count (WMD=-36.40, 95% CI-63.23 to-9.58, P=0.008) remained low. Conclusion:Coagulation dysfunction was common in severe, critical ill and dead COVID-19 patients. Platelet count, D-dimer and PT levels were associated with the severity of the disease, and thus could be used as early warning indicators for the deterioration of the disease during hospitalization.

BACKGROUND: Irradiation therapy in the cancer patients after surgery may have negative effects on implant fixation. OBJECTIVE: To observe the effects of 60Co irradiation and trace element zinc on fixation strengths of titanium implant. METHODS: Thirty-six adult male rabbits were involved. One piece of titanium implant was separately placed into the bilateral proximal tibial heads in rabbits to set up animal models of titanium implants. The animals were randomly divided into control group, zinc supplement group, 60Co irradiation group and 60Co irradiation and zinc supplement group. Zinc sulfate of 10 g/L was administered intramuscularly to animals in the zinc supplement group at the dose of 4 mg/kg once per day, and those in the control group were treated with saline at the same dose. The animals in the 60Co irradiation group received 60Co irradiation at the dose of 45 Gy and saline by intramuscular injection. Those in the 60Co irradiation and zinc supplement group received 60Co irradiation and zinc sulfate. The animals were killed at 1, 4 and 12 weeks after treatment. The fixation strengths were measured and compared among groups. RESULTS AND CONCLUSION: (1) Zinc supplement made the titanium implants have higher fixation strengths, and reach a high level at 4 weeks after surgery. (2) 60Co irradiation significantly inhibited the fixation strengths of the implants. After 60Co radiation, zinc supplement stil could promote fixation strengths of titanium implants. (3) It is indicated that amount of zinc supplement after irradiation therapy can al eviate the negative effects of irradiation on implant fixation.

Objective:To investigate the effect of pilose antler polypeptide combined with Schwann cells modified by glial cell line-derived neurotrophic factor (GDNF)gene on the proliferation of human bone marrow mesenchymal stem cells (BMSCs)in vitro .Methods:According to the conventional method,the bone marrow (10 mL)was extracted from the healthy volunteers and was inoculated into the culture flask.The primary cultured cells were completely fused.The BMSCs were harvested at the 3rd generation and the cells were adjusted to 5 × 106 mL-1 . 4 μL GDNF gene modified Schwann cells was added into GDNF group,4 μL (10 mg· L-1 )PAP combined with GDNF gene modified Schwann cells was added into combination group,and only same amount of medium was added into control group.The proliferative activities,cell nuclear antigen (PCNA)levels and apoptotic rates of BMSCs in various groups were detected by enzyme-linked immunosorbent assay,ELISA method and Annexin Ⅴ-FIFC/PI cell apoptosis detection kit,respectively.Results:After primary culture for 48 h,most of the cells adhered to the wall,and the morphology of the cells changed into polygonal shape and few of them showed spindle.The passaged cells showed spindle spindle,the cells were confirmed as BMSCs,and all of them were non-hematopoietic stem cells.Compared with control group,the proliferative activities and the PCNA level of the BMSCs in GDNF group and combination group were increased (P <0.05)and the apoptotic rates were decreased (P <0.05);compared with GDNF group,the proliferative activity and the PCNA level of the BMSCs in combination group were increased (P < 0.05 )and the apoptotic rate was decreased (P < 0.05 ).Conclusion:PAP combined with Schwann cells modified by GDNF gene can promote the proliferation of human BMSCs in vitro .

The United States is the first country to implement DRG payment in the world, and its MS-DRG(medical severity DRG)version has been used for reference by many countries and regions. In order to ensure the universal applicability of DRG grouping scheme and adapt to the clinical reality, the MS-DRG grouping scheme should follow such grouping rules as similarity of resource consumption, clinical similarity and easy management of DRG groups. This paper presented the evolution of MS-DRG and expounded on its grouping rules in detail, for reference in the amendment and improvement of grouping rules in CHS-DRG.

Objective To investigate the expressions and clinical significances of breast cancer stem cell markers such as aldehyde dehydrogenase 1 (ALDH1) and transforming growth factor-β2 (TGF-β2) in patients with triple negative breast cancer.Methods Samples of 60 triple negative breast cancer tissues were investigated for the expressions of ALDH1 and TGF-β2 proteins by immunohistochemical staining.The correlation analysis,disease-free survival analysis and overall survival analysis were performed.Results The positive expressions of ALDH1 protein and TGF-β2 protein in the 60 breast cancer primary lesions were 23 cases (38.33％) and 38 cases (63.33％) respectively.The expression of ALDH1 protein was not correlated with tumor size (x2 =0.307,P =0.580),histological grade (x2 =4.244,P =0.120),clinical stage (x2 =0.982,P =0.612) or lymph node metastasis (x2 =1.111,P =0.292).The expression of TGF-β2 protein was not correlated with histological grade (x2 =4.651,P =0.098),lymph node metastasis (x2 =3.513,P =0.061),clinical stage (x2 =1.310,P =0.519) or tumor size (x2 =0.629,P =0.428).The disease-free survival time [(38.43±3.86) months vs.(53.38 ±2.58) months] and the overall survival time [(42.00±3.11) months vs.(53.84 ± 2.19) months] of ALDH1-positive patients were significantly shorter than those of ALDH1-negative patients,and the differences were statistically significant (x2 =8.490,P =0.004;x2 =11.270,P =0.001).The disease-free survival time [(42.81 ±3.32) months vs.(54.72 ±2.50) months] and the overall survival time [(44.74 ± 2.68) months vs.(57.18 ± 1.55) months] of TGF-β2 positive patients were significantly shorter than those of TGF-β2-negative patients,and the differences were statistically significant (x2 =4.300,P =0.038;x2 =8.900,P =0.003).The expression of ALDH1 protein was positively correlated with the expression of TGF-32 protein (r =0.360,P =0.005).Conclusion The ALDH1 phenotype is an independent predictor of poor prognosis.The activation of TGF-32 signaling pathway may be involved in the regulation of triple-negative breast cancer stem cells.

Osteosarcopenia is a geriatric disease in which sarcopenia and osteoporosis coexist. With the aging, the incidence of osteosarcopenia will increase significantly in the next few decades, with adverse consequences including a higher risk of falls, fractures, weakness, and death. Early diagnosis and intervention of osteosarcopenia are of great significance in improving the quality of life of the elderly. This article reviews the diagnosis, molecular mechanisms, adverse consequences, and possible treatment options for osteosarcopenia.