Objective To observe the effect of Tongxie Yaofang-containing serum on the cell cycle and apoptosis of colon cancer HCT116 cells,and to explore its possible biological mechanism.Methods Colon cancer HCT116 cells were divided into blank group and Tongxie Yaofang-containing serum group with different concentrations.Cell proliferation and activity detection-8(CCK8)was used to detect the effect of each group on the viability of HCT116 cells at 24,48 and 72 h;Flow cytometry was used to detect the apoptosis and cycle arrest of HCT116 cells induced by different concentrations of Tongxie Yaofang-containing serum;Western blot was used to detect the protein expression level of Cell Cycle Regulatory Protein P21(P21),Cyclin B1,cyclin-dependent protein kinases-1(CDK1),B-lymphoma-2(Bcl-2),Bcl-2-related X protein(Bax),phosphatidylinositol-3-kinase(PI3K),phosphorylated phosphatidylinositol-3-kinase(p-PI3K),protein kinase B(Akt)and phosphorylated protein kinase B(p-Akt)after different concentrations of Tongxie Yaofang-containing serum intervention.Results CCK8 showed that compared with the 10%blank group,10%Tongxie Yaofang-containing serum had a significant inhibitory effect on the viability of HCT116 cells only at 48 h(P<0.01);Compared with the 20%blank group,20%Tongxie Yaofang-containing serum could inhibit the viability of HCT116 cells at 48 and 72 h(P<0.01,P<0.05),and the increase of blank serum will not inhibit the viability of HCT116 cell,so 10%blank serum and 48 h were selected as the drug control and intervention time in the subsequent experiment.Flow Cytometry showed that,compared with the blank group,10%and 20%Tongxie Yaofang-containing serum could arrest HCT116 cell cycle in G2/M phase after 48 h of intervention(P<0.01),and induce HCT116 cell apoptosis.At the same time,Western blot showed that Tongxie Yaofang-containing serum could increase the expression of P21 and Bax to varying degrees,and reduces Cyclin B1,CDK1,Bcl-2,p-PI3K,p-Akt expression(P<0.05,P<0.01).Conclusion Tongxie Yaofang can inhibit the proliferation and induce apoptosis of colon cancer HCT116 cells,and its mechanism may be related to the inhibition of PI3K/Akt signaling pathway.

ObjectiveTo explore the effect of Tongxie Yaofang on the immune microenvironment of colorectal cancer in mice under chronic stress and the underlying mechanism. MethodA total of 40 male SPF BABL/C mice were randomized into normal group， stress group， Tongxie Yaofang group （13.65 g·kg^-1）， and Tongxie Yaofang-stress group （13.65 g·kg^-1）， with 10 in each group. Chronic restraint stress was induced in mice and administration （ig） of Tongxie Yaofang began after 7 days of stress. On the 14^th day， forced swim and tail suspension tests were used to examine the behavioral changes of mice after stress and the subcutaneous colorectal tumor was implanted in each group of mice. The effect of this prescription on the body mass and tumor volume of mice was observed. After the last administration， mouse serum and tumor samples were collected. The content of T lymphocytes （CD3⁺， CD4⁺， CD8⁺， and CD4⁺/CD8⁺） in tumor was detected by immunohistochemistry and flow cytometry and levels of corticosterone （CORT） in peripheral blood， and interleukin （IL）-2， interferon-γ （IFN-γ）， IL-6， and IL-10 in the serum were determined by enzyme-linked immunosorbent assay （ELISA）. The protein expression of inhibitor of nuclear factor-κB（IκB） kinase α/β （IKKα/β）， nuclear factor-κB （NF-κB）α （IκBα）， NF-κB p65， and phosphorylated （p）-NF-κB p65 was measured by Western blot. ResultCompared with the normal group， the stress group had large tumor volume （P<0.05）， low content of CD3⁺， CD4⁺， and CD4⁺/CD8⁺ （P<0.05， P<0.01）， high content of CD8⁺， low content of T helper 1 （Th1）-secreted IFN-γ （P<0.05）， high content of T helper 2 （Th2）-secreted IL-10 （P<0.05） and CORT （P<0.05）， high protein expression of p-NF-κB p65， NF-κB p65， and IKKα/β （P<0.05）， and low protein expression of IκBα （P<0.05）. Compared with the normal group， the Tongxie Yaofang group showed slow tumor growth， high content of CD3⁺， CD4⁺， and CD4⁺/CD8⁺ （P<0.01）， low content of CD8⁺ （P<0.05）， high content of Th1-secreted IL-2 and IFN-γ （P<0.05）， low content of Th2-secreted IL-6 and IL-10 （P<0.05）， low content of CORT， low protein expression of p-NF-κB p65， NF-κB p65， and IKKα/β （P<0.05）， and high protein expression of IκBα （P<0.01）. Tongxie Yaofang-stress group demonstrated slower tumor growth， higher content of CD3⁺， CD4⁺， and CD4⁺/CD8⁺ （P<0.01）， smaller content of CD8⁺ （P<0.05）， higher content of IL-2 and IFN-γ （P<0.05）， lower content of IL-6， IL-10 （P<0.05）， and CORT （P<0.05）， lower protein expression of p-NF-κB p65， NF-κB p65， and IKKα/β （P<0.05，P<0.01）， and higher protein expression of IκBα （P<0.01） than the stress group. ConclusionTongxie Yaofang can delay the growth of colorectal cancer under chronic stress and alleviate the deterioration of the immune microenvironment， possibly by inhibiting NF-κB signaling pathway， regulating the function of T lymphocyte subsets， and thus suppressing the secretion of pro-inflammatory factors.

Objective    To summarize our experience in the treatment of esophageal foreign bodies. Methods    A retrospective analysis of 149 patients of esophageal foreign bodies in the Second Affiliated Hospital of Air Force Military Medical University from December 2011 to May 2019 was carried out, including 75 (50.3%) females and 74 (49.7%) males with an average age of 57 (2-85) years. Results    There were 146 patients confirmed by endoscopy, and 3 patients were not found foreign body. Among the confirmed patients, 127 patients were removed by gastroscope and 19 patients were treated by operation. Esophageal foreign bodies are mainly related to the types of food. Jujube seed is the most common food foreign body in the northwest China. The injury rate of mucosal was 47.54% within 48 hours. The complication rate of taking out the foreign body after 48 hours was 100.0%. The success rate by endoscopy decreased (P=0.005), if the foreign body combined perforation. There was no statistical difference between the neck and other parts when using ultra-fine gastroscope (P=0.157). Conclusion    The sharper the foreign body is, the easier the perforation is. The earlier the foreign body is removed, the less complications are. The size of the foreign body determines the difficulty of endoscopic removal. Gastroscopy is the first choice for diagnosis and treatment, especially ultra-fine gastroscopy, and the foreign bodies that cannot be removed by endoscopy need surgical treatment.

A 75-year-old patient was admitted with "progressive dysuria for more than 2 months" in January 2017. The tPSA level was 498 ng/ml and then diagnosed as metastatic prostate cancer (cT 3bN xM 1). For the resistance of abiraterone, gene mutation was detected during the endocrine therapy. After 5 months of endocrine therapy, the serum tPSA was decreased to a minimum of 12.5 ng/ml. Since July, the serum PSA level gradually rebounded, and the endocrine therapy was altered to androgen deprivation therapy (ADT). The level of tPSA was maintained at 104 ng/ml in October 2017, and ADT was discontinued. After 1 year of discontinuation, the re-examination of PSA was 3 205 ng/ml. As the first-line regimen for mCRPC, abiratone and prednisone combined with goserelin was used. After 5 months of treatment, the level of tPSA still showed progression. The drug resistance of abiraterone was considered, so the treatment was discontinued. Next-generation sequencing technology (NGS) revealed the presence of AR, FGFR3 and RIT1 mutations, while no HRR germline mutation was detected. Docetaxel combined with ADT was performed. It was changed to comprehensive treatment of goserelin + docetaxel in March 2019. During chemotherapy CT images indicated significant reduction of pelvic lymph nodes and left inguinal lymph nodes, while bone metastasis showed stable condition. In April 2020, the chemotherapy was terminated for the lower extremity edema, joint pain and other related discomfort. The level of tPSA was 289 ng/ml after the last chemotherapy. DNA sequencing testing were performed again, and the mutation of AR and AR-V7 was negative. According to the results of genetic testing, the tPSA continued to decrease to 23 ng/ml after 6 months of abiraterone rechallenge, the imagings suggested that no disease progression. After AR mutation turns negative after chemotherapy, patients with refractory CRPC can still obtain a good PSA response such as tumor control and other clinical benefits from abiraterone. Abiraterone rechallenge is probably a new attempt for AR mutant patients with refractory CRPC.

A new coronavirus (SARS-CoV-2) has been identified as the etiologic agent for the COVID-19 outbreak. Currently, effective treatment options remain very limited for this disease; therefore, there is an urgent need to identify new anti-COVID-19 agents. In this study, we screened over 6,000 compounds that included approved drugs, drug candidates in clinical trials, and pharmacologically active compounds to identify leads that target the SARS-CoV-2 papain-like protease (PLpro). Together with main protease (M
Antiviral Agents/therapeutic use* ; Binding Sites ; COVID-19/virology* ; Coronavirus Papain-Like Proteases/metabolism* ; Crystallography, X-Ray ; Drug Evaluation, Preclinical ; Drug Repositioning ; High-Throughput Screening Assays/methods* ; Humans ; Imidazoles/therapeutic use* ; Inhibitory Concentration 50 ; Molecular Dynamics Simulation ; Mutagenesis, Site-Directed ; Naphthoquinones/therapeutic use* ; Protease Inhibitors/therapeutic use* ; Protein Structure, Tertiary ; Recombinant Proteins/isolation & purification* ; SARS-CoV-2/isolation & purification*

【Objective】 To verify the results of HBV DNA and HCV RNA screening under different brands of vacuum collection tubes for blood samples, storage temperature and storage time. 【Methods】 Experiment 1 was conducted as follows: blood samples were collected simultaneously from 52 voluntary blood donors using two brands(divided into group A and group B) of vacuum collection tubes for blood samples. The plasma separation of group A and group B were compared, and the effects of storage time on the NAT yield of HBV DNA and HCV RNA were statistically analyzed. Experiment 2 was conducted as follows: the effects of different storage temperature, time and tubes on the NAT yield of HBV DNA and HCV RNA samples with low viral load in group A and B were verified and compared in the simulated phlebotomy condition. 【Results】 In Experiment 1: After centrifugation, blood plasma layer and cells layer were separated completely in group A(100%, 52/52), but one sample was not well separated in group B(1/52, 1.92%). After 4 to 10 h after collection, blood samples of two groups were centrifuged and screened for HBV DNA, HCV RNA within 24 h. No positive samples were yielded and the Ct values of internal control(IC-DNA and IC-RNA) were uniform. In Experiment 2: Whole blood samples, stored for either 4 h or 6~10 h at 4 ℃ or 25℃ before centrifugation, showed no difference on the NAT-yield of HBV DNA nor HCV RNA samples with low viral load(P>0.05). Ct values of HBV DNA and HCV RNA of group A was similar to those of group B as centrifuged samples were stored for 24 h or 72~104 h at 4℃(P>0.05), but all increased as the storage time prolonged. Ct values of HBV DNA in group A increased from 33.45±0.29(24 h) to 33.82±0.08(72~104 h) and HCV RNA from 35.21±0.20 to 36.12±0.43; HBV DNA from 33.46±0.25 to 34.30±0.60 and HCV RNA from 35.47±0.24 to 36.49±0.51 in group B. 【Conclusion】 Under certain laboratory condition, different storage time, storage temperature and tubes shed few effect on the NAT-yield of HBV DNA and HCV RNA samples with low virus loads. However, it is suggested that the blood sample be detected within 72 h after centrifugation at 4 ℃ storage.

BACKGROUND: Electromagnetic navigation bronchoscopy is a tool that can accurately navigation peripheral lung lesions. Because of electromagnetic navigation bronchoscopy (ENB) is too expensive, it has not been widely used in China. It is urgent for us to summarize experience in clinical application, especially in the diagnosis of pulmonary nodules. METHODS: The clinical data of patients with pulmonary peripheral lesions (PPLs) in our department undergoing ENB biopsy between July 2017 and December 2018 were retrospectively analyzed. RESULTS: There were 18 patients with 21 PPLs (10 males and 8 females). Among them, 11 patients got the final pathological diagnosis, 8 cases were diagnosed with adenocarcinoma lung cancer, 1 case was diagnosed with tuberculosis and 2 cases were diagnosed with small cell lung cancer. The positive rate of diagnosis was 61.1%. The sensitivity was 73.3%. The positive diagnosis rate is related to the size of the lesion, the positive diagnosis rate for lesions >2 cm is 100.0% (P=0.04). CONCLUSIONS Electromagnetic navigation bronchoscope is safe and effective in clinic. It has a high positive rate for the diagnosis of peripheral lung lesions larger than 2 cm, ENB has broad clinical application prospects.

Objective    In this study, three-dimensional printed (3DP) titanium implants were used for skeletal reconstructions after wide excision of chest wall. 3DP titanium implants were expected to provide a valid option with perfect anatomic fitting and personalized design in chest wall reconstruction. Methods    There were 13 patients [mean age of 46 (24-78) years with 9 males and 4 females] who underwent adequate radical wide excision for tumors and chest wall reconstruction using 3DP titanium implants. Surgical data including patient demographic characteristics, perioperative clinical data and data from 1-year follow-up were collected and analyzed. Results    Six patients of rib tumors, six patients of sternal tumors and one patient of sternal pyogenic osteomyelitis were finally selected for the study. The chest wall defect area was 221.0±206.0 cm2. All patients were able to maintain the integrity of the chest wall after surgery, and no abnormal breathing was found, achieving personalized and anatomical repair. Thirteen patients were successfully discharged from the hospital. Two patients developed pneumonia in the perioperative period. During the follow-up period in the first year after surgery, no implant related adverse reaction was observed, including implant rupture, implant shift, rejection reaction and allergies. One patient had wound ulcer after chemotherapy. Three patients had tumor recurrence, with the recurrence rate of 25.0%. Two patients died of tumor recurrence, with a mortality rate of 16.7%. Conclusion    3DP titanium implant is a safe and effective material for chest wall reconstruction. 

OBJECTIVE: To summarize the experience of perioperative prevention during double-lung transplantation for elderly patients with coronavirus disease 2019 (COVID-19). METHODS: Clinical data of 2 elderly patients with COVID-19 who underwent double-lung transplantation in the First Affiliated Hospital of Zhejiang University School of Medicine in March 2020 were retrospectively reviewed. Perioperative protective measures were introduced in terms of medical staffing, respiratory tract, pressure injuries, air in operating room, instruments and equipment, pathological specimens, and information management. RESULTS: Two cases of double-lung transplantation were successfully completed, and the patients had no operation-related complications. Extracorporeal membrane oxygenator was successfully removed 2 to 4 days after surgery and the patients recovered well. There was no infection among medical staff. CONCLUSIONS Adequate preoperative preparation, complete patient transfer procedures, proper placement of instruments and equipment, strengthening of intraoperative care management, and attention to prevention of pressure injury complications can maximize the safety of COVID-19 patients and medical staff.
Aged ; Betacoronavirus ; COVID-19 ; Coronavirus Infections ; Humans ; Lung Transplantation/standards* ; Pandemics ; Perioperative Care/standards* ; Pneumonia, Viral ; Postoperative Complications/prevention & control* ; Retrospective Studies ; SARS-CoV-2 ; Transplant Recipients

Objective:To observe the presence or absence of necroptosis in PC12 cells after radiation injury, and to detect the expression of receptor-interacting protein 3(RIP3) and evaluate its regulatory effect on necroptosis.Methods:PC12 cells were treated with different doses of irradiation and their necroptosis was detected by lactate dehydrogenase (LDH) release at different time points. After pretreatment with necroptosis inhibitor Necrostatin-1(Nec-1), the changes of cell necroptosis were detected by LDH. The expression level of RIP3 after irradiation intervention was detected by Western blot (WB). After pretreatment with the RIP3-specific inhibitor GSK′872, the changes of cell necroptosis were detected by LDH. The best transfection sequence of RIP3 knockout was screened by WB. The cells were divided into the control group, irradiation group, solvent control group, no-load control group and pretreatment group. WB, immunofluorescence staining, MTT, LDH and Annex V-fluorescein Isothiocyanate/Propidium Iodide (AnnexV-FITC/PI) flow cytometry were used for detection and analysis.Results:After 4 Gy irradiation, the degree of cell necrosis was the highest after 3 hours of culture, and the expression level of RIP3 protein was up-regulated. The cell necrosis was decreased after Nec-1, GSK′872 and RIP3 gene knockdown pretreatment.Conclusions:The radiation injury of 4 Gy can induce the necroptosis of PC12 cells, and the most significant effect can be observed when cultured for 3 hours after irradiation. RIP3 is involved in the process of necroptosis of PC12 cells induced by radiation injury, and plays a pivotal positive regulatory role.