African swine fever(ASF)is an acute and highly pathogenic hemorrhagic disease of pigs,causing huge economic losses to pig industry.In order to quantitatively detect clinical samples of ASF and inactivated ASFV antigens,the IgG of ASF positive serum was used as capture anti-body and the HRP-labeled p72 monoclonal antibody was used as detecting antibody.The standard curve was drawn with the cell-cultured ASFV,and a sandwich ELISA detection of antigen was es-tablished.The specificity,sensitivity and stability of the method were evaluated.The effects of dif-ferent inactivation methods and adjuvant addition on antigen detection were further evaluated.The results showed that the minimum detection limits of the recombinant protein and the ASFV were 0.1 mg/L and 103.7 TCID50/mL,respectively.There was no cross-reaction with five common porcine pathogenic viruses,and the coefficient variations between batches was less than 10％.The total co-incidence rate with real-time fluorescence quantitative PCR was 92％(23/25).The sensitivity of antigen detection was significantly reduced when antigen was treated by BEI inactivation,and the detection results were severely interfered by aluminum adjuvant and nano-adjuvant.In summary,the sandwich ELISA antigen detection method established is specific,sensitive,and repeatable,with a good consistency to the qPCR method,which provides an effective clinical diagnostic meth-od for ASFV antigen.

In order to obtain a cell line which high expressed of rabies virus glycoprotein the produc-tion of rabies subunit vaccine,the RABV glycoprotein gene sequence was amplified by RT-PCR and cloned into the lentiviral expression vector.The recombinant plasmid pLV-RVG,envelope plasmid pMD2.0G and packaging plasmid psPAX2 were co-transfected into HEK-293T cells to harvest lentivirus and infect new HEK-293T cells.The 293T-RVG cell line was obtained by puro-mycin pressure screening.Indirect immunofluorescence assay(IFA)and Western blot showed that the 67 kD RABV-G protein was highly expressed in the cells.The neutralizing antibody titer reached 2.19 IU/mL within 14 days after immunization,which was higher than the internationally recognized protection threshold(0.5 IU/mL).This study showed that the cell line 293T-RVG with stable and high expression of rabies virus glycoprotein was successfully constructed.The cell line could protect mice against rabies virus challenge and could be used as a subunit vaccine with poten-tial for large-scale production and application.

Objective    To explore the potential role of tumor spread through air spaces (STAS) as a prognostic indicator of non-small cell lung cancer (NSCLC) through meta-analysis. Methods    PubMed, EMbase and Web of Science, from inception to February 2022 were searched by computer about the research of the 5-year overall survival (OS) and recurrence free survival (RFS) of NSCLC patients with or without STAS. The Newcastle-Ottawa scale (NOS) was used to evaluate the quality of each study. Results    Totally 13 published articles were included with 4 647 patients, and 1 424 (30.6%) patients had STAS. The NOS score of all studies≥6 points. The meta-analysis showed that compared with the NSCLC patients without STAS, those with STAS had a worse prognosis of 5-year RFS, and the combined HR was 1.89 (95%CI 1.61-2.23); they had a shorter 5-year OS, and the combined HR was 2.25 (95%CI 1.79-2.84). There was no statistical heterogeneity among studies. Conclusion    The presence of STAS may be a poor prognostic factor for patients with NSCLC, and enough attention should be paid. The STAS should be recorded in the pathological report to guide the comprehensive treatment and evaluate the prognosis of patients.

Acetylcholinesterase (AChE) is a key enzyme used to detect organophosphorus pesticide residues by the enzyme inhibition method. An accidental discovery of a mutant strain with AChE activity was made in our laboratory during the process of AChE expression by

Objective To compare GM260 portable blood glucose meter and AU5821 automatic biochemical analyzer in order to prove the accuracy of GM260 and its applicability for clinical use.Methods Totally 20 pieces of EDTA-K2 anticoagulative specimens and 23 GM260 meters were numbered,and each specimen underwent examinations by both GM260 and AU5821,then the bias between the two kinds of devices was calculated.Results The maximal bias between GM260 and AU5821 was 0.47 mmol/L and all the meters had the bias between-0.83 and 0.83 mmol/L in case of 5 specimens with the glucose concentration less than 4.2 mmol/L;the maximal bias between GM260 and AU5821 was 18.07％ and all the meters had the bias between-20％ and 20％ in case of 15 specimens with the glucose concentration not less than 4.2 mmol/L;the examination results by GM260 all accorded with industrial standard.The results by GM260 were lower than those by AU5821,and the maximal negative deviation was-13.43％.Conclusion Portable blood glucose meter can only be used for screening,and automatic biochemical analyzer is the preferred device for diabetes diagnosis.

Background and purpose:Visceral pleural invasion (VPI) and vessel invasion (VI) are poor prognostic factors in patients with non-small cell lung cancer (NSCLC). The primary initial recurrence site may be local recurrence in VPI and distant metastasis in VI. The purpose of this study was to validate the prognostic impact and effect of the initial recurrence site of VPI and VI on survival outcomes for NSCLC.Methods:Two hundred and ninety patients who were diagnosed as having NSCLC and underwent lobectomy between Jan. 2007 and Dec. 2013 were retrospectively analyzed. VPI was identiifed in 51 patients as VPI group, the other 239 patients without VPI as non-VPI group. VI was identiifed in 29 patients as VI group, the other 261 patients without VI as non-VI group. Clinical characteristics, overall survival (OS), disease-free survival (DFS) were compared.Results:There were statistically signiifcant differences between VPI group and non-VPI group in tumor size, lymph node metastasis, TNM stage and initial recurrence site (P<0.05). Furthermore, there were statistically signiifcant differences between VI group and non-VI group in lymph node metastasis and TNM stage (P<0.05). The 1-, 3- and 5-year OS rates in VPI group (88.2%, 56.7% and 52.7%) were lower than those in non-VPI group (95.8%, 83.7% and 74.0%,P<0.001). The 1-, 3- and 5-year OS rates in VI group (79.3%, 56.8% and 48.7%) were lower than those in non-VI group (96.1%, 81.3% and
72.3%,P=0.001). Cox regression showed TNM stage was a significant prognostic factor for DFS, whereas lymph node metastasis and VPI were signiifcant prognostic factors in patients with NSCLC.Conclusion:The primary initial recurrence site in VPI patients is local recurrence. Patients with VPI or VI may need more postoperative therapy because of their poor prognosis.

OBJECTIVETo investigate the expression of IFITM3 in colorectal carcinoma and its clinical significance.

METHODS213 patients with colon ademocarcinoma and 214 patients with colon adenoma treated by surgery in our hospital from March 2008 to June 2010 were included in this study. The levels of IFITM3 in normal colon nucosa, adenoma, and adenocarcinoma tissues were detected by real-time PCR and immunochemistry, and its relationship with metastasis and prognosis in 213 colorectal cancer patients was analyzed.

RESULTSThe IFITM3 mRNA level in metastatic tumor group was 18.37 ± 0.61, significantly higher than that in the normal 4.49 ± 0.69 and non-metastases groups (7.32 ± 0.76; F = 460.380, P < 0.001). The positive rate of IFITM3 protein expression in metastatic tumor group (69.0%) was significantly higher than that in the normal (3.9%), non-metastasies groups (19.0%) and adenoma groups (11.3%). Our clinical analysis confirmed that the IFITM3 expression was associated with peritumoral invasion, hepatic metastases, metastases of para-colonic lymph nodes, mesocolonic lymph nodes and mesenteric root lymph nodes, omental metastasis and AJCC classification (P < 0.05). Furthermore, the survival curve analysis showed that patients with lower IFITM3 level expression had a higher 5-year survival rate (88.8%) than that in the patients with higher expression (40.2%, P < 0.001).

CONCLUSIONSIFITM3 expression has a positive correlation with metastasis and prognosis in patients with colorectal carcinoma.

Adenocarcinoma ; diagnosis ; metabolism ; Adenoma ; Colorectal Neoplasms ; diagnosis ; metabolism ; Humans ; Liver Neoplasms ; metabolism ; Membrane Proteins ; genetics ; metabolism ; Neoplasms ; Peritoneal Neoplasms ; Prognosis ; RNA, Messenger ; RNA-Binding Proteins ; genetics ; metabolism ; Real-Time Polymerase Chain Reaction ; Survival Analysis ; Survival Rate

OBJECTIVETo inhibit the proliferation and metastasis of colon cancer cells by increasing the expression level of B-cell translocation gene-2 (BTG2).

METHODSWestern blot assay was used to detect the expression level of BTG2 protein in the normal intestinal epithelial HIEC cells and three colon cancer cell lines SW620, HT-29 and LS174T. The expression of BTG2 protein in normal colonic epithelial tissue, colon adenoma and colon cancer tissue was detected by immunohistochemistry. The plasmid with BTG2 gene full-length sequence was transfected into colon cancer SW620 cells, and the expression of BTG2 protein was detected by Western blot. The cell growth curve was drawn by MTT test. The Ki-67-positive rate was calculated using immunofluorescence staining. The cell migration of colon cancer cells was detected by scratch test and Transwell double chamber culture system, and the pseudopodia growth of tumor cells was detected by Matrigel 3D culture system.

RESULTSWestern blot results showed that BTG2 relative expression levels were 0.83 ± 0.12, 0.18 ± 0.04, 0.20 ± 0.05 and 0.36 ± 0.07 in normal human intestinal epithelial cells HIEC, and human colon cancer cell line SW620, HT-29 and LS174T, respectively. The results of immunohistochemistry showed that the positive expression of BTG2 protein in normal colorectal tissue, colorectal adenoma and colorectal carcinoma tissues were 82.5% (33/40), 77.5%(31/40) and 17.5% (7/40), respectively, with a significant difference between two groups (P < 0.05). Immunofluorescence results showed that the positive rate of Ki-67 in the control group, empty vector group and BTG2 transfection group was (76.2 ± 8.0)%, (81.4 ± 9.7)% and (50.1 ± 7.1)%, respectively, showing a significant difference between two groups (P < 0.05). The scratch test results showed that in the control group, empty vector group and BTG2 transfection group, the distance of SW620 cells between two sides was (79.27 ± 11.24) µm, (80.65 ± 12.17) µm and (124.77 ± 19.63) µm, respectively, with a significant difference between two groups (P < 0.05). Transwell results showed that in the control group, empty plasmid group and BTG2 transfection group, the SW620 cell migration rate was (78.5 ± 13.1)%, (73.2 ± 12.9)% and (47.4 ± 9.1)%, respectively, showing a significant difference between two groups (P < 0.05). The number of neurospheres of BTG2 transfection group was decreased SW620, which had poor ductility.

CONCLUSIONSBTG2 gene is involved in colon cancer cell proliferation and metastasis, and effectively restores the function of BTG2 protein. Therefore, it may be expected to become a new option in gene therapy for colon cancer.

B-Lymphocytes ; physiology ; Cell Cycle ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; genetics ; Colonic Neoplasms ; Genetic Vectors ; Humans ; Immediate-Early Proteins ; genetics ; Immunohistochemistry ; Plasmids ; Transfection ; Tumor Suppressor Proteins ; genetics

[ABSTRACT]OBJECTIVETo investigate the diagnosis, treatment method, key points of operation, and postoperative complications of high-risk esophageal foreign body.METHODSA retrospective analysis of 41 cases of high-risk esophageal foreign body from January 1996 to December 2014. After adequate preparation, the foreign body was removed via esophageal endoscope under general anesthesia.RESULTSThe foreign bodies in 41 patients were removed via esophageal endoscope once or twice. Two cases suffered postoperative subcutaneous emphysema, that may be a result of a small perforation in esophagus. Emphysema was disappeared by fast, rehydration and anti-infection for 6 to 8 days, and other serious complications did not occur.CONCLUSIONMost high-risk esophageal foreign bodies can be removed through rigid esophagoscopy. Some of the foreign bodies of the patients were difficult to remove, some patients were presented with mediastinal emphysema and pneumothorax due to esophageal perforation, and some foreign body stuck in oesophagus so long to cause esophageal mucosa ulcer. In these conditions, foreign bodies should be removed by lateral neck incision or thoracotomy.

BACKGROUND:Previously deep burn wound or skin defects are generaly repaired with skin grafting or flap of skin grafting. Obvious scar hyperplasia usualy appears after operation, which requires multiple surgeries. Meanwhile, patients have to suffer from great pain and bear high cost. OBJECTIVE: To observe the clinical effects on deep wounds by continuous traction of self-designed skin-stretching device (patent No. ZL 2012 2 0022443.7). METHODS: Thirty patients with deep burn wound, skin defect or funicular scar were enroled, including 22 males and 8 females, aged 18-49 years, and randomly divided into two groups. Skin-stretching device was adopted for skin traction treatment. Twenty cases underwent skin traction from 1 kg puling force to 5 kg, with an increase of 1 kg per 2 days, 6 hours a day for 10 days. Blood flow at the beginning, 1, 5, 10, 15, 20, 30, 60 minutes of the skin traction, and the changes of wound edge skin as wel as histological changes of the skin were observed. Of the remaining 10 cases, 2, 6, and 2 cases underwent skin traction of 2, 4, 7 kg, respectively. Blood flow and skin changes were also observed to find out the most suitable and safe force. RESULTS AND CONLUSION:Al the 30 cases achieved primary healing without necrosis of skin, infection or peripheral circulatory disorders, and the appearance and function recovered wel. The healing time was 8-24 days. The skin-stretching device was most safe under 4 kg puling force, by which, there was neither blood circulation obstacle nor tear of skin. After traction, the skin blood flow and the number of cels increased, especialy the epithelial basal cels. The colagen fibers became thicker and denser, and the elastic fibers regenerated significantly; the fibroblasts and capillary density increased. It has been proved that we can better close the wound and reduce scar formation effectively with the self-designed skin-stretching device for skin traction.