Objective To analyze the confirmation results of initial screening positive for human immunodeficiency virus (HIV) antibodies and to investigate the rationality of the gray zone for the signal-to-cutoff (S/CO) ratio values obtained by chemiluminescence assay. Methods A total of 184 specimens with positive HIV antibody screening results were collected. Confirmation was conducted using the Western blot (WB) method and viral nucleic acid testing, and the confirmation outcomes and band distribution were observed. The receiver operating characteristic (ROC) curve was employed to predict the optimal cutoff value, and the rationality of S/CO values in establishing a gray zone for HIV antibody was analyzed. Results Among the 184 HIV antibody screening positive samples, 153(83.15%) were initially screened by chemiluminescence, and 31 (16.85%) were screened by colloidal gold method. Male cases (57.07%) and those aged 40 years or older (75.00%) had higher proportion of initial screening positivity than females and other age groups. Supplemental WB testing revealed that 101 (54.89%) cases were confirmed as HIV-1 antibody positive, 56(30.43%) were negative, and 27(14.67%) were indeterminate. Of the 27 indeterminate cases, one was confirmed positive by NAT (initial S/CO value by chemiluminescence5.60), while the remaining 26 were negative. The positive agreement rates between chemiluminescence, colloidal gold and supplemental WB testing were 65.36% and 3.23%, respectively, with statistically significant difference (χ²=40.191, P < 0.05). Among 27 cases with indeterminate HIV-1 antibody results, upon further nucleic acid testing for viral load, one case exhibited WB banding pattern of P24+gp160, and the nucleic acid load confirmed this case as positive. In the distribution of band patterns among indeterminate samples, P24 had the highest occurrence (74.07%), followed by P66 (11.11%), gp160 (7.41%) as well as P17 and gp41 (both 3.70%). The ROC curve predicted the optimal S/CO cutoff value to be 5.15, with an area under the curve (AUC) of 0.904, sensitivity and specificity of 83.30% and 76.40%, respectively, and 95%CI of 0.861 to 0.948. There was a good correlation between the WB band patterns and confirmation results for initial screening positive cases (P < 0.05), with the highest correlation found for the P24 band (r=0.910). Conclusion There is a certain rate of false positives among HIV antibody initial screening positive cases, and effective screening can be achieved through WB testing. Additionally, based on the current testing platform, the gray zone for HIV antibody S/CO values in the chemiluminescence assay is set at 1.00 to 5.15, which exhibits high sensitivity.

Objective:To investigate the application value of self-pulling and latter transection (SPLT) technique in double anti-reflux double-tract reconstruction of totally laparoscopic proximal gastrectomy.Methods:The retrospective cohort study was conducted. The clinicopatholo-gical data of 103 patients with Siewert type Ⅱ adenocarcinoma of esophagogastric junction in clinical stage Ⅰ-Ⅱ who were admitted to Shanxi Cancer Hospital from January 2018 to January 2020 were collected. There were 65 males and 38 females, aged from 45 to 79 years, with a median age of 59 years. Of 103 patients, 49 cases undergoing totally laparoscopic proximal gastrectomy with double-tract reconstruction of SPLT were assigned into the SPLT group, 54 cases undergoing totally laparoscopic proximal gastrectomy with conventional double-tract reconstruction were assigned into the traditional group. Observation indicators: (1) intraoperative situations; (2) postoperative situations; (3) follow-up. Follow-up was conducted by outpatient examination and telephone inter-view to detect postoperative reflux esophagitis of patients up to December 2021. Measurement data with normal distribution were represented as Mean± SD, and the t test was used for comparison between groups. Measurement data with skewed distribution were represented as M(range) or M( Q1, Q3), and the Wilcoxon test was used for comparison between groups. Count data were described as absolute numbers or percentages, and comparison between groups was performed using the chi-square test. Comparison of ordinal data was analyzed using the non-parameter rank sum test. Results:(1) Intraoperative situations: the operation time, digestive tract reconstruction time, volume of intraoperative blood loss, the number of inferior mediastinal lymph nodes dissected, cases with auxiliary incisions for the SPLT group were (261±48)minutes, (26±4)minutes, (114±42)mL, 8.0(6.5,9.5), 1, respectively. The above indicators were (244±42)minutes, (30±6)minutes, (118±46)mL, 5.5(4.0,8.0), 9 for the traditional group, respectively. There were significant differences in the digestive tract reconstruction time, the number of inferior mediastinal lymph nodes dissected and cases with auxiliary incisions between the two groups ( t=-3.34, Z=-4.05, χ2=4.72, P<0.05). There was no significant difference in the operation time or volume of intraoperative blood loss between the two groups ( t=1.87, -0.47, P>0.05). (2) Postoperative situations: duration of postopera-tive hospital stay and cases with postoperative complications were (11.5±2.7)days and 4 for the SPLT group, versus (12.5±4.3)days and 9 for the traditional group, showing no significant difference between the two groups ( t=-1.47, χ2=1.68, P>0.05). There were 13 of 103 patients with postopera-tive complications, including 5 cases of left pleural effusion, 4 cases of anastomotic leakage, 2 cases of mild pneumonia, 1 case of incision infection, 1 case of chylous leakage. Four patients had anasto-motic leakage at the esophagojejunostomy, the abdominal esophagus of whom was invaded by more than 1 cm. During the operation, mediastinal drainage tubes were placed through the abdominal wall. The 4 patients were cured after enteral and parenteral nutrition support and adequate drainage, and the remaining patients with complications were cured after symptomatic treatment. (3) Follow-up: of 49 patients in the SPLT group, 43 cases were followed up for (18±4)months. During the follow-up, 1 case showed reflux esophagitis by gastroscopy, with the incidence of 2.33%(1/43). Of 54 patients in the traditional group, 53 cases were followed up for (17±4)months. During the follow-up, 4 cases showed reflux esophagitis by gastroscopy, with the incidence of 7.55%(4/53). There was no significant difference in the incidence of reflux esophagitis between the two groups ( χ2=0.47, P>0.05). Conclusions:SPLT technology is feasible for double anti-reflux double-tract reconstruction of proximal gastrectomy. Compared with traditional double-tract reconstruction of totally laparos-copic proximal gastrectomy, SPLT technology can reduce the auxiliary incisions, increase the number of lower mediastinal lymph nodes dissected, and shorten the digestive tract reconstruction time.

Objective:To investigate effect of reperfusion injury following different ischemic duration on skeletal muscle in rats.Methods:A model of ischemia/reperfusion injury (IRI) was established by unilateral clamping femoral artery and additional application of tourniquet in skeletal muscle of hind limbs in 35 male Wsitar rats. According to different ischemia time, the animals were assigned to 2-hour ischemia and 24-hour reperfusion (I2R24 group), 2.5-hour ischemia and 24-hour reperfusion (I2.5R24 group), 3-hour ischemia and 24-hour reperfusion (I3R24 group), 4-hour ischemia and 24-hour reperfusion (I4R24 group) and sham group, with 7 rats per group. At the end of reperfusion, gastrocnemious tissues and plasma samples were collected and analyzed. The ratio of wet ∶ dry weight (W/D) was used to measure muscle edema. The assay of 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) was conducted to evaluate muscle viability. HE staining was executed to observe histopathological changes. Immunofluorescence staining was performed to assess the levels of C1q, C3b/c, tissue factor (TF), fibrinogen (FN), bradykinin receptor 1 (BR1), BR2, vascular cell adhesion molecule-1 (VCAM-1), E-selectin, fibrinogen-like protein-2 (FGL-2) and myeloperoxidase (MPO) in muscle tissues. ELISA method was used to determine the concentrations of interferonγ (IFN-γ), interleukin7 (IL-7), IL-18, macrophage inflammatory1α (MIP-1α) and monocyte chemotactic protein 1 (MCP-1) in plasma.Results:With prolongation of ischemia time and subsequent reperfusion, tissue edema became severe gradually. The ratio of W/D was 5.3±0.2, 6.1±0.3, 6.9±0.2, 7.6±0.3 in I2R24, I2.5R24, I3R24 and I4R24 groups, higher than that in sham group (4.5±0.1) (all P<0.01). Muscle viability got decreased gradually. Muscle viability was (62.4±3.5)%, (45.3±3.3)%, (35.4±3.4)%, (27.1±5.9)% in I2R24, I2.5R24, I3R24 and I4R24 groups, lower than that in sham group［(93.8±7.2)%］(all P<0.01). Histopathological changes became aggravated gradually. The most severe group was I4R24 group, with the most severe myocyte injury, interstitial edema and extensive inflammatory infiltration, followed by I3R24, I2.5R24 and I2R24 groups in order. There was normal structure integrity and neatly arranged myocyte in sham group. Meanwhile, levels of C1q, C3b, FN, BR1, VCAM-1, E-selectin and FGL-2 got increased gradually. The highest levels for these factors were seen in I4R24 group, followed by I3R24 group, I2.5R24 group, I2R24 group and sham group in order. The rough ratio of the number of positive MPO cells/total cell number under high lens (×200) were increased gradually, with the highest level in I4R24 group, followed by I3R24 group, I2.5R24 group, I2R24 group and sham group in order. However, expression of TF and BR2 were not altered significantly among the groups. Plasma levels of INF-γ, IL-7, IL-18, MIP-1α and MCP-1 elevated gradually with prolongation of ischemia time (all P<0.01). The sequence was the sham group, I2R24 group, I2.5R24 group, I3R24 group and I4R24 groups for levels of these factors from low to high (all P<0.01). Conclusion:Reperfusion after prolongation of ischemia duration can increase the activation of complement, coagulation, kinin and endothelial cells as well as the release of inflammatory factors, and thus aggravate the degree of skeletal muscle tissue injury.

Objective:To evaluate the safety and efficiency of a modified double-tract reconstruction procedure─proximal gastrectomy with piggyback interposed jejunal single-channel reconstruction (PJIR-STR) for early SiewertⅡ adenocarcinoma of esophagogastric junction (AEG).Method:Data of 8 SiewertⅡ AEG patients at Shanxi Tumor Hospital and undergoing PJIR-STR from May 2018 to Oct 2019 were retrospectively analyzed. The gastroesophageal reflux disease questionnaire (GerdQ) was used to score the patients at 3, 6, 12, and 18 months after surgery. The severity of postoperative reflux esophagitis was assessed by gastroscopy at 3, 6 months after surgery, using the Los Angeles Classification criteria.Result:All patients recovered well after surgery without serious complications. No obvious gastroesophageal reflux was observed in all patients at different periods (All of the GerdQ scores were <8 points.) The results of gastroscopy showed that 1 patient was diagnosed as grade B reflux esophagitis at 3, 6 months after surgery, which was responsive to conservative treatment, and the other 7 patients had no grade B or above reflux esophagitis.Conclusion:PJIR-STR is a feasible, safe reconstruction with excellent efficiency of dual anti-reflux for the SiewertⅡ AEG.

As a novel post-translational modification(PTM),lysine 2-hydroxyisobutyrylation(Khib)is considered to regulate gene transcriptional activities in eukaryotic cells;however,the functions of Khib-modified proteins in plants remain unknown.Here,we report that Khib is an evolutionarily-conserved PTM in wheat and its progenitors.A total of 3348 Khib sites on 1074 proteins are iden-tified in common wheat(Triticum aestivum L.)by using affinity purification and mass spectroscopy of 2-hydroxyisobutyrylome.Bioinformatic data indicate that Khib-modified proteins participate in a wide variety of biological and metabolic pathways.Immunoprecipitation confirms that Khib-modified proteins are present endogenously.A comparison of Khib and other main PTMs shows that Khib-modified proteins are simultaneously modified by multiple PTMs.Using mutagenesis experiments and co-immunoprecipitation assays,we demonstrate that Khib on K206 of phospho-glycerate kinase(PGK)is a key regulatory modification for its enzymatic activity,and mutation on K206 affects the interactions of PGK with its substrates.Furthermore,Khib modification of low-molecular-weight proteins is a response to the deacetylase inhibitors nicotinamide and tricho-statin.This study provides evidence to promote our current understanding of Khib in wheat plants,including the cooperation between Khib and its metabolic regulation.

There are still many controversies in the surgical treatment of esophagogastric junction tumors in terms of surgical approach, cleaning range, and resection scope. The reason is the confusion about the scope of the esophagogastric junction. The previous domestic and foreign anatomy descriptions of this part are not enough to solve the current problems. Based on a large number of basic anatomy and clinical operations, this article proposes that the esophagogastric junction may be wrapped by a complete and separate esophagogastric junction membrane with independent anatomy other than infracardiac bursa. The structure of the transitional tissue, mainly from the distribution of submucosal veins, explained the relationship and significance of tissue transitional changes and clinical operations, and made a reasonable analysis of the current controversy based on the anatomical characteristics, which is worthy of further investigation.

There are still many controversies in the surgical treatment of esophagogastric junction tumors in terms of surgical approach, cleaning range, and resection scope. The reason is the confusion about the scope of the esophagogastric junction. The previous domestic and foreign anatomy descriptions of this part are not enough to solve the current problems. Based on a large number of basic anatomy and clinical operations, this article proposes that the esophagogastric junction may be wrapped by a complete and separate esophagogastric junction membrane with independent anatomy other than infracardiac bursa. The structure of the transitional tissue, mainly from the distribution of submucosal veins, explained the relationship and significance of tissue transitional changes and clinical operations, and made a reasonable analysis of the current controversy based on the anatomical characteristics, which is worthy of further investigation.

Objective:To detect and verifica the gene profile difference of microRNA-146a (miR-146a) and its role in the pro-liferation of vascular smooth muscle cells (VSMCs) by gene chip technology. Methods: Artificially synthesized miR-146a mimics(50 nmol/L) ,miR-146 inhibitor ( 50 nmol/L ) , scramble ( 50 nmol/L ) and PBS were transfected into cultured primary rat VSMCs in vitro. After transfection,Real time PCR was used to measure the levels of miR-146a and the cell counting kit 8(CCK8) was employed to investigate the proliferation of VSMCs. The VSMCs interfered by miR-146a inhibitor or miR-146a control were examined by gene chips and the profile of gene were analyzed by bioinformatics technology to detect the different genes and signal transduction pathway. The changes in mRNAs and proteins were accessed separately by Real time PCR and Western blot. Results: Compared with sham and control VSMCs,miR-146a expression level was significantly decreased in treatment with miR-146a inhibitor(P<0. 01),as well as optical density(OD) was also shown remarkably down regulated simultaneously(P<0. 05). The investigation of gene profile revealed that the p53 signal pathway was up-regulated in VSMCs interfered by miR-146a. The mRNA and protein expression levels of p53, caspase3 and PTEN in p53 signal transduction pathway didn′t show significant differences(P>0. 05),however,the mRNA and protein expression levels of cyclin D1 significantly increased in treatment with miR-146a mimics VSMCs group and decreased in miR-146a inhibitor VSMCs group ( compared with sham VSMCs group, both P<0. 05 ) . Conclusion: Our data indicated that miR-146a may promote the proliferation of rat VSMCs by up-regulating cyclin D1 expression.

AIM:To investigate the proliferation property of stable chemerin gene knockdown vascular smooth muscle cells ( VSMCs) and to explore its mechanism .METHODS:The normal VSMCs , chemerin gene interfering control VSMCs and stable chemerin gene knockdown VSMCs were divided into normal group , PDGF group, control group and knockdown group .The VSMCs in PDGF group were given platelet-derived growth factor-BB ( PDGF-BB) to initiate proli-feration.The cell counting and BrdU assay were employed to investigate the proliferation property of VSMCs .The mitogen-activated protein kinase ( MAPK) signal pathway was determined by Western blot .RESULTS:The cell number and BrdU A value in PDGF-BB-treated VSMCs significantly increased as compared with the normal VSMCs ( P<0.05 ) .Compared with the normal VSMCs , the chemerin knockdown VSMCs showed obviously decreased cell number and BrdU A value ( P<0.05).Simultaneously, no significant difference in the proliferation of VSMCs between the normal VSMCs and the control VSMCs was observed.No significant difference of the protein levels of p-ERK1/2, ERK1/2, p-p38 MAPK and p38 MAPK among 4 kinds of VSMCs was found .The protein level of p-JNK in PDGF-BB-treated VSMCs was up-regulated, while it was down-regulated in chemerin knockdown VSMCs compared with the normal VSMCs .CONCLUSION: Chemerin pro-motes the proliferation of mouse vascular smooth muscle cells by up-regulating p-JNK production .

AIM: To observe the effects of H_2O_2 on apoptosis of skeletal muscle satellite cells (SMSC)and mitochondrial membrane potential (MMP), and protective effect of erythropoietin(EPO). METHODS: SMSC in vitro were divided into three groups: H_2O_2 group, H_2O_2+EPO group and control. Apoptosis rate and the means were obverted by monofluorescence flow cytometry. The morphological change of apoptosis cells were observed under fluorescence microscopy after Hoechst 33258 staining. RESULTS: The cells in H_2O_2 group show the highest apoptosis rate (22.13±1.79)%. In H_2O_2+EPO group, apoptosis rate were (16.47±2.53)%, (4.97±0.55)% and (2.93±0.47)% according to the EPO treated levels (10, 20 or 40 kU/L), respectively. MMP level in H_2O_2 group was the lowest 9.70±0.09. MMP levels in H_2O_2+EPO group were 12.67±0.32, 27.90±0.66, 44.53±0.93, respectively according to the EPO treated levels (10, 20 or 40 kU/L). In control group, apoptosis rate was 1.93±0.57 and MMP was 51.37±0.64. In H_2O_2 group and H_2O_2+ low dosage EPO group, Hoechst 33258 staining showed obvious apoptosis. CONCLUSION: EPO inhibits the apoptosis induced by H_2O_2 and stabilizes the MMP, which is related to the dosage of EPO.