[Safety of double and a half layered esophagojejunal anastomosis in radical gastrectomy: A prospective, multi-center, single arm trial].

Objective: To evaluate the safety of double and a half layered esophagojejunal anastomosis in radical gastrectomy. Methods: This prospective, multi-center, single-arm study was initiated by the Affiliated Cancer Hospital of Zhengzhou University in June 2021 (CRAFT Study, NCT05282563). Participating institutions included Nanyang Central Hospital, Zhumadian Central Hospital, Luoyang Central Hospital, First Affiliated Hospital of Henan Polytechnic University, First Affiliated Hospital of Henan University, Luohe Central Hospital, the People's Hospital of Hebi, First People's Hospital of Shangqiu, Anyang Tumor Hospital, First People's Hospital of Pingdingshan, and Zhengzhou Central Hospital Affiliated to Zhengzhou University. Inclusion criteria were as follows: (1) gastric adenocarcinoma confirmed by preoperative gastroscopy;(2) preoperative imaging assessment indicated that R0 resection was feasible; (3) preoperative assessment showed no contraindications to surgery;(4) esophagojejunostomy planned during the procedure; (5) patients volunteered to participate in this study and gave their written informed consent; (6) ECOG score 0-1; and (7) ASA score I-III. Exclusion criteria were as follows: (1) history of upper abdominal surgery (except laparoscopic cholecystectomy);(2) history of gastric surgery (except endoscopic submucosal dissection and endoscopic mucosal resection); (3) pregnancy or lactation;(4) emergency surgery for gastric cancer-related complications (perforation, hemorrhage, obstruction); (5) other malignant tumors within 5 years or coexisting malignant tumors;(6) arterial embolism within 6 months, such as angina pectoris, myocardial infarction, and cerebrovascular accident; and (7) comorbidities or mental health abnormalities that could affect patients' participation in the study. Patients were eliminated from the study if: (1) radical gastrectomy could not be completed; (2) end-to-side esophagojejunal anastomosis was not performed during the procedure; or (3) esophagojejunal anastomosis reinforcement was not possible. Double and a half layered esophagojejunal anastomosis was performed as follows: (1) Open surgery: the full thickness of the anastomosis is continuously sutured, followed by embedding the seromuscular layer with barbed or 3-0 absorbable sutures. The anastomosis is sutured with an average of six to eight stitches. (2) Laparoscopic surgery: the anastomosis is strengthened by counterclockwise full-layer sutures. Once the anastomosis has been sutured to the right posterior aspect of the anastomosis, the jejunum stump is pulled to the right and the anastomosis turned over to continue to complete reinforcement of the posterior wall. The suture interval is approximately 5 mm. After completing the full-thickness suture, the anastomosis is embedded in the seromuscular layer. Relevant data of patients who had undergone radical gastrectomy in the above 12 centers from June 2021 were collected and analyzed. The primary outcome was safety (e.g., postoperative complications, and treatment). Other studied variables included details of surgery (e.g., surgery time, intraoperative bleeding), postoperative recovery (postoperative time to passing flatus and oral intake, length of hospital stay), and follow-up conditions (quality of life as assessed by Visick scores). Result: [1] From June 2021 to September 2022,457 patients were enrolled, including 355 men and 102 women of median age 60.8±10.1 years and BMI 23.7±3.2 kg/m2. The tumors were located in the upper stomach in 294 patients, mid stomach in 139; and lower stomach in 24. The surgical procedures comprised 48 proximal gastrectomies and 409 total gastrectomies. Neoadjuvant chemotherapy was administered to 85 patients. Other organs were resected in 85 patients. The maximum tumor diameter was 4.3±2.2 cm, number of excised lymph nodes 28.3±15.2, and number of positive lymph nodes five (range one to four. As to pathological stage,83 patients had Stage I disease, 128 Stage II, 237 Stage III, and nine Stage IV. [2] The studied surgery-related variables were as follows: The operation was successfully completed in all patients, 352 via a transabdominal approach, 25 via a transhiatus approach, and 80 via a transthoracoabdominal approach. The whole procedure was performed laparoscopically in 53 patients (11.6%), 189 (41.4%) underwent laparoscopic-assisted surgery, and 215 (47.0%) underwent open surgery. The median intraoperative blood loss was 200 (range, 10-1 350) mL, and the operating time 215.6±66.7 minutes. The anastomotic reinforcement time was 2 (7.3±3.9) minutes for laparoscopic-assisted surgery, 17.6±1.7 minutes for total laparoscopy, and 6.0±1.2 minutes for open surgery. [3] The studied postoperative variables were as follows: The median time to postoperative passage of flatus was 3.1±1.1 days and the postoperative gastrointestinal angiography time 6 (range, 4-13) days. The median time to postoperative oral intake was 7 (range, 2-14) days, and the postoperative hospitalization time 15.8±6.7 days. [4] The safety-related variables were as follows: In total, there were 184 (40.3%) postoperative complications. These comprised esophagojejunal anastomosis complications in 10 patients (2.2%), four (0.9%) being anastomotic leakage (including two cases of subclinical leakage and two of clinical leakage; all resolved with conservative treatment); and six patients (1.3%) with anastomotic stenosis (two who underwent endoscopic balloon dilation 21 and 46 days after surgery, the others improved after a change in diet). There was no anastomotic bleeding. Non-anastomotic complications occurred in 174 patients (38.1%). All patients attended for follow-up at least once, the median follow-up time being 10 (3-18) months. Visick grades were as follows: Class I, 89.1% (407/457); Class II, 7.9% (36/457); Class III, 2.6% (12/457); and Class IV 0.4% (2/457). Conclusion: Double and a half layered esophagojejunal anastomosis in radical gastrectomy is safe and feasible.

[Transcriptomic analysis of the ΔPaLoc mutant of Clostridioides difficile and verification of its toxicity].

Objective: Comparative analyses of wild-type Clostridioides difficile 630 (Cd630) strain and pathogenicity locus (PaLoc) knockout mutant (ΔPaLoc) by using RNA-seq technology. Analysis of differential expression of Cd630 wild-type strain and ΔPaLoc mutant strain and measurement of its cellular virulence changes. Lay the foundation for the construction of an toxin-attenuated vaccine strain against Clostridioides difficile. Methods: Analysis of Cd630 and ΔPaLoc mutant strains using high-throughput sequencing (RNA-seq). Clustering differentially expressed genes and screening differentially expressed genes by DESeq software. Further analysis of differential genes using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment. Finally, cytotoxicity assays of ΔPaLoc and Cd630 strains were performed in the African monkey kidney epithelial cell (Vero) and the human colonic cell (Caco-2) lines. Results: The transcriptome data showed that the ΔPaLoc mutant toxin genes tcdA and tcdB were not transcribed. Compared to the wild-type strain, CD630_36010, CD630_020910,CD630_02080 and cel genes upregulated 17.92,11.40,8.93 and 7.55 fold, respectively. Whereas the hom2 (high serine dehydrogenase), the CD630_15810 (spore-forming protein), CD630_23230 (zinc-binding dehydrogenase) and CD630_23240 (galactitol 1-phosphate 5-dehydrogenase) genes were down-regulated by 0.06, 0.075, 0.133 and 0.183 fold, respectively. The GO and KEGG enrichment analyses showed that the differentially transcribed genes in ΔPaLoc were enriched in the density-sensing system, ABC transport system, two-component system, phosphotransferase (PTS) system, and sugar metabolism pathway, as well as vancomycin resistance-related pathways. Cytotoxicity assays showed that the ΔPaLoc mutant strain lost its virulence to Vero and Caco-2 cells compared to the wild-type Cd630 strain. Conclusion: Transcriptional sequencing analysis of the Cd630 and ΔPaLoc mutant strains showed that the toxin genes were not transcribed. Those other differential genes could provide a reference for further studies on the physiological and biochemical properties of the ΔPaLoc mutant strain. Cytotoxicity assays confirmed that the ΔPaLoc mutant lost virulence to Vero and Caco-2 cells, thus laying the foundation for constructing an toxin-attenuated vaccine strain against C. difficile.

[Long term prognosis of fulminant myocarditis and predictors related to impaired cardiac function post discharge].

Objective: To compare the long-term prognosis of fulminant myocarditis (FM) and non-fulminant myocarditis (NFM) patients who survived and discharged from hospital, and to explore the factors associated with the long-term prognosis and impaired cardiac function. Methods: This study was a retrospective study. Consecutive patients with acute myocarditis hospitalized in Tongji Hospital from January 2017 to December 2020 were enrolled and divided into FM group and NFM group according to the type of myocarditis. Then, patients in the FM group were further divided into normal cardiac function group and impaired cardiac function group according the left ventricular ejection fraction (LVEF). All patients with acute myocarditis were treated with antiviral, immunomodulatory, immunosuppressive medications and symptomatic and supportive treatment, while FM patients were treated with comprehensive treatment plan. Clinical data at admission of enrolled patients were collected through the electronic medical record system. Patients were clinically followed-up at 1, 3, 6 and 12 months, then once a year after discharge by clinical visit. The primary endpoints included major cardiovascular events, impaired cardiac function was defined by LVEF<55%. Kaplan-Meier survival curve was used to analyze the occurrence of LVEF<55% and left ventricular enlargement during the follow-up of patients in FM group and NFM group, and Log-rank test was used for comparison between groups. Cox regression model was used to analyze the risk factors of impaired cardiac function in patients with FM during follow-up. Results: A total of 125 patients with acute myocarditis were enrolled (66 in FM group and 59 in NFM group). Compared with NFM group, the proportion of FM patients with the lowest LVEF<55% during hospitalization was higher (P<0.01), and the recovery time of normal LVEF during hospitalization was longer (P<0.01). The proportion of LVEF<55% at discharge was similar between the two groups (P=0.071). During the follow-up of 12 (6, 24) months, 1 patient (1.5%) died due to cardiac reasons in FM group after discharge, 16 patients (24.2%) had sustained LVEF<55% after discharge, and 8 patients (12.1%) had left ventricular enlargement. In NFM group, 3 patients (5.1%) had sustained LVEF<55%, and 1 patient (1.7%) had left ventricular enlargement. Kaplan-Meier survival curve analysis showed that the incidence of sustained LVEF<55% in FM group was higher than that in NFM group (P=0.003), and the incidence of left ventricular enlargement was also higher than that in NFM group (P=0.024). Subgroup analysis of patients in the FM group showed that, compared with the normal cardiac function group, the time from onset to admission was shorter (P=0.011), the proportion of LVEF<55% at discharge was higher (P=0.039), the proportion of coronary angiography was higher (P=0.014), and the LVEF recovery time during hospitalization was longer (P=0.036) in FM patients with impaired cardiac function. Multivariate Cox regression analysis showed that longer LVEF recovery time during hospitalization was an independent risk factor for cardiac function impairment after discharge of FM patients (HR=1.199, 95%CI 1.023-1.406, P=0.025). Conclusions: The incidence of reduced LVEF is significantly higher in FM patients than that in NFM patients. Longer LVEF recovery time during hospitalization is an independent risk factor for cardiac function impairment in FM patients after discharge.

CT and MRI features in 22 cases of primary malignant thymic germ cell tumours.

AIM: To analyse the computed tomography (CT) and magnetic resonance imaging (MRI) features of malignant thymic germ cell tumours (GCTs), in order to improve the accuracy of diagnosis of these tumours. MATERIALS AND METHODS: Twenty-two patients (20 men, two women; age, 28 ± 8.64 years) with malignant thymic GCTs confirmed at histopathology were enrolled retrospectively, and their CT and MRI findings were analysed. RESULTS: According to the CT findings, malignant thymic GCTs usually manifest as a bulky mass that typically grows to both sides of the midline (20/22, 90.9%), with irregular shape (15/22, 68.2%), lobulation (12/22, 50%), ill-defined margin (9/22, 40.9%), and incomplete capsule (21/22, 95.5%). Twenty masses revealed heterogeneous density with multifocal necrosis or cystic change in 19 (86.4%). Most cases (16/18, 88.9%) showed mild to moderate enhancement, and the branch-like vessel was found in 14 (14/18, 77.8%) cases. The minimum apparent diffusion coefficient (ADCmin) and mean apparent diffusion coefficient (ADCmean) values in 14 patients were (1.13 ± 0.45) and (1.37 ± 0.49) × 10-3 mm2/s, respectively. Compared with CT findings, the incidences of an incomplete capsule, heterogeneous signal, and necrotic or cystic change on MRI images occurred in all patients with malignant thymic GCT. In addition, peritumoural oedema was found in all 14 (100%) cases. CONCLUSION: MRI is superior to CT in showing incomplete capsule, peritumoural oedema, heterogeneous signal, and necrotic or cystic change of GCTs. Branch-like enhancement and multifocal necrosis may help the diagnosis of malignant thymic GCTs.

[An arthroscopic "inlay" Bristow procedure with suture button fixation: Surgical technique and radiology evaluation].

OBJECTIVE: To introduce an arthroscopic "inlay" Bristow procedure based on the Mortise-Tenon joint structure concept using suture button fixation, and to evaluate its clinical and radiology results postoperatively with a minimal 3-year follow-up. METHODS: A total of 56 patients who received arthroscopic "inlay" Bristow procedure with suture button fixation between June 2015 to June 2016 were eventually enrolled in this study. Radiological assessment on the 3D CT scan was performed preoperatively, immediately after operation, and postoperatively at the end of 3 months, 6 months and the final follow-up. Complications postoperatively were also recorded. RESULTS: A total of 56 patients were finally included in this study. The mean follow-up time was (36.1±3.7) months. Coracoid grafts (middle point) were positioned at about 4 o'clock (123.8°±12.3°) in the En-face view. In the axial view, 95% (53/56) of the grafts positioning were measured as flush, 5% (3/56) as medial. Bone union rate was 96.4% at the final follow-up. At the end of 3 months, 6 months, and the final follow-up, the length of the coracoid graft was 96.9%±4.9%, 91.9%±6.2%, and 91.6%±6.6% of the immediate postoperative length, respectively. Compared with the immediate postoperative length, the length measured at the end of 3 months shortened not significantly (t=2.12, P > 0.05). The coracoid graft shortened more pronouncedly 6 months postoperatively (t=4.98, P < 0.05) and then remained almost constant over time (t=-0.75, P > 0.05), with all grafted coracoid graft retaining more than 90% of their initial length by the 3-year follow-up. And new bone formation at the junction between the coracoid graft and glenoid neck in the axial view were obviously noted in 25 cases. The quantitative evaluation showed that the glenoid area in En-face view was significantly increased at the final follow-up than that immediately after surgery [(9.72±1.22) cm2 vs. (9.42±1.11) cm2]. No degenerative changes were noted on CT images in all the patients at the final follow-up. CONCLUSION: This study reported a series of "inlay" Bristow procedure with suture button fixation for recurrent shoulder dislocation, providing satisfactory union rate and excellent graft positioning. And using suture button fixation instead of screw can reduce osteolysis and complications related to hardware implantation. Moreover, the bone remodeling between the coracoid process and glenoid could be beneficial to restoring the anterior stability of shoulder joint in a long term follow-up.

[Two kinds of lateral retinacular release for lateral patellar compression syndrome:a retrospective comparative study].

Objective: To compare the clinical effects of arthroscopic lateral retinacular proximal release and "L" type release for lateral patellar compression syndrome. Methods: Sixty four lateral patellar compression syndrome (LPCS) patients were recruited who had arthroscopic surgery during September 2004 to January 2019 at Department of Sports Medicine,Peking University Third Hospital by the same group of doctors and a retrospective comparative study was conducted accordingly. Among them,24 cases underwent the traditional proximal release of lateral patellar retinaculum (traditional group),including 7 males and 17 females with an age of (42.9±14.6)years(range:23 to 72 years); 40 cases were treated with novel lateral retinacular release of "L" type release revised based on the previous experiences (novel group), consisting of 12 males and 28 females with an age of (54.9±13.1) years (range:28 to 76 years).All the patients participated surveys for the visual analogue scale (VAS),Lysholm score and International Knee Documentation Committee (IKDC) knee function subjective evaluation before and after the operation. The comparison of scores within groups was performed by Wilcoxon test,and comparison between groups was performed by Mann-Whitney U test and Chi-square test. Multivariate analysis was used to evaluate the related factors affecting each score. Results: The follow-up durations of traditional group and novel group were (12.6±1.0) years (range:11.7 to 15.3 years) and (2.2±1.0)years(range:1.0 to 4.4 years) respectively. At the last follow-up,VAS(0(1.0)to 6.0(2.0),Z=-5.471,P<0.01),Lysholm score(98.0(10.0)to 48.0(40.0),Z=-5.511,P<0.01),and IKDC score(82.8(11.2)to 37.4(18.5),Z=-5.444,P<0.01) in novel group were statistically significantly improved,and the postoperative excellent rate of Lysholm score was 97.5% in general. There was no significant difference in the changes of the three scores (P>0.05) between the traditional group and the novel group. However,20.8% (5/24)patients in the traditional group reported significant weakness of the knee extension after surgery,while no such complain was received in the novel group (P<0.01).The results of univariate analysis showed that surgical method was a related factor affecting the changes of VAS before and after surgery (P<0.05).The results of multivariate analysis showed that whether or not with osteoarthritis and operation type were independent factors affecting the changes of Lysholm and IKDC scores (P<0.05). Conclusions: The long-term effect of arthroscopic lateral retinacular release for the treatment of LPCS is satisfactory. Compared with the traditional proximal release surgery,the "L" type release can effectively avoid the complication of significant weakness of the knee extension significantly.

Action of Bacillus natto 16 on deoxynivalenol (DON) from wheat flour.

AIMS: The aim of this research is to study the removal characteristics and evaluate the detoxify action of deoxynivalenol by Bacillus natto 16 in wheat flour as food or feed. METHODS AND RESULTS: The content of deoxynivalenol was determined using ELISA by testing the deoxynivalenol removal rate, and the influence of culture supernatant, intracellular substances, crude enzyme and cell wall on the deoxynivalenol in wheat flour was studied. The effect of bacterial components on the removal of deoxynivalenol was studied in the artificial gastrointestinal environment to simulate the digestion of food. Secondary metabolites were analysed by high-performance liquid chromatography in tandem with mass spectrometry (HPLC-MS). The cell wall can reduce the content of deoxynivalenol in the sample by adsorption, the influence of culture supernatant, intracellular substances and crude enzyme can convert deoxynivalenol into substances with a lower molecular weight. Bacterial components have no effect on deoxynivalenol in wheat flour in simulated gastric fluid (SGF) and have a certain removal effect on deoxynivalenol, which is closely related to intestinal digestion time and pH, in simulated intestinal fluid. CONCLUSIONS: Experimental results indicate that the removal of deoxynivalenol by B. natto 16 includes adsorption and biodegradation, SGF would invalidate the deoxynivalenol removal activity of B. natto 16's components. SIGNIFICANCE AND IMPACT OF THE STUDY: Our study showed that as an edible probiotic bacterium, B. natto 16 can effectively remove deoxynivalenol from wheat flour as food or feed, and can be used as a new deoxynivalenol -detoxifying microbe. The results of this research could provide the theory foundation for further development and application of B. natto 16.

MiR-133b inhibits MPP+-induced apoptosis in Parkinson's disease model by inhibiting the ERK1/2 signaling pathway.

OBJECTIVE: The aim of this study was to explore the effect of micro ribonucleic acid (miR)-133b on 1-methyl-4-phenylpyridinium ion (MPP+)-induced apoptosis in the Parkinson's disease (PD) model. MATERIALS AND METHODS: PC12 cells were induced by different concentrations of MPP+ to establish the PD cell model. Subsequently, the survival rate of PC12 cells was detected using Cell Counting Kit-8 (CCK-8) assay. Quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR) was used to detect the expression of miR-133b in the PD model induced by different concentrations of MPP+. Next, PC12 cells were transfected with miR-133b mimic and miR-negative control (NC), and divided into MPP+ group, MPP+ + miR-NC group and MPP+ + miR-133b mimic group. Transfection efficiency was verified using qRT-PCR. The apoptosis of cells was detected using terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. Moreover, the expressions of extracellular signal-regulated kinase 1/2 (ERK1/2) and phosphorylated (p)-ERK1/2 were determined using Western blotting. RESULTS: After MPP+ treatment, the survival rate of PC12 cells significantly declined (p<0.05). MPP+ exhibited toxicity against PC12 cells in a concentration-dependent manner. Meanwhile, cell survival rate decreased remarkably with the increase of MPP+ concentration (p<0.05). With increased concentration of MPP+, the expression of miR-133b in the PD cell model declined significantly (p<0.05). The apoptosis of PC12 cells was remarkably inhibited by overexpression of miR-133b in the PD cell model (p<0.05). In addition, the protein expression of p-ERK1/2 in PC12 cells was notably reduced after overexpression of miR-133b in the PD cell model (p<0.05). CONCLUSIONS: MiR-133b is lowly expressed in the PD cell model. Furthermore, overexpression of miR-133b inhibits cell apoptosis in the PD cell model by regulating the ERK1/2 signaling pathway.

EZH2-mediated H3K27me3 enrichment on the lncRNA MEG3 promoter regulates the growth and metastasis of glioma cells by regulating miR-21-3p.

OBJECTIVE: Glioma is one of the most common and invasive brain tumors worldwide. Long non-coding RNAs (LncRNAs) play an important role in the development of glioma. However, the regulatory mechanism of LncRNAs in glioma has not been fully elucidated. This study aimed to explore the interaction of lncRNA maternally expressed gene 3 (MEG3) and aberrant histone modification in glioma. MATERIALS AND METHODS: The expression levels of MEG3 and miR-21-3p in glioma cells were measured by quantitative polymerase chain reaction (qPCR). EZH2 (enhancer of zeste homolog 2) and H3K27me3 expression in glioma cells were detected by Western Blot (WB). The binding site of the promoter of MEG3 by H3K27me3 was confirmed by ChIP-Real-time PCR. The direct target of MEG3 and miR-21-3p in glioma cells was measured by a luciferase reporter assay. Cell proliferation was detected by Cell Counting Kit-8 (CCK8), and cell invasion and migration were measured by Transwell assays. RESULTS: EZH2 and miR-21-3p were upregulated and MEG3 was downregulated in glioma cells. Silencing of EZH2 inhibited cell proliferation, migration, and invasion in U87 and U251 cells. Meanwhile, the expression of H3K27me3 could be significantly inhibited by EZH2 interference. H3K27me3 protein can bind to MEG3 promoter directly. EZH2 inhibition and MEG3 down-expression in U87 cells reversed the effects of silencing of EZH2 on glioma cell growth and metastasis. However, EZH2 inhibition and MEG3 overexpression in U251 cells restricted cell proliferation, migration, and invasion. Furthermore, miR-21-3p was verified to interact with MEG3 by direct binding. Inhibition of MEG3 promoted U87 cell growth and metastasis, which was further strengthened following the co-transfection of si-MEG3 and miR-21-3p. Overexpressed MEG3 inhibited U251 cell growth and metastasis and a complete reversal of the results observed in the co-transfection of LV-MEG3 and miR-21-3p. CONCLUSIONS: EZH2 was highly expressed in glioma cells and EZH2-mediated H3K27me3 enrichment on the MEG3 promoter regulated the growth and metastasis of glioma cells by targeting miR-21-3p. It potentially provided a new therapeutic marker targeting glioma.

[Studies on the role of chromobox protein homolog 2 in the inhibition of progression of hepatoma].

Objective: To explore chromobox protein homolog 2 (CBX2) expressions in relation to clinical features of patients and elucidate its role in the progression of hepatocellular carcinoma. Methods: Using the Cancer Genome Atlas (TCGA) database, R language was used to analyze the distribution of differentially expressed mRNA in hepatocellular carcinoma. The different expression of CBX2 in HCC and adjacent tissues and its relationship with survival and clinical characteristics of patients were further analyzed. The expression of CBX2 in liver tissues, liver cancer tissue, and L02, HepG2 and SMMC-7721 cell lines was detected by real time-PCR and western blot. The expression of CBX2 was interfered by siRNA in hepatoma cell line. MTT, colony formation, transwell assays, and flow cytometry were used to identify the proliferation, apoptosis, invasion and clone-formation ability of HepG2 and SMMC-7721 cells after CBX2 down-regulation. According to the different data, t-test, ANOVA, chi-square test, and COX regression model were used for statistical analysis. Survival curve was plotted through Kaplan-Meier method. Results: TCGA public database analysis showed that the expression of CBX2 mRNA in hepatocellular carcinoma tissues (7.296 ± 1.6115) was significantly higher than normal liver tissues (4.706 ± 0.940) (P = 0.000). In addition, the overall survival time of patients with low CBX2 mRNA expression was significantly longer than that of patients with high CBX2 mRNA expression [(5.971 ± 0.411) years vs. (4.650 ± 0.503) years, P = 0.001]. The expression level of CBX2 mRNA was correlated with the pathological TNM stage (P = 0.025) and differentiation degree (P < 0.001) of liver cancer. COX regression analysis showed that CBX2 mRNA expression was an independent predictor of patient survival (P = 0.013). siRNA was transfected and compared with the blank control group. The transgenic ability of HepG2 and SMMC-77221 cells decreased significantly at 72h (P < 0.05) and 96h (P < 0.05), and the apoptosis rate (11.430% ± 0.215%) was higher than blank control group (6.6 00% ± 0.170%) (P = 0.003). The number of invasive cells ((both P < 0.05) and relative colony forming cells ((both P < 0.001) were significantly decreased. In 20 cases of tissue samples, the expression of CBX2 protein (relative expression level 3.020 ± 0.269) in liver cancer was higher than that in adjacent tissues (relative expression level 0.886±0.065) (P < 0.001). The overall survival time of patients with low CBX2 expression in liver cancer was longer than that of patients with high expression [(3.670 + 0.576) years vs. (0.834 + 0.153) years, P = 0.004]. Conclusion: An evident high expression of CBX2 is an independent poor prognostic factor in hepatoma. Down-regulation of CBX2 expression can inhibit the progression of liver cancer. Therefore, CBX2 may be a prognostic biomarker and a new target for HCC treatment.

[Clinical and genetic manifestations of immunodeficiency, centromeric instability, and facial anomalies syndrome: a case report and literature review].

Objective: To analyze the clinical and genetic features of immunodeficiency, centromeric instability, and facial anomalies (ICF) syndrome with a case report and literature review. Methods: The clinical data and genetic test of a girl diagnosed with ICF syndrome in the Department of Nephrology and Immunology in Qingdao Women and Children's Hospital in December 2016 were extracted and analyzed. "ICF syndrome" "immunodeficiency, centromeric instability and facial anomalies syndrome" "ICF syndrome and DNMT3B" were used as key words to search Chinese databases and Pubmed for literature until March 2018, and the literature was reviewed. Results: A female patient aged 22 months old with ocular hypertelorism and low-set ears was admitted due to recurrent infection over one year. Laboratory tests showed humoral immune deficiency with IgG<1.34 g/L, IgA<0.060 g/L, and IgM<0.179 g/L, but normal cellular immunity (total T lymphocyte 0.503, hepler T lymphocyte 0.328, cytotoxic T lymphocyte 0.166, natural killer cell 0.184, total B lymphocyte 0.276). Whole-exome sequencing revealed a de novo heterozygous splice site mutation c.922-2A>G in intron 8, and a de novo heterozygous missense mutation c.2477G>A in exon 23 of DNMT3B gene. Chromosome karyotype analysis showed 46, XX, with 64 out of 100 karyotypes showing centromere instability in chromosome 1. Five papers were found which were all in English, with total of 29 patients. Forty-three mutations were reported, including 34 missense, 2 deletion, 1 insertion, 6 splice site mutations. Eleven patients had complex heterozygosis mutations. All patients had centromere instability, humoral immune deficiency and facial dysplasia which were mainly ocular hypertelorism and low-set ears. Most patients had language and motor development delay, and a few were combined with mental retardation. Conclusions: ICF syndrome is a rare autosomal recessive primary immunodeficiency with classic clinical triad manifestations. De novo mutation of DNMT3B gene is one of etiologies according to genetic test.

Diagnostic value of transvaginal four-dimensional hysterosalpingo-contrast sonography combined with recanalization in patients with tubal infertility.

BACKGROUND: A retrospective study was designed to evaluate the diagnostic value of transvaginal four-dimensional hysterosalpingo-contrast sonography (TVS 4D-HyCoSy) combined with recanalization versus laparoscopy for patients with tubal infertility. MATERIALS AND METHODS: A total of 195 patients undergoing TVS 4D-HyCoSy were analyzed retrospectively. Of these, 72 patients underwent laparoscopy, which was the gold standard. The endpoints were coincidence rate (defined as a parameter consistent with results arising from TVS 4D-HyCoSy and laparoscopic examination using dye), sensitivity, specificity, positive predictive value, negative predictive value, and Youden index for TVS 4D-HyCoSy. RESULTS: A total of 385 fallopian tubes were assessed by TVS 4D-HyCoSy, of which 147 (38.2%) were tubal patency, 178 (46.2%) as partial tubal obstruction, and 60 (15.6%) as complete tubal obstruction. Of 195 patients, 72 patients with 144 fallopian tubes underwent laparoscopy and a total coincidence rate of 90.97% compared with TVS 4D-HyCoSy. The sensitivity, specificity, positive predictive value, negative predictive value, and Youden index for 4D-HyCoSy versus laparoscopy were 97.7%, 86.7%, 98.4%, 81.3%, and 0.84, respectively. CONCLUSIONS: TVS 4D-HyCoSy represents a highly useful method for diagnosing tubal patency. However, further large-scale studies are warranted to investigate our findings in patients with tubal infertility.

[Clinicopathological features and treatment outcome of 37 cases of primary thyroid lymphoma].

Objective:The aim of this study is to explore the clinicopathological features and treatment outcome of primary thyroid lymphoma(PTL). Method:Clinicopathological data of 37 cases of PTL were retrospectively reviewed, and analyzed in combined with followup data. Result:Of the 37 patients, Hashimoto's disease was diagnosed in 28(75.7%) patients. The diagnostic rate of preoperative fine needle biopsy (75%) was higher than the fine needle aspiration (53%). The pathological types of these patients included diffuse large B cell lymphoma(n=23), extranodal marginal zone lymphoma of mucosaassociated lymphoid tissue type (MALT) (n=10), follicular lymphoma(FL) grade 3 (n=3) and burkitt lymphoma(BL) (n=1). According to Ann Arbor staging system, 12 patients with stage ⅠE, and 25 patients with stage ⅡE. With a median followup period of 37 months (2－93 months), the 3year overall survival (OS) rate and progression free survival(PFS) was 87.0% and 81.6%, and 5year overall survival(OS) rate and PFS was 79.8% and 74.8%. Univariate survival analysis found that comprehensive treatment was an important factor affecting prognosis, but there was no statistical difference in the effects of age, gender, lactate dehydrogenase, level, ß2 microglobulin, Ann Arbor staging, international prognostic index, and tissue subtype on survival (P>0.05). Conclusion:PTL mostly affects middle aged and old female and has a favorable prognosis. Core needle biopsy can improve the diagnostic rate when compared with fine needle aspiration. The optimal treatment is combined modality strategy based on chemotherapy and radiotherapy, and supplemented by surgery.

Research on changes in cognitive function, ß-amyloid peptide and neurotrophic factor in stroke patients.

OBJECTIVE: To investigate the changes as well as the related mechanism in cognitive function and levels of serum ß-amyloid peptide (Aß) and brain-derived neurotrophic factor (BDNF) in stroke patients. PATIENTS AND METHODS: A total of 30 patients with acute stroke treated in our hospital from June 2015 to September 2016 were selected as stroke group, while 30 volunteers during the same period were enrolled as control group. Changes in cognitive function of patients were evaluated using the Montreal Cognitive Assessment (MoCA) and mini-mental state examination (MMSE) before and after the treatment. At the same time, the concentrations of serum Aß1-40 and BDNF were detected, and their correlations with the MMSE score were analyzed. Finally, levels of serum cyclic adenosine monophosphate (cAMP) and phosphorylated-cAMP-response element binding protein (p-CREB), and the phosphorylation level of Tau protein were detected by Western blotting. RESULTS: MoCA and MMSE scores of patients in stroke group were significantly lower than those in control group (p < 0.01), and the scores were significantly higher in stroke patients after treatment than those before treatment (p < 0.01). Compared with those in control group, the serum Aß1-40 concentration in patients in stroke group was significantly increased (p < 0.01), but the BDNF level was significantly decreased (p < 0.01). Compared with those before treatment, the serum Aß1-40 concentration in patients was significantly decreased after treatment (p < 0.01), but the BDNF concentration was significantly increased (p < 0.01). Correlation analysis showed that the MMSE score was negatively correlated with the concentration of Aß1-40 (r2 = 0.764, p < 0.01), but positively related to the level of BDNF (r2 = 0.827, p < 0.01). Compared with those in control group, the content of serum cAMP and p-CREB in stroke patients was significantly decreased (p < 0.01), but the expression of p-Tau was statistically increased (p < 0.01). CONCLUSIONS: The cognitive function in stroke patients is impaired, with the rising content of serum Aß1-40 and reduction of BDNF, the mechanism of which is related to the decrease of cAMP and p-CREB and the increase of p-Tau. This provides a theoretical basis for searching the new therapeutic targets and new drugs for stroke.

G-quadruplex antibody attenuates human gastric cancer cell proliferation and promotes apoptosis through hTERT/telomerase pathway.

OBJECTIVE: To investigate the expression of G-quadruplex antibody BG4 in human gastric cancer AGS cells and assess its functions in attenuating proliferation and promoting apoptosis in gastric cancer. MATERIALS AND METHODS: BG4 high-expression gastric cancer AGS cell line was established by pEGFP-N1-BG4 transient transfection. AGS cells transfected with pEGFP-N1 plasmids were included into the pEGFP-N1 group and those not transfected with plasmids were included into the negative control group. Cell counting kit-8 (CCK8) assay was performed to examine the AGS cell proliferation ability, while flow cytometry was used to detect the cell cycle distribution and cell apoptosis. Cell migration was measured using Transwell migration and wound healing assay. Then the expression levels of cell apoptosis associated factors were determined. The mRNA and protein expressions of human telomerase reverse transcriptase (hTERT), B-cell lymphoma 2 (Bcl-2), Bcl-2 associated X (Bax) were examined with real-time quantitative polymerase chain reaction (PCR) and Western blotting, respectively. RESULTS: The results revealed that pEGFP-N1-BG4 group exhibited reduced proliferation and migration, induction of apoptosis. hTERT and Bcl-2 mRNA and protein levels in pEGFP-N1-BG4 group were down-regulated compared with those in the pEGFP-N1 group and control group, but there were no significant differences in Bax mRNA and protein levels compared with those in the pEGFP-N1 group and control group. CONCLUSIONS: We showed that the expression of BG4 in the gastric cancer cell line AGS inhibits cell proliferation and promotes apoptosis though inducing telomere to form G-quadruplex structure and attenuating telomerase activity, thus resulting in reduced expression of hTERT and Bcl-2.