Four-Hour Fasting for Semifluids and 2-Hour Fasting for Water Improves the Patient Experience of Esophagogastroduodenoscopy: A Randomized Controlled Trial

Background/Aims@#We aimed to investigate the comfort, safety, and endoscopic visibility during esophagogastroduodenoscopy (EGD) afforded by a modified 4-hour semifluid and 2-hour water (“4+2”) fasting protocol. @*Methods@#In this parallel group, endoscopist-blinded, randomized controlled trial, outpatients undergoing unsedated diagnostic EGD from 10:30 AM to 12:00 PM were randomly assigned to either a “4+2” protocol group or a conventional fasting group. The participants’ comfort during the fasting period and procedure was measured using the visual analog scale, and mucosal visibility was measured by endoscopists using the total visibility score. Satisfaction was defined as a visual analog scale score of ≤3. The primary outcome was the participants’ comfort during fasting. @*Results@#One hundred and six and 108 participants were randomized to the “4+2” protocol and control groups, respectively. Participants’ comfort before EGD was significantly higher in the “4+2” protocol group measured by both the proportion of satisfaction (86.8% vs 63.9%, p=0.002) and the visual analog scale score (median [interquartile range]: 1.0 [1.0–2.0] vs 3.0 [1.0–4.0], p<0.001). The proportion of satisfaction during EGD also significantly improved (59.4% vs 45.4%, p=0.039) in the “4+2” protocol group. The total visibility score was unaffected by the fasting protocol (5.0 [4.0–5.0] vs 4.0 [4.0–5.0], p=0.266). No adverse events were observed during the study. @*Conclusions@#The “4+2” protocol was more comfortable and provided equal mucosal visibility and safety compared with conventional fasting for unsedated EGD.

Key points in pancreatic cancer： ESMO clinical practice guideline for diagnosis， treatment and follow-up （2023） / 临床肝胆病杂志

In September 2023， the European Society for Medical Oncology published Pancreatic cancer： ESMO Clinical Practice Guideline for diagnosis， treatment and follow-up in Annals of Oncology. The guideline provides recommendations on the epidemiology， diagnosis， and treatment of pancreatic cancer （PC）. The guideline highlights the high-risk populations of PC and recommended screening methods， clarifies the diagnostic process of PC and various radiological and molecular biology tests， and mentions the appropriate treatment methods for different types of PC. This article makes an excerpt of this guideline and introduces related recommendations for the clinical treatment of PC， in order to provide a reference for clinical practice.

Application of endoclip- and submucosal injection-assisted cannulation technique to difficult cannulation during endoscopic retrograde cholangiopancreatography (with video) / 中华消化内镜杂志

Objective:To evaluate the value of endoclip-assisted and submucosal injection-assisted cannulation techniques for difficult cannulation during endoscopic retrograde cholangiopancreatography (ERCP).Methods:Data of 12 458 patients treated with ERCP for the first time in the First Affilated Hospital of Naval Medical University from June 2015 to September 2020 were retrospectively analyzed. Twenty eight (0.22%) were identified as difficult cannulation where metal clip- or submucosal injection-assisted cannulation was used. The selective cannulation success rate, intubation time and complication incidence of the two techniques in difficult cannulation patients were analyzed.Results:Difficult cannulation was performed in 18 males (64.3%) and 10 females (35.7%) with an age of 69.6±14.1 years assisted by metal clips or submucosal injection. Five cases (17.9%) were type Ⅱ, 5 cases (17.9%) type Ⅲ, and 18 cases (64.3%) type Ⅴ according to papilla classification. Sixteen patients (57.1%) received metal clip-assisted cannulation, and 12 cases (42.9%) submucosal injection-assisted cannulation. Twenty-five (89.3%) patients successfully underwent selective cannulation with the cannulation time of 9.9±4.3 min. One case (3.6%) of mild post-ERCP pancreatitis and 3 cases (10.7%) of post-ERCP hyperamylasemia occurred. No postoperative bleeding or perforation occurred. All patients were cured and discharged after conservative treatment.Conclusion:When selective cannulation is difficult due to poor papilla exposure or deflection, endoclip- or submucosal injection-assisted cannulation can effectively improve the successful selective cannulation rate during ERCP with low complication incidence, which is worth of clinical promotion.

Prevalence of diabetes mellitus among Uygur children in Hotan Prefecture of Xinjiang, China / 中国当代儿科杂志

OBJECTIVES@#To investigate the prevalence of diabetes mellitus (DM) among Uygur children in Hotan Prefecture of Xinjiang, China, as well as the factors influencing the development of DM.@*METHODS@#The cluster random sampling method was used to select 5 308 children, aged 4-18 years, from the middle and primary schools and kindergartens in Hotan Prefecture of Xinjiang. The survey methods included questionnaire survey and the measurement of height and weight. All subjects were tested for fasting fingertip blood glucose to investigate the prevalence of DM and impaired fasting glucose (IFG).@*RESULTS@#A total of 5 184 valid questionnaires were collected. Fourteen children (0.27%) were found to have DM, among whom 8 had type 1 DM, 2 had type 2 DM, and 4 had unclassified DM. Twenty-nine children (0.56%) were found to have IFG. There was no significant difference in the prevalence rate of DM and IFG between boys and girls (P>0.05). The prevalence rate of DM was 0.18% in the 4-<10 years group, 0.47% in the 10-<15 years group, and 0.07% in the 15-18 years group (P=0.072).The prevalence rate of IFG in the above three age groups was 0.18%, 0.94%, and 0.42%, respectively, with a significant difference among groups (P=0.007). The proportion of family history of DM and the proportion of overweight/obesity in children with DM were significantly higher than those in children without DM (P<0.05), while the proportion of children with DM who preferred coarse grains was significantly lower than that in children without DM (P<0.05).@*CONCLUSIONS@#The prevalence of DM and IFG in Uyghur children in Hotan Prefecture of Xinjiang is relatively low. There is no significant difference in the prevalence of DM among children of different genders or age groups, but the prevalence of IFG in children of different age groups is different. A family history of DM, overweight or obesity, and low intake of coarse grains might be associated with the development of DM.

Endoscopy in China: a national survey on the services, quality and safety of digestive endoscopy in 2021 / 中华消化内镜杂志

Objective:To evaluate the services, quality and safety of digestive endoscopy in China in 2020.Methods:Data of digestive endoscopy in 2020 collected by the national medical quality information network were included. After data quality evaluation, the basic information of digestive endoscopy centers in different types of hospitals, the diagnosis and treatment of digestive endoscopy, the process and outcome indicators of digestive endoscopy were analyzed and compared.Results:A total of 3 714 hospitals were included in this survey. The digestive endoscopy operations completed by each hospital was 3 562.5 (1 299.75, 8 426.75), the digestive endoscopists was 4 (2, 7), and the endoscopic operations completed per capita per year was 900 (500, 1 452). The detection rate of early gastrointestinal cancer was 17.46% (110 069/630 265). The success rates of cecal intubation under colonoscopy and selective intubation of endoscopic retrograde cholangiopancreatography were 95.43% (6 976 521/7 310 970) and 94.21% (121 666/129 149) respectively. The complete resection rate of endoscopic submucosal dissection was 92.68% (93 536/100 924). The incidence of serious complications related to digestive endoscopic operations [0.05‰ (1 316/26 499 108)] and mortality [0.003‰ (80/26 499 108)] remained at a low level.Conclusion:The quality and safety of digestive endoscopy in China in 2020 is improved, but there are still some problems, such as the shortage of digestive endoscopists, the poor ability of endoscopists in the diagnosis of early gastrointestinal cancer.

Current status of endoscopy department of county hospitals in China: a cross-sectional study / 中华消化内镜杂志

Objective:To investigate the current status of endoscopy department of county hospitals in China, and to improve the construction quality of the endoscopy department of county hospitals.Methods:County hospitals from 20 provinces, autonomous regions and municipalities in 2019 were investigated in this study through an online survey. Questions were about the department scale, basic information of the endoscopy center, clinical diagnosis and treatment ability, teaching and academic ability.Results:A total of 114 county hospitals were included. The department of gastroenterology in county hospitals owned 38.9 beds, 8.6 doctors and 6.4 nurses on average in 2019. The mean host and endoscope were 3.1 and 11.0 respectively in each center. The mean outpatient, discharge and endoscopy procedure were 22 thousand , 2.3 thousand and 7.8 thousand respectively. The referral rate of 53 (46.5%) hospitals to higher-level hospitals was less than 3%. In terms of teaching and academy, 5 (4.4%) in 114 hospitals had endoscopist training bases of Chinese Medical Association. Fifty-two (45.6%) county hospitals hosted academic conferences at the city level and above, 21 (18.4%) published SCI papers and 67 (58.8%) published papers in key Chinese journals.Conclusion:Endoscopy centers of most county hospitals are well constructed with comprehensive facilities, qualified medical ability, management, and a research team, which meets the recommendation level of service.

Current status of digestive endoscopy diagnosis and treatment in Shanghai / 中华消化内镜杂志

To investigate the current status of digestive endoscopy diagnosis and treatment in Shanghai, and to provide decision-making support for the revision of the diagnostic and treatment standards and quality control criteria of digestive endoscopy in Shanghai. A total of 145 hospitals providing digestive endoscopy service were investigated through an online survey. The main survey contents were the situation of digestive endoscopists, the development of diagnosis and treatment technology, and the quality and safety situation. There were 1 212 digestive endoscopists in Shanghai, accounting for 1.59% of China which ranked first in all registered practioners in 2019. The annual diagnostic and treatment procedures of digestive endoscopy was 1 902.6 thousand, and endoscopists' working pressure was relatively high. In terms of quality control indicator of a single disease, detection rates of early gastrointestinal cancers and adenomas remained at a high level. The number of digestive endoscopists, endoscopic procedures and quality control indicators of a single disease in Shanghai was in a leading position in China in 2019.

Comparative study on metastatic lymph node radio and pathological lymph node stage in prognosis evaluation of patients with gastric cancer after radical resection / 中华内分泌外科杂志

Objective:To study the value of metastatic lymph node radio (rN) and pathological lymph node stage (pN) in evaluating the prognosis of patients after radical gastric cancer.Methods:The clinicopathological data of 491 patients who underwent radical gastrectomy in Tantai Yantaishan Hospital from Jan. 2013 to Dec. 2017 were retrospectively analyzed. X-tile software was used to group the metastatic lymph node radio by rN. According to the number of lymph node metastasis, pN stage was performed. The correlation between metastatic lymph node radio and other clinicopathological factors was assessed. The metastatic lymph node radio and the pathological lymph node stage in evaluating the prognosis of patients after radical gastric cancer were compared.Results:(1) X-tile analysis showed that the best cut-off values for the metastatic lymph node radio in this study were 0.14 (14%) and 0.63 (63%) . (2) According to the cut-off value, the 491 patients included in the study were divided into rN1 (256 cases) , rN2 (160 cases) , and rN3 (75 cases) three subgroups. The results of the analysis of differences showed that there were significant differencesbetween the groups in terms of tumor diameter, tumor location, surgical resection range, stage, lauren classification, degree of differentiation, pT, pN, vascular cancer emboulus, nerve invasion, and pathological TNM staging groups. (3) Comparison of rN and pN staging in evaluation of the prognosis of patients after radical gastric cancer: ①Kaplan-Meier survival analysis results showed that rN was better than pN. ② Both single factor and multivariate cox analysis showed that rN was an independent risk factor for the prognosis of gastric cancer. In univariate analysis, rN group HR=3.18 (95% CI 2.63-3.84, P<0.001) , pN stage HR=1.88 (95% CI 1.66-2.15, P<0.001) ; rN group HR=2.21 in multivariate analysis (95% CI 1.73-2.82, P<0.001) , pN staging HR=1.31 (95% CI 0.95-1.79, P=0.095) . ③The time-dependent ROC analysis showed that the prognostic ability of rN was better than pN staging before 52 months of postoperative follow-up, and pN staging was more advantageous after 52 months. ④The Lauren classification was used as a stratification factor for stratified analysis. The Kaplan-Meier survival curve indicated that rN was better than pN staging in intestinal, mixed and diffuse gastric cancer, and the AUC curve showed the prediction of rN in patients with mixed and diffuse gastric cancer was better than pN staging, while pN staging performance was slightly better in patients with intestinal gastric cancer. Conclusions:rN is an independent factor affecting the prognosis of patients after radical gastric cancer surgery. When judging the prognosis of patients within 52 months after radical gastric cancer, rN has a better prognostic value than pN. In patients with mixed and diffuse gastric cancer in the Lauren classification, rN shows better prognostic value.

Endoscopic mucosal resection for gastrointestinal neuroendocrine tumors with diameter≤10mm: a retrospective analysis of 40 cases / 中华内分泌外科杂志

Objective:To investigate the safety and reliability of endoscopic mucosal resection (EMR) in the treatment of gastrointestinal neuroendocrine tumors (GI-NETs) with a diameter of ≤10mm.Methods:From February 2015 to June 2019, 40 cases of gastrointestinal neuroendocrine tumors with a diameter of ≤10mm underwent EMR in Yantaishan Hospital of Yantai City were retrospectively analyzed to evaluate the safety and reliability of EMR surgery.Results:All 40 cases were performed endoscopically by EMR. The en bloc resection rate and RO resection rate reached 100%. The lesions were all confined to the submucosa and did not infiltrate the muscularis propria. Postoperative pathological classification: NET G1: 35 cases, NET G2 grade: 5 cases, no NET G3 grade cases. There were no complications such as bleeding and perforation during and after the operation. Follow-up: 1 case was lost to follow-up, 1 patient died of other diseases, and the remaining 38 cases were followed up for an average of 27.8 months (5-46 months) without local recurrence or distant metastasis.Conclusion:For well-differentiated G1 and G2 gastrointestinal neuroendocrine tumors with a diameter ≤10mm, EMR is a safe and reliable treatment.

Objective Quantitation of EGFR Protein Levels using Quantitative Dot Blot Method for the Prognosis of Gastric Cancer Patients

Purpose@#An underlying factor for the failure of several clinical trials of anti-epidermal growth factor receptor (EGFR) therapies is the lack of an effective method to identify patients who overexpress EGFR protein. The quantitative dot blot method (QDB) was used to measure EGFR protein levels objectively, absolutely, and quantitatively. Its feasibility was evaluated for the prognosis of overall survival (OS) of patients with gastric cancer. @*Materials and Methods@#Slices of 2×5 μm from formalin-fixed paraffin-embedded gastric cancer specimens were used to extract total tissue lysates for QDB measurement. Absolutely quantitated EGFR protein levels were used for the Kaplan-Meier OS analysis. @*Results@#EGFR protein levels ranged from 0 to 772.6 pmol/g (n=246) for all gastric cancer patients. A poor correlation was observed between quantitated EGFR levels and immunohistochemistry scores with ρ=0.024 and P=0.717 in Spearman's correlation analysis. EGFR was identified as an independent negative prognostic biomarker for gastric cancer patients only through absolute quantitation, with a hazard ratio of 1.92 (95% confidence interval, 1.05–3.53; P=0.034) in multivariate Cox regression OS analysis. A cutoff of 208 pmol/g was proposed to stratify patients with a 3-year survival probability of 44% for patients with EGFR levels above the cutoff versus 68% for those below the cutoff based on KaplanMeier OS analysis (log rank test, P=0.002). @*Conclusions@#A QDB-based assay was developed for gastric cancer specimens to measure EGFR protein levels absolutely, quantitatively, and objectively. This assay should facilitate clinical trials aimed at evaluation of anti-EGFR therapies retrospectively and prospectively for gastric cancer.

Technical and Clinical Aspects of Diagnostic Single-Balloon Enteroscopy in the First Decade of Use: A Systematic Review and Meta-analysis

Background/Aims@#Single-balloon enteroscopy (SBE) has been widely used in diagnosing small bowel disease. We conducted this study to systematically appraise its technical and clinical performance. @*Methods@#Studies on SBE published by September 2018 were systematically searched. Technical and clinical performance data were collected and analyzed with descriptive or meta-analysis methods. @*Results@#In total, 54 articles incorporating 4,592 patients (6,036 procedures) were included. Regarding technical parameters, the pooled insertion depths (IDs) for anterograde and retrograde SBE were 209.2 cm and 98.1 cm, respectively. The pooled retrograde ID in Asian countries was significantly greater than that in Western countries (129.0 cm vs 81.1 cm, p<0.001). The pooled anterograde and retrograde procedure times were 57.6 minutes and 65.1 minutes, respectively.The total enteroscopy rate was 21.9%, with no significant difference between Asian and Western countries. Clinically, the pooled diagnostic yield of SBE was 62.3%. Obscure gastrointestinal bleeding (OGIB) was the most common indication (50.0%), with a diagnostic yield of 59.5%. Vascular lesions were the most common findings in Western OGIB patients (76.9%) but not in Asian ones (31.0%). The rates of severe and mild adverse events were 0.5% and 2.5%, respectively. @*Conclusions@#SBE is technically efficient and is clinically effective and safe, but total enteroscopy is relatively difficult to achieve with this technique. Etiologies of OGIB in Asian countries differ from those in Western countries.

Advances in endoscopic therapy for difficult common bile duct stones / 临床肝胆病杂志

Common bile duct stones (CBDSs) are a common biliary tract disease, and endoscopic therapy is the first-line treatment modality for this disease; however, some CBDSs are difficult to remove by conventional endoscopic techniques and are thus called difficult CBDSs. Therapies for difficult CBDSs include endoscopic papillary large balloon dilation, mechanical lithotripsy, choledochoscopy-assisted lithotripsy, and extracorporeal shockwave lithotripsy. This article reviews the advances in the above mentioned techniques for the treatment of difficult CBDSs, including indications, clinical effect, and adverse events.

Prognostic factors of esophageal cancer with multiple organ metastases and a prediction nomogram / 中国实用内科杂志

OBJECTIVE: To investigate the prognostic factors of esophageal cancer with multiple organ metastases and establish a prognostic prediction model. METHODS: Patients data were extracted from the SEER database. The clinical data of 388 patients with esophageal cancer with multiple organ metastases were retrospectively analyzed. Risk factors were analyzed by log-rank method and survival curves were drawn by K-M method. Multivariate analysis was performed by Cox proportional hazard model to obtain independent prognostic factors for multi-organ metastasis of esophageal cancer. A prediction nomogram was further established.RESULTS: The mean survival time of patients in this study was 7.3 months, and the survival rates for 1-, 3-, and 5-year were 15.5%,1.2%, and 0, respectively. Age was an independent prognostic factor. The value of C-index was 0.618. CONCLUSION: The prognosis of esophageal cancer with multiple organ metastases is poor. Age at the diagnosis and patterns of multiple organ metastases are related to the survival time of patients. The prediction nomogram provided a good prognosis prediction.

Nomogram: an effective prognostic model for esophageal cancer with lung metastasis / 中国实用内科杂志

OBJECTIVE: To establish a prediction model for the prognosis of patients with esophageal cancer lung metastasis.METHODS: Data from 194 patients with esophageal cancer lung metastasis from 2010 to 2015 was collected from the National Cancer Institute Surveillance, Epidemiology and End RESULTS:(SEER) database. The best cutoff value for age was determined by X-tile software.Prognostic factors were analyzed by SPSS(v25.0) with the log-rank method and the Cox proportional hazard model. Risk factors from univariate analysis were used to construct prediction nomogram with R studio software(version 3.5.1). RESULTS: The median survival time of 194 patients with esophageal cancer lung metastasis was 7.0 months, the 3-month survival rate was 69.9%, and the 1-year survival rate was 27.7%. Age(HR=1.51, 95% CI: 1.066-2.140) and pathological type(HR=0.736, 95% CI: 0.543-0.998) were independent prognostic factors for patients with esophageal cancer lung metastasis. The value of C-index was 0.634(95% CI=0.585-0.683). CONCLUSION: For patients with esophageal cancer lung metastasis, being young and adenocarcinoma are associated with a better prognosis. The prediction of the nomogram is good.

Establishment of a prognostic nomogram for esophageal cancer liver metastasis–a population-based analysis / 中国实用内科杂志

OBJECTIVE: To analyze the prognostic factors related to liver metastasis of esophageal cancer and establish an effective prediction model. METHODS: The data of 464 cases of esophageal cancer with liver metastasis from 2010 to 2015 was collected from the National Cancer Institute SEER database by SEER stat 8.3.5 software. SPSS(v25.0) was used to analyze the prognostic factors of esophageal cancer liver metastasis and Kaplan-Meier curve was used for survival analysis. We introduced the meaningful variables of single factor analysis in Cox proportional hazard model and multivariate analysis and obtained the independent influencing factors of prognosis.Independent factors were then included in the accelerated failure time model to construct the nomogram. RESULTS: The mean survival time of patients in this study was 11.6 months(95%CI: 10.075-13.209), and their 1-, 3-, and 5-year survival rates were 29.4%, 5.5%, and 0,respectively. Age(HR=1.452, 95% CI: 1.175-1.795), marriage(HR=0.753, 95%CI: 0.611-0.927) and surgery(HR=0.428, 95% CI: 0.227-0.807) were independent prognostic factors for patients. We constructed the nomogram with risk factors of prognosis, and the C-index value was 0.614. CONCLUSION: The prognosis of esophageal cancer liver metastasis is poor. being young, Being married, and surgery are associated with better survival, and the nomogram we have constructed is proved to have good predictive ability.

Prognostic factors analysis and establishment of prediction model for 183 cases of esophageal cancer bone metastasis / 中国实用内科杂志

OBJECTIVE: To construct a model for predicting the prognosis of esophageal cancer bone metastasis. METHODS: The clinical data of 183 patients with esophageal cancer bone metastasis were analyzed retrospectively, and the prognostic factors of patients were analyzed by log-rank method and Cox proportional hazard model. Nomogram was constructed with the accelerated failure-time model.RESULTS: The average survival time(10.0 months, 95% CI:7.758-12.338) of patients aged 28-70 years was longer than that of patients aged 71-91(6.4 months, 95% CI:4.365-8.428)(χ~2=4.077, P=0.043). The prognosis of unmarried patients(average 7.0 months) was worse than that of the married(10.5 months on average)(χ~2=12.841, P<0.001). As for prognoses of different pathological types, the differences between adenocarcinoma(average 10.2 months, 95% CI:7.797-12.548), squamous cell carcinoma(average 6.4 months,95%CI:3.895-8.899) and other types(average 4.0 months, 95% CI:4.000-4.000) were statistically significant(χ~2=7.171, P=0.028).There were also significant differences between the prognoses of patients with different T stage(χ~2=8.833, P=0.032). Nomogram was constructed with the risk factors above and the C-index reached 0.675(95%CI: 0.626-0.725). CONCLUSION: The prognosis of esophageal cancer bone metastasis was poor. Marriage, T stage, histological grade and pathological types were risk factors affecting prognosis, while N stage didn't appear to show obvious effect on prognosis. The nomogram was tested to have a good predictive capacity.

Prognosis analysis of brain metastasis from esophageal cancer / 中国实用内科杂志

OBJECTIVE: To investigate the prognostic factors of esophageal cancer with brain metastasis. METHODS: SEER Stat 8.3.5 was used to collect 39 cases of esophageal cancer with brain metastasis from 2010 to 2015 in the Surveillance, Epidemiology and End RESULTS:(SEER) database. X-tile software was used to determine the best cut-off value of the age. Prognostic factors were analyzed with log-rank and Cox proportional hazard model by SPSS(v25.0). RESULTS: The median survival time of patients with esophageal cancer with brain metastasis was 7.0 months, the 6-month survival rate was 53.3%, and the 1-year survival rate was 16.3%. Only age(χ~2=4.045, P=0.044)was the prognostic factor, while there was insufficient evidence to show whether gender, marriage, race, primary site, histological grade,surgery, pathological type, T stage or N stage was associated with the prognosis of the patients. CONCLUSION: Brain metastasis is a rare metastatic type of esophageal cancer. Age is associated with worse prognosis, while the influences of other risk factors are not clear.Active treatment can lead to better prognosis.

Advances in research and clinical application of tumor markers in esophageal cancer / 中国实用内科杂志

Esophageal cancer is a common malignant tumor of the upper gastrointestinal tract. Early symptoms of the disease are inconspicuous and the disease is often diagnosed at a later stage, leading to higher morbidity and mortality. Esophageal cancer morbidity and mortality in both genders ranks among the top 10 most common cancers. Early detection and early treatment are effective means to reduce the incidence and mortality of esophageal cancer. Tumor markers play an important role in early diagnosis, treatment monitoring and prognosis evaluation of esophageal cancer. This paper reviews the clinical application of tumor markers related to esophageal cancer and the exploration and application progress of new tumor markers for esophageal cancer.

Effects of diallyl trisulfide in human fibroblast-like synoviocytes from rheumatoid arthritis / 中华风湿病学杂志

Objective To study the proliferation and inflammatory phenotypes of human fibroblast-like synoviocytes (FLS) induced by tumor necrosis factor-α(TNF-α). Methods The rheumatoid arthritis (RA)-FLS were cultured in vitro, then treated with different concentrations of diallyl trisulfide (DATS). The proliferation activity was detected by CCK-8 method. Then TNF-α was used to stimulate the RA-FLS, mRNA and protein expression of interleukin (IL)-6, matrix metalloproteinases (MMP)-1 and vascular endothelial growth factor (VEGF) were detected by quantitative real time polymerase chain reaction (qPCR) and enzyme linked immunosorbent assay (ELISA). Differences among groups were determined by one-way analysis of variance (ANOVA), LSD-t test was used for comparison between 2 groups. Results RA-FLS was successfully is-olated and cultured in vitro. The positive rate of CD90 and CD29 in the RA-FLS was more than 90％. The proli-feration activity of RA-FLS treated with 100 μmoL/L, 200 μmol/L and 300 μmol/L DATS was (98.92 ± 0.40)％, (95.91±0.32)％, (94.05±0.24)％, respectively. As Compared with the normal control group, the pro-liferation activity of RA-FLS was lower, and the statistically significant difference is between normal control group 200 μmol/L and 300 μmol/L DATS (t=-4.46, P<0.05; t=-7.98, P<0.05). After TNF-α stimulation, the expression of IL-6's mRNA in experiment group (100μmol/L DATS) is higher compared with the model control group (t=5.74, P<0.05), but the change of IL-6's protein is no significant difference (t=-0.49, P=0.627). The differences of mRNA and protein expression levels of MMP-1 and VEGF between the experiment group (100μmol/L DATS) and the model control group were not statistically significant. The relative mRNA level [(0.42 ± 0.06), t=-23.47, P<0.05;(0.14±0.039), t=-36.59, P<0.05;(0.36±0.09), t=-13.1, P<0.05)] and the protein levels [(108.0±4.7) ng/L, t=-63.79, P<0.05, (26.0±1.0) ng/L, t=-9.68, P<0.05;(57.9±0.7), t=-34.59, P<0.05] of IL-6, MMP-1, VEGF in experiment group (200μmol/L DATS) were significantly decreased. And the relative mRNA level [(0.041 ±0.027), t=-38.48, P<0.05; (0.027 ±0.027), t=-41.22, P<0.05; (0.131 ±0.047), t=-17.74, P<0.05] and the protein levels [(24.2 ±2.3) ng/L , t=-88.69, P<0.05; (22.7 ±1.0) ng/L , t=-14.13, P<0.05; (34.5 ±1.7), t=-48.45, P<0.05] of IL-6, MMP-1, VEGF in the experiment group (300 μmol/L DATS) were also significantly decreased. The difference between the two groups was significant (t=-24.89, P<0.05; t=-4.45, P<0.05; t=-13.87, P<0.05). Conclusion DATS can inhibit the proliferation and the effect of TNF-αinduced secretion of IL-6, MMP-1 and VEGF in RA-FLS. The effect is dose-dependent.

Effects of ring finger protein 43 on matrix metalloproteinases secreted by fibroblast-like synoviocytes from rheumatoid arthritis / 中华风湿病学杂志

Objective To explore the effects of ring finger protein 43 (RNF43) on fibroblast-like synoviocytes (FLS) from patients with rheumatoid arthritis (RA).Methods Synovial tissues from patients with RA treated by knee arthroplasty were used to isolate FLSs by type 2 collagenase.RNF43 lentivirus overexpressing plasmid was constructed and transfected in to RA-FLS.After successful transfection,RNA and super natant of RA-FLS were extracted.QRT-polymerase chain reaction (PCR) and enzyme linked immunosorbent assay (ELISA) were used to detect the mRNA and protein expression levels of matrix metalloproteinase (MMP)-1,MMP-3 and MMP-13.Data were analyzed with Student's t test.Results Transfection efficiency could meet the test requirements when the multiplicity of infection was 40 and was in conjunction with appropriate concentration of polybrene.The mRNA of RNF43 increased for 26158-fold than the control group.In vitro,compared with the control group,RNF43 could significantly inhibit the mRNA of MMP-1,MMP-3 and MMP-13 and MMP-13 [(0.19±0.06),t=28.314,P＜0.05;(0.28±0.07),t=23.413,P＜0.05;(0.21±0.09),t=18.365,P＜0.05]and the protein of MMP-1,MMP-3 and MMP-13 and MMP-13 [(31.0±9.4) pg/ml,(17.1±2.1) pg/ml,t=3.198,P=0.029],MMP-3 [(38.7±8.1) pg/ml,(24.9±3.5) pg/ml,t=3.514,P=0.015],MMP-13 [(35.9±5.4) pg/ml,(20.6±2.9) pg/ml,t=5.632,P=0.001].Conclusion The results of study suggest that RNF43 could inhibit the secretion of MMPs in RA-FLS by suppressing the activity of Wnt signal pathway.