METTL3 and METTL14 are two components that form the core heterodimer of the main RNA m6A methyltransferase complex (MTC) that installs m6A. Surprisingly, depletion of METTL3 or METTL14 displayed distinct effects on stemness maintenance of mouse embryonic stem cell (mESC). While comparable global hypo-methylation in RNA m6A was observed in Mettl3 or Mettl14 knockout mESCs, respectively. Mettl14 knockout led to a globally decreased nascent RNA synthesis, whereas Mettl3 depletion resulted in transcription upregulation, suggesting that METTL14 might possess an m6A-independent role in gene regulation. We found that METTL14 colocalizes with the repressive H3K27me3 modification. Mechanistically, METTL14, but not METTL3, binds H3K27me3 and recruits KDM6B to induce H3K27me3 demethylation independent of METTL3. Depletion of METTL14 thus led to a global increase in H3K27me3 level along with a global gene suppression. The effects of METTL14 on regulation of H3K27me3 is essential for the transition from self-renewal to differentiation of mESCs. This work reveals a regulatory mechanism on heterochromatin by METTL14 in a manner distinct from METTL3 and independently of m6A, and critically impacts transcriptional regulation, stemness maintenance, and differentiation of mESCs.
Animals ; Mice ; Methylation ; Chromatin ; Histones/metabolism* ; RNA, Messenger/genetics* ; Methyltransferases/metabolism* ; RNA/metabolism*

The internal organs and meridians were associated with Yin and Yang, five elements, six qi, and time and space, based on the holistic view of heaven, earth and human, according to Huangdi Neijing. The syndrome differentiation system of six meridians and Zang Fu meridians were established by Shanghan Zabing Lun, on the basis of the three Yin, three Yang, six meridians, and five Zang system in Huangdi Neijing. We put forward the concept of the six meridians syndrome differentiation system of circular motion, considering that the six meridians syndrome differentiation system actually implies the theory of circular motion. The syndrome differentiation system was constructed with the circular model of one qi circulating around the road, rising left and falling right, corresponding up and down, and maintaining conservation in the middle as the core, integrating Yin and Yang, five elements, six qi, Zang Fu and meridians, qi, blood and body fluid, and the integration of heaven, earth and human, focusing on "disease location and disease nature", taking classical prescriptions as the main treatments, and cooperating with external treatments such as acupuncture and moxibustion. We organically combined the circular motion with the syndrome differentiation of the six meridians, systematically interpreted the physiological bases, pathological changes, progressive patterns, and the treatments, based on syndrome differentiation, by inheriting the classical thinking mode of Hetu, Luoshu,Zhouyi, Huangdi Neijing, ShennongHerbal Classic, and Shanghan Zabing Lun.

[Abstract] Objective ： ：To investigate the long-chain noncoding RNA (Lnc RNA) PCGEM1 regulating the lung cancer (LC) cell invasion and metastasis through the TGF-β/Smad signaling pathways. Methods： ：From March 2016 to May 2018, total 62 cases of LC patients receiving surgical treatment in our hospital were collected, including cancer tissues and normal tissues more than 2 cm away from the cancer tissues. qRT-PCR was used to detect the expression of lncRNA PCGEM1 and miR-148a in LC, corresponding para-cancer tissues and different LC cell strains. LncRNA PCGEM1 silenced cell line A549-siPCGEM1 and negative control A549-NC were constructed, and A549 was used as blank control. MTT and plate cloning assay were used to detect the effect of PCGEM1 on the proliferation of A549 cells. Transwell and scratch assay were used to detect the effect of PCGEM1 on the invasion and migration of A549 cells. The bioinformatics website StarBase was used to predict the complementary binding miRNAof PCGEM1. Furthermore, according to the website Targetscan, the genes that the corresponding miRNAs could target and bind were predicted. Results： ：qRT-PCR results showed that the expression of PCGEM1 in LC tissues and lung cancer cell lines was higher than that in normal tissues, and the expression level of miR-148a was lower than that in normal tissues (all P<0.05). The expression level of PCGEM1 in A549 cells was the highest, and the difference was statistically significant compared with other cell lines (P<0.05). After successful construction of PCGEM1 silenced cells, compared with the blank control group and A549-NC group, the cell OD492nm value of A549-siPCGEM1 group was significantly decreased, the number of cell clones and the number of matrigel matrix gels was significantly reduced, the cell migration rate was significantly reduced, the differences were statistically significant (P<0.05). According to the prediction results of StarBase website, PCGEM1 could be complementary to miR-148a, and the prediction analysis on microRNA.org website shows that miR-148a had a targeted binding site with TGF-β2. qRT-PCR and Western blotting results showed that the expression of miR-148a was significantly increased in the A549-siPCGEM1 group compared with the blank control group and A549-NC group, and the expression of TGF-β2 and p-Smad 2 was significantly decreased (P<0.05), while the expression of the above indicators in the blank control group and A549-NC group was not statistically significant (P>0.05). Conclusion： ：Lnc RNA PCGEM1 is highly expressed in lung cancer. High expression of PCGEM1 may enhance the TGF-β2/Smad2 signaling pathway by downregulation of miR-148a, thus promoting the development of LC and the malignant biological behavior.

Objective To analyze the pathological results and current treatment situation of patients with unexpected gallbladder carcinoma from multi-centers in China,and explore the diagnosis and treatment of unexpected gallbladder carcinoma.Methods The retrospective cross-sectional study was conducted.The clinicopathological data of 223 patients with unexpected gallbladder carcinoma who were admitted to the 8 clinical centers from January 2010 to December 2016 were collected,including 86 in the First Affiliated Hospital of Xi'an Jiaotong University,41 in the First Affiliated Hospital of Zhengzhou University,30 in the Eastern Hepatobiliary Surgery Hospital of the Second Military Medical University,27 in the Xinhua Hospital of Shanghai Jiaotong University,13 in the First Affiliated Hospital of Dalian Medical University,11 in the Tianjin Medical University Cancer Institute & Hospital,9 in the First Affiliated Hospital of Army Medical University (Third Military Medical University) and 6 in the Affiliated Hospital of North Sichuan Medical College.Treatment of patients with unexpected gallbladder carcinoma who were diagnosed by intraoperative frozen section biopsy and postoperative pathological examination followed guideline for the diagnosis and treatment of gallbladder carcinoma (2015 edition).According to tumor staging and patients' decision,postoperative adjuvant treatment was selectively performed.Observation indicators:(1) diagnosis and treatment of unexpected gallbladder carcinoma;(2) followup and survival.Follow-up using outpatient examination and telephone interview was performed to detect postoperative survival up to June 2017.Measurement data with normal distribution were represented as (x)±s.Measurement data with skewed distribution were described as M (range).The survival time was calculated using the Kaplan-Meier method.Results (1) Diagnosis and treatment of unexpected gallbladder carcinoma:of 223 patients with unexpected gallbladder carcinoma,80 were initially diagnosed using intraoperative frozen section biopsy [20 received T stage results (intraoperative T stage of 14 patients had not matched postoperative results),and 60 didn't receive T stage results],and 143 were initially diagnosed using postoperative pathological examination (13 were initially diagnosed with gallbladder benign disease by intraoperative frozen section biopsy and 130 didn't intraoperatively receive frozen section biopsy).Of 223 patients,209,10,3 and 1 were respectively confirmed as adenocarcinoma,adenoma canceration,neuroendocrine tumor and squamous cell carcinoma;6,16,32,73,75,12 and 9 were respectively detected in Tis,T1a,T1b,T2,T3 and T4 stages and undefined stage;140 underwent reoperations,including 106 with radical resection of gallbladder carcinoma and 34 with extended radical resection of gallbladder carcinoma;operation of 126 patients reached the standard and operation of 97 patients didn't reach the standard.Of 27 patients with postoperative complications,12 with postoperative hemorrhage received successful hemostasis by reoperations (7 with cystic artery hemorrhage and 5 with blood oozing from gallbladder bed);8 with suppurative cholangitis received endoscopic retrograde cholangiopancreatography and choledochotomy with drainage,including 2 deaths and 6 with improvement;2 with common bile duct injury were improved by reoperation of choledochojejunostomy + T tube drainage;2 were complicated with bile leakage induced to peritonitis and underwent bile duct repair with drainage,including 1 death and 1 with improvement;2 with hepatic failure died of treatment failure;1 with colonic injury was improved by reoperation of anastomosis.Of 223 patients,207 didn't receive postoperative adjuvant treatment and 16 received postoperative adjuvant treatment,including 8 with chemotherapy,4 with radiotherapy,2 with immunologic therapy and 2 with Chinese medicine treatment.(2) Follow-up and survival:of 223 patients,193 were followed up for 6-90 months,with a median time of 33 months.Of 193 patients with follow-up:① The operation of 2 patients in stage Tis reached the standard,including 1 with cholecystectomy and 1 with radical resection of gallbladder carcinoma,and the postoperative survival time of them were respectively 28 months and 52 months.② The operation of 14 patients in stage T1a reached the standard,including 8 with cholecystectomy and 6 with radical resection of gallbladder carcinoma,and the postoperative survival time of them were respectively (74±5)months and (79±6)months.③ Of 26 patients in stage T1b,13 and 13 received respectively cholecystectomy and radical resection of gallbladder carcinoma (reaching the standard),and postoperative survival time of them were respectively (66±4)months and (76±8)months.④ Of 68 patients in stage T2,25,37,4 and 2 patients received respectively cholecystectomy,radical resection of gallbladder carcinoma (reaching the standard),extended radical resection of gallbladder carcinoma (reaching the standard) and palliative resection,and postoperative survival time of them were respectively (42±7) months,(66±6) months,(42±3) months and (26±3) months.⑤ Of 71 patients in stage T3,20,48 and 3 patients received respectively cholecystectomy,radical resection of gallbladder carcinoma (reaching the standard) and extended radical resection of gallbladder carcinoma (reaching the standard),and postoperative survival time of them were respectively (39±8) months,(48± 11) months and (10±6) months.⑥ Of 12 patients in stage T4,3,1,5 and 3 patients received respectively cholecystectomy,radical resection of gallbladder carcinoma (reaching the standard),extended radical resection of gallbladder carcinoma (reaching the standard) and palliative resection,and postoperative survival time of them were respectively (10±4) months,12 months,(9± 5) months and (11±3) months.Conclusions The intraoperative frozen section biopsy and pathological results are the key points for diagnosis and treatment of unexpected gallbladder carcinoma.Patients in stage Tis and T1a should undergo cholecystectomy,while patients in stage T1b and above should undergo radical resection of gallbladder carcinoma or extended radical resection of gallbladder carcinoma.

Objective To assess the predictive value for survival of BioCliM index in liver cirrhosis caused esophageal and gastric varices bleeding(EGVB)treated by endoscopic variceal ligation(EVL),endoscopic injection sclerotherapy(EIS)and endoscopic tissue adhesives(ETA).Methods From December 2006 to December 2011, the clinical data of 166 hospitalized patients with first occurrence of EGVB caused by liver cirrhosis and received endoscopic therapies were retrospectively analyzed.The scores of model for end-stage liver disease(MELD),model for end-stage liver disease-Na(MELD-Na),BioCliM index and Child-Turcotte-Pugh(CTP)were calculated. Receiver operating characteristic(ROC)curve and area under the curve(AUC)were applied to assess the accuracy of the four models in one-year and two-year prognosis evaluation,and to obtain the best critical value,and the mortality rates were compared among groups.Chi-square test,t test and rank-sum test were performed for statistical analysis.Results Among 166 patients,the levels of creatinine,bilirubin,albumin,sodium,international normalized ratio and prothrombin time were(0.10 ± 0.06)mmol/L,(0.02 ± 0.01)mmol/L,(30.13 ± 5.06)g/L, (139.13 ± 4.27)mmol/L,1.50 ± 0.32 and(17.83 ± 2.88)s,respectively.During the one-year and two-year follow-up,there were 14 patients and 23 patients dead,respectively.During the one-year and two-year follow-up, the incidences of portal thrombosis of death group were lower than those of survival group(10/14 vs 93.4%,142/152;73.9%,17/23 vs 94.4%,135/143);and the differences were statistically significant(χ2=8.029 and 10.774, both P<0.01).During the one-year and two-year follow-up,BioCliM indexes of death group were 0.12(-0.82, 1.44)and -0.81(-0.87,0.92),respectively,which were both higher than those of survival group(-0.84,-0.94 to -0.73;and -0.84,-0.94 to -0.72),and the differences were statistically significant(Z= -3.074 and -2.260,both P<0.05).During the one-year follow-up,the AUC values of MELD,MELD-Na,BioCliM index and CTP score were 0.698,0.691,0.749 and 0.723,respectively.During the two-year follow-up,the AUC values of MELD,MELD-Na,BioCliM index and CTP score were 0.587,0.582,0.647 and 0.633,respectively. But there was no statistically significant difference in AUC between BioCliM index and MELD,MELD-Na,and CTP score in one-year and two-year follow-up for prognosis evaluation(Z=0.509,0.566,0.271,0.687,0.731 and 0.162,respectively;all P>0.05).The best critical value of BioCliM index was -0.234.Followed up for one year and two years,the mortality rates of patients with BioCliM index over -0.230 were higher than that of patients with BioCliM index less than -0.234(31.0%,9/29 vs 3.6%,5/137;34.5%,10/29 vs 9.5%,13/137);and the differences were statistically significant(χ2=23.242 and 12.526,both P<0.01).Conclusions BioCliM index has a high accuracy in one-year and two-year mortality rate evaluation in liver cirrhosis patients with EGVB and received endoscopical therapies.We should pay attention to the prognosis evaluation before the endoscopical therapy.

Objective: To explore the prognosis of patients with T1b stage gallbladder carcinoma underwent different surgical procedure. Methods: The clinicopathological data of 97 patients with T1b stage gallbladder carcinoma came from 8 clinical centers from January 2010 to December 2016 and 794 patients who were admitted to the SEER database of USA from January 1973 to December 2014 were analyzed.There were 891 patients including 254 males and 637 females (1.0∶2.5) with age of (69.5±12.0)years. There were 380 patients who were less than 70 years old, 511 patients who were more than 70 years old. And there were 213 patients with the diameter of tumor less than 20 mm, 270 patients with the diameter of tumor more than 20 mm, 408 patients were unclear. There were 196 patients with well differentiation, 407 patients with moderately differentiation, 173 patients with poorly differentiation, 8 patients with undifferentiated, 107 patients were unclear. In the 891 patients with T1b stage gallbladder carcinoma, there were 562 cases accepted the simple cholecystectomy, 231 cases with simple cholecystectomy plus lymphadenectomy, and 98 cases with radical cholecystectomy. The time of follow-up were until June 2017. χ² test was used to analyze the enumeration data, rank-sum test was used to analyze the measurement data, the analyses of prognostic factors were used Cox proportional hazards model, the survival analysis was performed using Kaplan-Meier method. Results: The results of Cox proportional hazards model indicated, age, differentiation, surgical procedure were the risk factors of prognostic(1.929(1.594-2.336), P<0.01; 1.842(1.404-2.416), P<0.01; 1.216(0.962-1.538), P<0.01). The results of Kaplan Meier test indicated, the overall survival of T1b stage gallbladder carcinoma were (85.5±3.8)months, the overall survival of patients with simple cholecystectomy were (71.3±4.4)months, the overall survival of patients with cholecystectomy plus lymphadenectomy were(87.6±5.8)months, and the overall survival of patients with radical cholecystectomy were(101.7±9.3)months. The overall survival of patients with cholecystectomy plus lymphadenectomy and radical cholecystectomy were more than simple cholecystectomy(P<0.05). There were 329 patients with Lymph nodes examined in and after operations(231 patients with cholecystectomy plus lymphadenectomy, 98 patients with radical cholecystectomy). There were 265 patients with negative lymph node metastasis, the overall survival were(98.3±4.2)months. There were 64 patients with positive lymph node metastasis, the overall survival were(75.5±3.1)months. The overall survival of 38 patients with cholecystectomy plus lymphadenectomy were(62.7±2.6) months, and 26 patients with radical cholecystectomy were (82.2±3.7)months. The overall survival of patients with radical cholecystectomy were more than cholecystectomy plus lymphadenectomy(P<0.05). Conclusions The T1b stage gallbladder carcinoma patients with cholecystectomy plus lymphadenectomy or radical cholecystectomy has improved the prognosis comparing with simple cholecystectomy, significantly. When lymph node metastasis occurs, radical cholecystectomy has improved the prognosis comparing with cholecystectomy plus lymphadenectomy.

Objective To explore the distribution and antimicrobial resistance of pathogens causing healthcare-associated infection(HAI) in a hospital,and provide theoretical basis for rational use of antimicrobial agents and control of HAL Methods HAI occurred in patients in this hospital between 2011 and 2015 were chosen,distribution of major pathogens causing HAI and antimicrobial resistance were analyzed.Results A total of 852 pathogenic strains were isolated,including 458 (53.75 ％) strains of gram-negative bacteria,259 (30.40 ％) fungi,and 135 (15.85 ％) gram-positive bacteria,the top 4 pathogens were Klebsiella pneumoniae (n =159,18.66 ％),Candida albicans (n =119,13.97％),Acinetobacter baumannii (n =81,9.51％),and Escherichia coli (n =74,8.69 ％).Isolation rate of gram-negative bacteria increased from 47.41 ％ in 2012 to 61.57％ in 2015,fungi increased from 24.78％ in 2011 to 34.45％ in 2014 (2015 was 27.95％),gram-positive bacteria decreased from 26.55％ in 2011 to 10.48％ in 2015.Resistance rates of Escherichia coli and Klebsiella pneumoniae to imipenem and meropenem were 8.11％-12.58％;Acinetobacter baumannii had the lowest resistance rate to cefoperazone/sulbactam(23.46％),resistance rate of Candida albicans to fluconazole was up to 59.66％,vancomycin-resistant gram-positive bacteria was not found.Conclusion The major pathogens causing HAI are opportunistic pathogens,antimicrobial resistance is serious,the species and distribution varies with different years,it is necessary to enhance specimen detection rate,strengthen the monitoring on pathogen resistance,and use antimicrobial agents rationally.

Objective To evaluate the safety and effectiveness of three endoscopic methods for re-moval of common bile duct stones (CBDs)accompanied with periampullary diverticula(PAD).Methods A total of 154 patients hospitalized at Taizhou Hospital and Taizhou No.1 People′s Hospital of Zhejiang prov-ince from December 2012 to July 2013 were divided randomly into three groups,i.e.,EST,EPBD and ES-BD group,and received the treatment of EST,EPBD and limited EST plus EPBD (ESBD)to extract CBDs, respectively.After 12 months of follow-up,the rate of full stone clearance,stones clearance rate in one time,the rate of mechanical lithotripsy,the rate of urgent lithotripsy,the average procedures,the average removal time and the complication incidence among three groups were compared.Results The rates of stone clearance in one time in group ESBD was higher than those of group EST and group EPBD (94.12% VS 78.43%,73.08%;P <0.05)with significant difference.The average procedures in group ESBD was lower than that of group EPBD (1.08 VS 1.31,P <0.05),which also showed significant difference.The occur-rence rates of early complication in group ESBD was lower than that of group EPBD (15.69% VS 34.61%, P <0.05).The occurrence rates of post-ERCP hyperamylasemia in group ESBD was lower than that of group EPBD (5.88% VS 21.15%)with significant difference (P <0.05).The incidence of pneumobilia in group EST was higher than those of group EPBD (52.27% VS 26.19%,P =0.013)and group ESBD (52.27%VS 27.66%,P =0.016).Conclusion The stone extraction efficiency of ESBD is better than that of EST and EPBD.Compared with conventional EST,ESBD shows similar safety level,and is safer than EPBD.So ESBD is a safe and effective method to remove CBDs with PAD.

Objective To study the clincial effectiveness and safety of endoscopic sphincteropapillotomy combined with balloon dilation for decompensated cirrhosis accompanied with choledocholithiasis.Methods Data of 79 cases of decompensated cirrhosis patients with choledocholithiasis who underwent limited endoscopic sphincterotomy plus endoscopic papillary balloon dilation (ESBD,the ESBD group) and 42 cases who underwent endoscopic papillary balloon dilation(EPBD,the EPBD group)were retrospectively analysed and compared for complete stone clearance rate,one-time stone clearance rate and complications.Results The rate of complete stone clearance and one-time stone clearance were 94.9％ (75/79) and 77.2％ (61/79)in ESBD group,and those were 88.1％ (37/42) and 59.5％ (25/42) in EPBD group respectively.The rate of complete stone clearance was a little higher in ESBD group than that in EPBD group.In ESBD group,ERCP-related bleeding occurred in 3 patients (3.8％),post-ERCP hyperamylasemia in 3 (3.8％)and post-ERCP pancreatitis in 2 (2.5％) ; while in EPBD group,post-ERCP hyperamylasemia occurred in 8 patients(19.0％),post-ERCP pancreatitis in 6(14.3％) and ERCP-related bleeding did not occur.There were no significant difference in ERCP-related bleeding between ESBD group and EPBD group (P =0.551).However,the rates of post-ERCP pancreatitis and hyperamylasemia in ESBD group were significantly lower than those in EPBD group(P ＜ 0.05).Conclusion ESBD is a safe and effective procedure for choledocholithiasis accompanied by decompensated cirrhosis,with several advantages over EPBD in terms of higher one-time stone clearance rate,reduced risk of post-ERCP pancreatitis and hyperamylasemia,and without noticeable increase in the risk of bleeding related to ERCP.

ObjectiveTo identify the curative effect of minimally invasive plate osteosynthesis (MIPO) and humeral head replacement in treatment of elderly patients with Neer four-part fractures.MethodsUsing the deltoid pectoral approach,28 patients with fresh Neer four-part fractures were treated by the locking plate combined with MIPO (Group A) and 27 by the humeral head replacement (Group B).Neer score,Constant-Murley score and simple shoulder test (SST) questionnaire were adopted for assessing the treatment outcome.ResultsGroup A was followed up for mean 32.1 months,which showed screws protruding into the joint space in two patients,tuberosity upward and backward displacement in one and femoral head ischemic necrosis in one.The mean visual analog scale (VAS) score,the mean Neer score and the mean Constant-Murley score were 2.2 points,88.6 points and 86.5 points respectively.There were average 9.0 answers for “yes” in the SST questionnaire.Group B were followed up for mean 34.6 months,which showed shoulder dislocation or subluxation in four patients,tuberosity displacement or excessive reduction in eight.The mean VAS score,the mean Neer score and the mean Constant-Murley score were 2.4 points,78.9 points and 77.3 points respectively.The mean number of answer for “yes” in SST questionnaire was 8.0 questions.There showed no statistical difference in VAS score between the two groups.While statistical difference was found in complications,Neer score,Constant-Murley score and SST score between two groups,with Group A superior to Group B.Conclusions For most elderly patients with Neer four-part fractures,MIPO has satisfactory results under strict control of surgical indications and technical tips.The humeral head replacement surgery still has many unresolved problems and needs careful consideration.