MLL4 Regulates the Progression of Non–Small-Cell Lung Cancer by Regulating the PI3K/AKT/SOX2 Axis / Journal of the Korean Cancer Association, 대한암학회지

Purpose@#Mixed-lineage leukemia protein 4 (MLL4/KMT2D) is a histone methyltransferase, and its mutation has been reported to be associated with a poor prognosis in many cancers, including lung cancer. We investigated the function of MLL4 in lung carcinogenesis. @*Materials and Methods@#RNA sequencing (RNA-seq) in A549 cells transfected with control siRNA or MLL4 siRNA was performed. Also, we used EdU incorporation assay, colony formation assays, growth curve analysis, transwell invasion assays, immunohistochemical staining, and in vivo bioluminescence assay to investigate the function of MLL4 in lung carcinogenesis. @*Results@#We found that MLL4 expression was downregulated in non–small cell lung cancer (NSCLC) tissues compared to adjacent normal tissues and tended to decrease with disease stage progression. We analyzed the transcriptomes in control and MLL4- deficient cells using high-throughput RNA deep sequencing (RNA-seq) and identified a cohort of target genes, such as SOX2, ATF1, FOXP4, PIK3IP1, SIRT4, TENT5B, and LFNG, some of which are related to proliferation and metastasis. Our results showed that low expression of MLL4 promotes NSCLC cell proliferation and metastasis and is required for the maintenance of NSCLC stem cell properties. @*Conclusion@#Our findings identify an important role of MLL4 in lung carcinogenesis through transcriptional regulation of PIK3IP1, affecting the PI3K/AKT/SOX2 axis, and suggest that MLL4 could be a potential prognostic indicator and target for NSCLC therapy.

Advances in Chemical Constituents and Pharmacological Action of Tinosporae Radix / 中国实验方剂学杂志

Tinosporae Radix， as a traditional Chinese medicinal herb， is the dried root tuber of Tinospora sagittata or T. capillipes. It was first recorded in the Compendium of Materia Medica Supplement in the Qing Dynasty and included in the previous edition of the Chinese Pharmacopoeia. Tinosporae Radix is excavated in autumn and winter and used after removing fibrous roots， washing， and drying. It is indicated for sore throat， carbuncle boils poison， waist and abdominal pain， and various heat syndromes and is commonly used to treat chronic inflammation. Its efficacy is significantly known as “broad-spectrum antibiotics in Zhuang medicine”. Tinosporae Radix is a traditional Chinese medicinal herb often taken by Zhuang and Yao nationalities in Guangxi province and has a wide range of application and development values and research significance. Modern studies have shown that Tinosporae Radix contains diterpenoids， alkaloids， sterols， anthraquinones， glycosides， fatty acids， volatile oils， and other compounds， which have many pharmacological activities such as anti-inflammatory and analgesic， antibacterial and antibacterial， antioxidant， anti-diabetic， and anti-tumor and anti-cancer effects， and it has achieved good efficacy in inhibiting inflammation and treating sore throat and other diseases. In recent years， there have been many research reports on the status， chemical constituents， pharmacological action， clinical application， and quality evaluation of Tinosporae Radix resources， but there is no systematic review and introduction at present. By consulting the literature and combining it with modern research， this paper systematically summarizes and collates Tinosporae Radix resources to provide guidance for the comprehensive development and utilization of Tinosporae Radix resources and subsequent in-depth study.

GPRC5B protects osteoarthritis by regulation of autophagy signaling

Osteoarthritis (OA) is one of the most common chronic diseases in the world. However, current treatment modalities mainly relieve pain and inhibit cartilage degradation, but do not promote cartilage regeneration. In this study, we show that G protein-coupled receptor class C group 5 member B (GPRC5B), an orphan G-protein-couple receptor, not only inhibits cartilage degradation, but also increases cartilage regeneration and thereby is protective against OA. We observed that Gprc5b deficient chondrocytes had an upregulation of cartilage catabolic gene expression, along with downregulation of anabolic genes in vitro. Furthermore, mice deficient in Gprc5b displayed a more severe OA phenotype in the destabilization of the medial meniscus (DMM) induced OA mouse model, with upregulation of cartilage catabolic factors and downregulation of anabolic factors, consistent with our in vitro findings. Overexpression of Gprc5b by lentiviral vectors alleviated the cartilage degeneration in DMM-induced OA mouse model by inhibiting cartilage degradation and promoting regeneration. We also assessed the molecular mechanisms downstream of Gprc5b that may mediate these observed effects and identify the role of protein kinase B (AKT)-mammalian target of rapamycin (mTOR)-autophagy signaling pathway. Thus, we demonstrate an integral role of GPRC5B in OA pathogenesis, and activation of GPRC5B has the potential in preventing the progression of OA.

Clinical characteristics and prognosis of multiple primary colorectal carcinoma / 肿瘤研究与临床

Objective:To explore the clinical characteristics and prognosis of multiple primary colorectal carcinoma.Methods:The clinical data of 42 cases of colorectal cancer admitted to the First Affiliated Hospital of Nanjing Medical University between January 2013 and December 2018 were retrospectively analyzed. The clinicopathological features, diagnosis, treatment and prognosis were summarized.Results:There were 42 patients with multiple primary colorectal carcinoma, accounting for 1.20% (42/3 499) of all colorectal carcinoma patients in the same period. The main pathological type was adenocarcinoma. Among them, 32 cases were synchronous multiple primary carcinoma. And the age ranged 38-86 years old, and the median age was 66 years old. A total of 73 colorectal cancer lesions were detected, mostly located in the proximal colon, sigmoid colon and rectum. A total of 527 lymph nodes were detected, and the positive rate was 1.9% (10). Patients with positive lymph nodes accounted for 37.5% (12/32), including 27 cases of multiple primary carcinoma, 3 cases of triple primary carcinoma, 2 cases of five primary carcinoma. The 1-year and 3-year overall survival rates were 83.75% and 74.38%, respectively. There were 10 cases of metachronous multiple primary carcinoma. Patients were aged 33-86 years old. The first cancer was mostly located in the rectum and sigmoid colon, and the second cancer was mostly located in the ascending colon area. A total of 276 lymph nodes were detected, and the positive rate was 12.3% (34). The 1-year and 3-year overall survival rates were 100.00% and 66.67%, respectively.Conclusions:Multiple primary colorectal cancer is not rare clinically and its distribution shows a certain regularity. More attention should be paid to improve the early diagnosis rate. Early operation is needed to improve the survival rate of patients.

Application value of superior mesenteric artery-oriented complete mesocolic excision in the treatment of right colon cancer / 中华消化外科杂志

Objective To investigate the application value of superior mesenteric artery (SMA)-oriented complete mesocolic excision (CME) in the treatment of right colon cancer.Methods The retrospective cohort study was conducted.The clinicopathological data of 955 patients with right colon cancer who were admitted to the First Affiliated Hospital of Nanjing Medical University from January 2013 to June 2018 were collected.There were 514 males and 441 females,aged from 18 to 96 years,with a median age of 65 years.Of the 955 patients,377 undergoing SMA-oriented CME of right colon with the lymph node dissection along the left boundary of SMA were allocated into SMA-oriented group,and 578 undergoing superior mesenteric vein (SMV)-oriented CME of right colon with the lymph node dissection along the left boundary of SMV were allocated into SMV-oriented group.Observation indicators:(1) intraoperative and postoperative conditions;(2) postoperative complications;(3) postoperative pathological examinations;(4) follow-up and survival situations.Follow-up was performed by telephone interview and outpatient examination once every 3-6 months within 2 years after surgery and once a year after 2 years up to January 2019,using tumor recurrence and metastasis or death as the end point.Follow-up included physical examination and tumor marker test,including carcino embryonic antigen,CA19-9,chest and abdomen CT examination and enteroscopy.Measurement data with skewed distribution were described as M (P25,P75),and comparison between groups was done using the Mann-Whitney U test.Count data were described as absolute numbers or percentages,and comparison between groups was analyzed using the chi square test.Ordinal data were analyzed using the rank sum test.Kaplan-Meier method was used to calculate survival time and rate,and draw survival curve.Log-rank test was used for survival analysis.Patients with loss to follow-up were involved in survival analysis as censored data.Results (1) Intraoperative and postoperative conditions:the operation time,volume of intraoperative blood loss,duration of postoperative hospital stay were 100 minutes (90 minutes,110 minutes),50.0 mL (50.0 mL,70.0 mL),8 days (8 days,10 days) in the SMA-oriented group,and 110 minutes (90 minutes,135 minutes),50.0 mL (50.0 mL,122.5 mL),10 days (8 days,12 days) in the SMV-oriented group,showing significant differences between the two groups (Z=-5.400,-5.799,-7.461,P＜0.05).After the exclusion of 47 patients unsuitable for defecation analysis because of postoperative complications,365 in the SMA-oriented group and 543 in the SMV-oriented group were analyzed.The time to first defecation,the maximum number and the median number of daily defecation postoperatively were 5 days (3 days,5 days),2.0 (1.0,2.5),1.0 (1.0,1.0) in the SMA-oriented group,which showed no significant difference from 4 days (3 days,5 days),2.0 (1.0,3.0),1.0 (1.0,1.0) in the SMV-oriented group (Z=-1.622,-1.541,-1.024,P＞ 0.05).(2) Postoperative complications:cases with postoperative complications,cases with incisional liquefaction or infection,cases with anastomostic leakage,cases with delayed gastric emptying,cases with intra-abdominal bleeding,cases with complete or incomplete ileus,cases with anastomostic bleeding,cases with intra-abdominal infection,cases with disruption of wound,the number of death were 55,10,3,3,2,2,1,1,1,1 in the SMA-oriented group,which showed no significant difference from 83,30,13,4,3,8,3,6,2,3 in the SMV-oriented group,respectively (x2 =0.045,3.662,2.926,0.034,0.001,1.604,0.352,1.873,0.048,0.352,P＞0.05).There were 32 of 377 patients in the SMA-oriented group and 14 of 578 in the SMV-oriented group with chylous leakage,showing a significant difference between the two groups (x2 =18.312,P＜ 0.05).Patients with chylous leakage were improved after conservative treatment,without reoperation.Patients with other complications were improved after anti-infection,fluid infusion,and reoperation.Four of 955 patients died after surgery.(3) Postoperative pathological examinations:patients with stage Ⅰ,stage Ⅱ,and stage Ⅲ (pathological TNM staging),patients with high-differentiation,mid-differentiation,and low-differentiation (tumor differentiation degree),length of intestine specimen,number of positive lymph nodes,maximum tumor diameter,patients with cancer nodules,patients with vascular invasion,patients with perineural invasion were 57,174,146,30,174,173,23 cm (21 cm,26 cm),0 (0,2),5.0 cm (3.0 cm,6.0 cm),37,81,53 in the SMA-oriented group,which showed no difference from 66,280,232,33,303,242,23 cm (21 cm,25 cm),0 (0,2),5.0 cm (3.5 cm,6.0 cm),80,108,82 in the SMV-oriented group (Z=-1.020,-0.216,-0.243,-0.220,-0.814,x2=3.441,1.127,0.003,P＞0.05).The number of harvested lymph nodes was 22.0 (17.0,27.0) and 18.0 (15.0,22.0) in the SMA-oriented group and SMV-oriented group,respectively,with a significant difference between the two groups (Z=-7.800,P＜0.05).There were 202 patients extracted for further analysis.The number of harvested lymph nodes and harvested central lymph nodes was 25.0 (20.0,31.3),5.0 (3.0,8.0) of 166 patients in the SMA-oriented group,and 21.5 (18.0,28.8),1.5 (0,4.5) of 36 patients in the SMV-oriented group,respectively,showing significant differences between the two groups (Z =-1.995,-4.309,P＜0.05).(4) Follow-up and survival situations:840 of 955 patients including 346 in the SMA-oriented group and 494 in the SMV-oriented group were followed up for 1.0-73.2 months,with a median time of 31.5 months.SMA-oriented group had a higher 5-year overall survival rate than SMV-oriented group (91.8％ vs.84.9％,x2 =4.384,P＜0.05),but had no significant difference in the 5-year tumor-free survival rate compared with the SMV-oriented group (84.4％ vs.78.2％,x2=2.158,P＞0.05).Conclusion Compared with SMV-oriented CME of right colon,SMA-oriented CME of right colon is safe and feasible,with larger number of harvested lymph nodes,which can achieve complete lymph node dissection.

Clinical effects of laparoscopic left colic artery-preserving total mesorectal excison for middle-low rectal cancer / 中华消化外科杂志

Objective To investigate the clinical effects of laparoscopic left colic artery (LCA) preserving total mesorectal excision (TME) for middle-low rectal cancer.Methods The retrospective cohort study was conducted.The clinicopathological data of 339 patients with middle-low rectal cancer who underwent laparoscopic TME in the First Affiliated Hospital of Nanjing Medical University from July 2016 to July 2017 were collected.There were 216 males and 123 females,aged from 35 to 89 years,with an average age of 62 years.No lymph node enlargement was detected at the root of inferior mesenteric artery as evaluated by preoperative examination.Of the 339 patients,173 undergoing laparoscopic TME with preservation of LCA and 166 without preservation of LCA were allocated into observation group and control group,respectively.Observation indicators:(1) surgical situations;(2) postoperative situations;(3) follow-up.Follow-up using outpatient examination and telephone interview was performed to detect complications and survival of patients after hospital discharge up to August 2018.Measurement data with normal distribution were represented as Mean±SD,and comparison between groups was analyzed using the t test.Measurement data with skewed distribution were described as M (range),and comparison between groups was analyzed using the Mann-Whitney U test.Count data were represented as absolute number or percentage,and comparison between groups was analyzed using the chi-square test or Fisher exact propability.Comparisons of ordinal data were analyzed by Mann-Whitney U test.Results (1) Surgical situations:339 patients underwent laparoscopic TME successfully,without conversion to open surgery or death.The operation time,volume of intraoperative blood loss and number of lymph nodes dissected at the root of inferior mesenteric artery were (77± 15)minutes,(32±22)mL and 3.5± 1.3 in the observation group,(74± 16)minutes,(30±21) mL and 3.6± 1.3 in the control group,respectively,showing no statistically significant difference between the two groups (t =1.730,0.790,-0.378,P＞0.05).There were 21 and 37 patients receiving preventive stoma in the observation group and control group,with a statistically significant difference between the two groups (x2 =6.154,P＜0.05).(2) Postoperative situations:no lymphatic metastasis at the root of inferior mesenteric artery was detected in the observation group or control group.The incidence of anastomotic leakage was 8.55％ (13/152) and 16.28％(21/129) of patients without preventive stoma in the observation group and control group,with a statistically significant difference between the two groups (x2=3.917,P＜0.05).The overall incidence of anastomotic leakage was 7.51％ (13/173) and 12.65％ (21/166),with no statistically significant difference between the two groups (x2 =2.477,P＞0.05).Cases with anastomotic hemorrhage and time to first anal sufflation were 18 and (2.2±0.9) days in the observation group,10 and (2.4±1.0) days in the control group,respectively,showing no statistically significant difference between the two groups (x2 =2.145,t =-1.370,P＞0.05).(3) Follow-up:339 patients were followed up for 13-22 months,with a median time of 14 months.During the follow-up,2 patients had postoperative intestinal obstruction,19 had tumor recurrence or metastasis,and 4 died in the observation group.In the control group,1 patient had postoperative intestinal obstruction,17 had tumor recurrence or metastasis,and 3 died.The other patients in the two groups survived well.Conclusion Laparoscopic TME with preservation of LCA can reduce incidence of anastomotic leakage in patients with middle-low rectal cancer who don't undergo preventive stoma.

Risk factors analysis of anastomotic leakage after low anterior resection of rectal cancer / 国际外科学杂志

Objective To analyze the risk factors for anastomotic leakage after low anterior resection(LAR) for rectal cancer.Methods The retrospective study of 1 336 patients with rectal cancer who underwent LAR from Jan.2013 to Dec.2017 in the Department of Colorectal Surgery,First Affiliated Hospital of Nanjing Medical University was conducted.There were 838 male and 498 female patients.The median age was 62 (53,70) years.Observation indicators:①intraoperative conditions,including the performance of preventive enterostomy,the preservation of left colic artery,the placement of transanal decompression tube;②postoperative conditions,for example,postoperative clinical symptoms and physical examination,postoperative hospital stay.Anastomotic leakage patients were graded according to the grading system of anastomotic leakage following anterior resection of the rectum proposed by the International Study Group of Rectal Cancer in 2010.Measurement data with non-normal distribution were described as median,and comparison between groups was done using Mann-Whitney U test.Ranked data were compared with rank sum test.Comparison of count data between groups were analyzed using the Chi-square test or Fisher exact test.Potential risk factors for anastomotic leakage were further analyzed with multiple logistic regression.Results Anastomotic leakage was noted in 138 (10.4％) of 1 336 patients with 93 patients (7.0％) of Grade B and 46 patients (3.4％) of Grade C respectively.The occurrence of Grade C anastomotic leakage was on the 3th day postoperatively,which was earlier than Grade B anastomotic leakage [5 (3,7) d,Z =2.746,P =0.006].There were significant differences in sex,BMI,placement of transanal decompression tube and tumor location between anastomotic leakage cases and non-anastomotic leakage cases by Chi-squared test (x2 =10.897,9.033,6.343,6.34,Z =-2.380,P ＜0.05).On multiple logistic regression analysis,male sex (OR =1.840,95 ％ CI:1.210-2.799,P =0.004),BMI ≥ 24 kg/m2 (OR =1.573,95 ％ CI:1.088-2.274,P =0.016),placement of transanal decompression tube(OR =2.418,95％ CI:1.196-4.888,P =0.014),middle rectal cancer (OR =1.536,95％ CI:O.836-2.822,P =0.167),low rectal cancer(OR =1.989,95％ CI:1.068-2.822,P =0.03),ultralow rectal cancer (OR =2.908,95％ CI:1.289-6.560,P =0.01) were independent risk factors of anastomotic leakage.Conclusion Male sex,high BMI,placement of transanal decompression tube and low rectal tumor were independent risk factors of anastomotic leakage for patients receiving LAR.

Neuroprotective effect of rapamycin against Parkinson's disease in mice / 浙江大学学报·医学版

OBJECTIVE@#To investigate the effect of rapamycin on Parkinson's disease (PD) and its underlying mechanism in mice.@*METHODS@#Sixty SPF adult male C57BL/6 mice were randomly divided into control group, model group and treatment group. 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine(MPTP) was used to induce Parkinson's disease in model group and treatment group. All mice were trained to cross the runway and were subjected to computer-assisted CatWalk. The numbers of tyrosine hydroxylase positive (TH) neurons in the substantia nigra (SN) were assessed by unbiased stereology using the optical fractionator method; protein expression was detected by Western blot analysis; and glutathione peroxidase (GSH-Px), malondialdehyde (MDA) and superoxide dismutase (SOD) were detected by spectrophotometry.@*RESULTS@#In the model group, a decrease in stride rate and an increase in variation of stance and swing were noted by CatWalk system (<0.05 or <0.01); the numbers of TH neurons decreased (<0.01); expression of p-Akt, p-S6K, p-S6 and p-ULK increased (all <0.01); LC3-Ⅱ/Ⅰ ratio decreased (<0.01); MDA level was elevated while the levels of SOD and GSH-PX were reduced (all <0.01). Compared with the model group, after treated with rapamycin, the abnormal behavior including the stride length, variation of stance and swing and step patterns induced by MPTP were all improved (<0.05 or <0.01); the numbers of TH neurons increased (<0.05); the expression of p-Akt, p-S6K, p-S6 and p-ULK was suppressed (all <0.01); the LC3-Ⅱ/Ⅰ ratio was upregulated (<0.05); MDA level decreased while the levels of GSH-Px and SOD increased (all <0.01).@*CONCLUSIONS@#Rapamycin inhibits the activation of mTOR pathway, which contributes to protect against the loss of dopaminergic neurons and provide behavioral improvements in mice with Parkinson's disease. These results are partially related to the ability of rapamycin in inducing autophagy and reducing oxidative stress.

Clinical efficacy of superior mesenteric artery-oriented laparoscopic complete mesocolic excision in the treatment of right colonic cancer / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To investigate the clinical efficacy of superior mesenteric artery (SMA)-oriented laparoscopic complete mesocolic excision (CME) with a medial-to-lateral approach in the treatment of right colonic cancer.</p><p><b>METHODS</b>A total of 166 patients with right colonic cancer who were admitted to The First Affiliated Hospital of Nanjing Medical University from January 2015 to December 2016 underwent SMA-oriented laparoscopic CME by the same surgical team. Clinical data of these patients were retrospectively analyzed.</p><p><b>RESULTS</b>All the 166 cases were enrolled in this study, including 96 male and 70 female patients. Right colon adenocarcinoma was diagnosed by preoperative colonoscopy, enhanced computerized tomography(CT) and pathological examination, and the tumor was resectable. There were no emergency surgery patients for bowel obstruction or perforation, no patients with multiple primary cancers, and no stage IIII( patients with liver or lung metastasis. All the patients underwent SMA-oriented laparoscopic CME successfully, without conversion to laparotomy. No ureteral injury, duodenal injury, and superior mesenteric vein injury occurred during the operation. R0 resections were performed in all the cases. The average operation time was (102.4±28.3) min, blood loss was (67.2±26.3) ml, number of lymph nodes retrieved was 20.5±7.6, postoperative anal exhaust time was (2.1±1.1) d, postoperative hospital stay was (7.9±1.4) d. There was no perioperative death or anastomotic leakage. Two patients with small bowel obstruction were readmitted within 30 days after operation. The postoperative complications occurred in 15.1%(25/166) of the patients, including wound infection (2 cases), chylous fistula (20 cases), anastomotic bleeding (1 case), small bowel obstruction (2 cases). A total of 161 patients (97.0%) were followed up for 1 to 24 months, and 10 patients had distant metastasis during the follow-up period. The 2-year overall survival (OS) rate was 96.4%.</p><p><b>CONCLUSION</b>Superior mesenteric artery-oriented laparoscopic complete mesocolic excision with a medial-to-lateral approach for right colonic cancer has good short-term efficacy.</p>