1.A nationwide multicenter prospective study on the perioperative impact of closure of mesen-teric fissure in laparoscopic right hemicolectomy
Gang LIU ; Weimin XU ; Da LI ; Lei QIAO ; Jieqing YUAN ; Dewei ZHANG ; Yan LIU ; Shuai GUO ; Xu ZHANG ; Wenzhi LIU ; Yingfei WANG ; Hang LU ; Xiaowei ZHANG ; Xin CHEN ; Zhaohui XU ; Xingyang LUO ; Ge LIU ; Cheng ZHANG ; Jianping ZHOU
Chinese Journal of Digestive Surgery 2024;23(6):812-818
		                        		
		                        			
		                        			Objective:To investigate the perioperative impact of closure of mesenteric fissure in laparoscopic right hemicolectomy.Methods:The prospective randomized controlled trial was conducted. The clinical data of 320 patients who underwent laparoscopic right hemicolectomy in 11 medical centers, including The First Affiliated Hospital of China Medical University et al, from November 2022 to August 2023 were selected. Based on block randomization, patients were alloca-ted into the mesenteric fissure non-closure group and the mesenteric fissure closure group. Observa-tion indicators: (1) grouping of the enrolled patients; (2) intraoperative conditions; (3) postopera-tive conditions. Measurement data with skewed distribution were represented as M( Q1, Q3) and com-parison between groups was conducted using the Mann-Whitney U test. Count data were represen-ted as absolute numbers or percentages, and comparison between groups was conducted using the chi-square test or Fisher's exact probability. Comparison of ordinal data was conducted using the rank sum test. Comparison of visual analog scores was analyzed using generalized estimating equations. Results:(1) Grouping of the enrolled patients. A total of 320 patients with colon cancer were screened for eligibility, including 156 males and 164 females, aged 68(59,73)years. All the 320 patients were allocated into the mesenteric fissure non-closure group with 164 cases and the mesenteric fissure closure group with 156 cases. There was no significant difference in the age, body mass index, American Society of Anesthesiologist score, maximum tumor diameter, anastomosis location, anastomosis method, surgical approach, range of lymph node dissection, tumor staging between the two groups ( P>0.05) and there was a significant difference in the sex between them ( P<0.05). (2) Intraoperative conditions. There was no significant difference between the mesenteric fissure closure group and the mesenteric fissure non-closure group in the volume of intraoperative blood loss, operation time, conversion to laparotomy, intraoperative complication ( P>0.05). Three patients in the mesenteric fissure non-closure group were converted to laparotomy. One patient in the mesenteric fissure closure group was converted to laparotomy, and 2 cases with intraoperative complication were mesenteric hematoma. (3) Postoperative conditions. There was no significant difference between the mesenteric fissure non-closure group and the mesenteric fissure closure group in the overall postoperative complications ( χ2=0.28, P>0.05). There was no significant difference in the occurrence of postoperative intestinal obstruction, abdominal distension, ascites, pleural effusion, gastric paralysis, anastomotic bleeding, anastomotic leakage, or surgical wound infection between the two groups ( P>0.05). There was no significant difference between the two groups in the reoperation, postoperative gastric tube replacement. There was no significant differ-ence in time to postoperative first flatus, time to postoperative initial liquid food intake, time to post-operative resumption of bowel movements, duration of postoperative hospital stay, total hospital expenses between the two groups ( Z=-0.01, 0.43, 1.04, -0.54, -0.36, P>0.05). One patient in the mesenteric fissure non-closure group received reoperation. No perioperative internal hernia or death occurred in either group. The visual analog score decreased with time in both groups. There was no significant difference in the visual analog score between the mesenteric fissure closure group and the mesenteric fissure non-closure group [ β=-0.20(-0.53,0.13), P>0.05]. Conclusion:Compared with closure of mesenteric fissure, non-closure of mesenteric fissure during laparoscopic right hemi-colectomy dose not increase perioperative complications or postoperative management risk.
		                        		
		                        		
		                        		
		                        	
2.Impact of unidentified injectable fillers on nasal dorsal soft tissue and rhinoplasty
Zhengchao LI ; Chen ZHANG ; Weiyan CHEN ; Bo HU ; Jieqing WANG ; Xiaowei SU
Chinese Journal of Medical Aesthetics and Cosmetology 2024;30(3):256-261
		                        		
		                        			
		                        			Objective:To explore the impact of unidentified injectable fillers on the soft tissue of nasal dorsum and rhinoplasty.Methods:The Plastic Surgery Information System of Xinhua Hospital Affiliated with Dalian University was utilized to conduct an analysis of 62 rhinoplasty patients between 2018 and 2019. Specifically, this included 28 patients with an unidentified history of injectable filler rhinoplasty, encompassing 1 male and 27 females with ages ranging from 19 to 53 years and a mean age of 28.8 years. Additionally, 34 patients underwent primary rhinoplasty, all of whom were female with ages ranging from 19 to 46 years and a mean age of 26.8 years. This study examined the effects of unidentified injectable fillers on the soft tissue of the nasal dorsum by analyzing the excised nasal dorsum under a microscope. Subsequently, statistical methods were performed to assess differences in gender, age, preoperative tip protrusion/nose length, postoperative tip protrusion/nose length, dorsal augmentation modality, and satisfaction, and to investigate the effect of unidentified injectables on the rate of dissatisfaction after rhinoplasty.Results:The histopathological analysis of unidentified injectable fillers removed from the nasal dorsum revealed the presence of mainly gel and granular fillers. The gel fillers, characterized by its pink jelly-like texture, contained unidentified injectable fillers, colorless glue, and were observed to flow out upon cutting. The granular filler, on the other hand, appeared as tough, irregularly shaped tissue similar to caviar. Additionally, evidence of muscle tissue in 5 pathologic sections indicated that the unidentified injectable fillers were injected into or near the dorsal nasal muscles, leading to varying degrees of muscle injuries upon excision. A comparison of 28 rhinoplasty patients with unidentified injectable fillers for nasal dorsal augmentation and 34 patients with primary rhinoplasty showed that 11 females in the former group and 4 females in the latter group were dissatisfied with the results. Statistical analysis demonstrated no significant differences between the two groups in terms of gender ( P=0.452), age ( P=0.219), preoperative tip projection/nasal length ( P=0.681), postoperative tip projection/nasal length ( P=0.105), and nasal dorsum augmentation methods ( P=0.413). However, the initial rhinoplasty group had a lower dissatisfactory rate (4 cases, 11.76%) and the unidentified injectables group had a higher dissatisfactory rate (11 cases, 39.29%), which was statistically significant between the two groups (χ 2=6.341, P=0.012). Conclusions:The presence of unidentified nasal injectable fillers can significantly decrease postoperative satisfactory rates, increase the incidence of dissatisfaction, and have adverse effects on the soft tissues of the nasal dorsum and the overall outcome of the rhinoplasty procedure.
		                        		
		                        		
		                        		
		                        	
3.First-line bevacizumab plus chemotherapy in Chinese patients with stage III/IV epithelial ovarian cancer, fallopian tube cancer or primary peritoneal cancer: a phase III randomized controlled trial
Xiaohua WU ; Jihong LIU ; Ruifang AN ; Rutie YIN ; Yu ZHANG ; Huaijun ZHOU ; Aiqin HE ; Li WANG ; Jieqing ZHANG ; Ziling LIU ; Wei DUAN ; Jianqing ZHU ; Ge LOU ; Guilin CHEN ; Ying CHENG ; Fengxia XUE ; Sonja NICK ; Haiyan WANG ; Donghang LI
Journal of Gynecologic Oncology 2024;35(5):e99-
		                        		
		                        			 Objective:
		                        			First-line bevacizumab plus carboplatin and paclitaxel (CP) is approved for stage III/IV ovarian cancer treatment following initial surgical resection, based on global phase III GOG-0218 and ICON7 trials. This study evaluated the efficacy and safety of bevacizumab + CP as first-line ovarian cancer therapy in Chinese patients. 
		                        		
		                        			Methods:
		                        			Patients with newly diagnosed, International Federation of Gynecology and Obstetrics (FIGO) stage III/IV epithelial ovarian, fallopian tube, or primary peritoneal cancer post-primary surgery were randomized 1:1 to receive 6 cycles of CP with bevacizumab/ placebo, followed by bevacizumab/placebo maintenance until unacceptable toxicity or disease progression. Primary endpoint was investigator-assessed progression-free survival (PFS). Stratification factors were FIGO stage and debulking status (stage III optimally debulked vs stage III suboptimally debulked vs stage IV) and Eastern Cooperative Oncology Group performance status (0 vs 1 or 2). 
		                        		
		                        			Results:
		                        			Of randomized patients, 51 received bevacizumab + CP and 49 received placebo + CP. Median PFS was 22.6 months with bevacizumab + CP (95% confidence interval [CI]=18.6, not estimable) and 12.3 months (95% CI=9.5, 15.0) with placebo + CP (stratified hazard ratio=0.30; 95% CI=0.17, 0.53). Treatment-related grade 3/4 adverse events occurred in 46 of 49 (94%) patients receiving bevacizumab + CP, and 34 of 50 (68%) receiving placebo + CP. 
		                        		
		                        			Conclusion
		                        			Bevacizumab + CP showed clinically meaningful improvement in PFS vs placebo + CP, consistent with GOG-0218 results. Safety data were aligned with the known bevacizumab safety profile. These results support first-line bevacizumab + CP therapy in Chinese patients with ovarian cancer. 
		                        		
		                        		
		                        		
		                        	
4.First-line bevacizumab plus chemotherapy in Chinese patients with stage III/IV epithelial ovarian cancer, fallopian tube cancer or primary peritoneal cancer: a phase III randomized controlled trial
Xiaohua WU ; Jihong LIU ; Ruifang AN ; Rutie YIN ; Yu ZHANG ; Huaijun ZHOU ; Aiqin HE ; Li WANG ; Jieqing ZHANG ; Ziling LIU ; Wei DUAN ; Jianqing ZHU ; Ge LOU ; Guilin CHEN ; Ying CHENG ; Fengxia XUE ; Sonja NICK ; Haiyan WANG ; Donghang LI
Journal of Gynecologic Oncology 2024;35(5):e99-
		                        		
		                        			 Objective:
		                        			First-line bevacizumab plus carboplatin and paclitaxel (CP) is approved for stage III/IV ovarian cancer treatment following initial surgical resection, based on global phase III GOG-0218 and ICON7 trials. This study evaluated the efficacy and safety of bevacizumab + CP as first-line ovarian cancer therapy in Chinese patients. 
		                        		
		                        			Methods:
		                        			Patients with newly diagnosed, International Federation of Gynecology and Obstetrics (FIGO) stage III/IV epithelial ovarian, fallopian tube, or primary peritoneal cancer post-primary surgery were randomized 1:1 to receive 6 cycles of CP with bevacizumab/ placebo, followed by bevacizumab/placebo maintenance until unacceptable toxicity or disease progression. Primary endpoint was investigator-assessed progression-free survival (PFS). Stratification factors were FIGO stage and debulking status (stage III optimally debulked vs stage III suboptimally debulked vs stage IV) and Eastern Cooperative Oncology Group performance status (0 vs 1 or 2). 
		                        		
		                        			Results:
		                        			Of randomized patients, 51 received bevacizumab + CP and 49 received placebo + CP. Median PFS was 22.6 months with bevacizumab + CP (95% confidence interval [CI]=18.6, not estimable) and 12.3 months (95% CI=9.5, 15.0) with placebo + CP (stratified hazard ratio=0.30; 95% CI=0.17, 0.53). Treatment-related grade 3/4 adverse events occurred in 46 of 49 (94%) patients receiving bevacizumab + CP, and 34 of 50 (68%) receiving placebo + CP. 
		                        		
		                        			Conclusion
		                        			Bevacizumab + CP showed clinically meaningful improvement in PFS vs placebo + CP, consistent with GOG-0218 results. Safety data were aligned with the known bevacizumab safety profile. These results support first-line bevacizumab + CP therapy in Chinese patients with ovarian cancer. 
		                        		
		                        		
		                        		
		                        	
5.First-line bevacizumab plus chemotherapy in Chinese patients with stage III/IV epithelial ovarian cancer, fallopian tube cancer or primary peritoneal cancer: a phase III randomized controlled trial
Xiaohua WU ; Jihong LIU ; Ruifang AN ; Rutie YIN ; Yu ZHANG ; Huaijun ZHOU ; Aiqin HE ; Li WANG ; Jieqing ZHANG ; Ziling LIU ; Wei DUAN ; Jianqing ZHU ; Ge LOU ; Guilin CHEN ; Ying CHENG ; Fengxia XUE ; Sonja NICK ; Haiyan WANG ; Donghang LI
Journal of Gynecologic Oncology 2024;35(5):e99-
		                        		
		                        			 Objective:
		                        			First-line bevacizumab plus carboplatin and paclitaxel (CP) is approved for stage III/IV ovarian cancer treatment following initial surgical resection, based on global phase III GOG-0218 and ICON7 trials. This study evaluated the efficacy and safety of bevacizumab + CP as first-line ovarian cancer therapy in Chinese patients. 
		                        		
		                        			Methods:
		                        			Patients with newly diagnosed, International Federation of Gynecology and Obstetrics (FIGO) stage III/IV epithelial ovarian, fallopian tube, or primary peritoneal cancer post-primary surgery were randomized 1:1 to receive 6 cycles of CP with bevacizumab/ placebo, followed by bevacizumab/placebo maintenance until unacceptable toxicity or disease progression. Primary endpoint was investigator-assessed progression-free survival (PFS). Stratification factors were FIGO stage and debulking status (stage III optimally debulked vs stage III suboptimally debulked vs stage IV) and Eastern Cooperative Oncology Group performance status (0 vs 1 or 2). 
		                        		
		                        			Results:
		                        			Of randomized patients, 51 received bevacizumab + CP and 49 received placebo + CP. Median PFS was 22.6 months with bevacizumab + CP (95% confidence interval [CI]=18.6, not estimable) and 12.3 months (95% CI=9.5, 15.0) with placebo + CP (stratified hazard ratio=0.30; 95% CI=0.17, 0.53). Treatment-related grade 3/4 adverse events occurred in 46 of 49 (94%) patients receiving bevacizumab + CP, and 34 of 50 (68%) receiving placebo + CP. 
		                        		
		                        			Conclusion
		                        			Bevacizumab + CP showed clinically meaningful improvement in PFS vs placebo + CP, consistent with GOG-0218 results. Safety data were aligned with the known bevacizumab safety profile. These results support first-line bevacizumab + CP therapy in Chinese patients with ovarian cancer. 
		                        		
		                        		
		                        		
		                        	
6.The efficiency and safety of percutaneous nephrolithotomy with negative pressure suction sheath in the treatment of infectious renal calculus
Yi YANG ; Jieqing CHEN ; Jianli CHENG ; Xiaohong HAN ; Jiou LI ; Xuezi ZHENG ; Hongbing MEI
Chinese Journal of Urology 2023;44(12):922-926
		                        		
		                        			
		                        			Objective:To compare the efficacy and safety of percutaneous nephrolithotomy (PCNL) with negative pressure suction sheath and PCNL with traditional expanded sheath in the treatment of infectious renal calculus.Methods:From May 2019 to June 2022 in our department, 35 patients with infectious renal calculus who received PCNL with negative pressure suction sheath (negative pressure sheath group, NPS group) and another 35 patients with infectious renal calculus who received PCNL with traditional expanded sheath (control group) were determined in our research. Propensity score matching (PSM) was conducted. Preoperative clinical data of the 2 groups was similar and there were no statistical differences between the 2 groups in the age [(45.5±6.8)vs. (44.9±7.3) years old, P=0.723], gender (man/woman 19/16 cases vs. 21/14 cases, P=0.629), body mass index(BMI) [(24.2±4.2)kg/m 2vs. (24.5±3.9)kg/m 2, P=0.758], American Society of Anesthesiologists risk score(ASA) (grade 1/grade 2: 30/5 cases vs. 29/6 cases, P=0.743), sides of calculus (left/right: 18/17 cases vs. left 17/18 cases, P=0.811), Guy’s stone score (grade Ⅰ/Ⅱ/Ⅲ: 3/25/7 cases vs. 1/29/5 cases, P=0.443), CT value of calculus [(629.2±98.8)HU vs. (608.5±105.1)HU, P=0.399], urinary leucocyte (-/+ /+ + : 29/5/1 cases vs. 28/5/2 cases, P=0.839), hypertension(3 cases vs. 5 cases, P=0.707), diabetes(2 cases vs. 2 cases, P=1.000). The ureteral catheter on the affected side was indwelled in the lithotomy position, and ultrasound guided positioning puncture was performed on the affected renal side of the posterior axillary line in the prone position. The puncture channel was established and then expanded to F20 successively, and the lithotriptic sheath was placed to establish the lithotriptic channel. Compared with the traditional expanded sheath, the negative pressure suction sheath was different in that the collateral suction channel was added on the main gravel channel and connected with continuous negative pressure suction. The negative pressure was 40 kPa. All patients were treated with pneumatic ballistic lithotripsy combined with holmium laser. KUB was performed within 1 week after surgery. We defined stone removal as either no residual stones or clinically insignificant residual stones (≤4 mm) which did not cause urinary obstruction. The intraoperative duration of operation and postoperative clinical parameters [white blood cell(WBC), procalcitonin(PCT), C-reactive protein(CRP), hemoglobin(Hb), stone clearance rate] and incidence of perioperative complications were compared between the 2 groups. Results:The operation time of NPS group was lower than that in control group [(35.6±19.5)min vs. (45.4±20.2)min, P<0.05]. The proportion of patients with increased WBC, PCT and CRP in blood after operation in NPS group was lower than that in control group, and there were (WBC: 25.7% vs. 54.3%, P<0.05), (PCT: 42.9% vs.68.6%, P<0.05) and (CRP: 62.9% vs.85.7%, P<0.05) respectively. There was no significant difference in the proportion of patients with decreased Hb postoperatively between the 2 groups (2.9% vs. 8.6%, P=0.607). There was no significant difference in calculus clearance rate postoperatively between the 2 groups (97.1% vs. 94.3%, P=1.000). Postoperative calculus component analysis of the 2 groups suggested that all patients had infected calculus dominated by ammonium magnesium phosphate and phosphate apatite. The incidence of perioperative complications in NPS group was lower than that in control group (22.9% vs. 51.4%, P<0.05). The proportion of patients with fever (body temperature>37.5℃) postoperatively in NPS group was lower than that in control group (14.3% vs. 37.1%, P<0.05). There were 2 and 3 patients respectively required upgraded antibiotic therapy after operation in the 2 groups ( P=1.000). There was one patient respectively with urinary tract obstruction and renal colic due to blood clots postoperatively in each group ( P=1.000). There was one patient with urinary sepsis in control group after operation ( P=0.476). Conclusions:Compared with PCNL with traditional expanded sheath, PCNL with negative pressure suction sheath can save operation time for infectious renal calculus, and reduce the incidence of postoperative infection and perioperative complications. Therefore, the safety of negative pressure suction sheath is higher. However, there is no difference in stone clearance rate between them.
		                        		
		                        		
		                        		
		                        	
7.Molecular basis and homeostatic regulation of Zinc taste.
Rui LUO ; Yuxiang ZHANG ; Yinjun JIA ; Yan ZHANG ; Zongyang LI ; Jieqing ZHAO ; Ting LIU ; Wei ZHANG
Protein & Cell 2022;13(6):462-469
8.Histopathological and MRI characteristics of facial unidentified filling objects
Lei DONG ; Li QUAN ; Wencai WENG ; Weiyan CHEN ; Jieqing WANG ; Wei YANG ; Xu HAN ; Bo HU ; Xiuxiu ZHANG ; Chen ZHANG
Chinese Journal of Medical Aesthetics and Cosmetology 2021;27(2):124-128
		                        		
		                        			
		                        			Objective:Unidentified filling objects (UFO) can cause adverse results including infections, overfilling, asymmetry, foreign body granulomas, dislocation or psychological panic. To remove UFO accurately, it is important to locate and identify the injected substances preoperatively. This study investigated the viability of using MRI to correctly locate and identify injected substances by relating MRI to gross and pathological microscopic examination.Methods:Eighty-two facial UFO patients from 2013 to 2017 were studied by the experts of the Department of Image, Xinhua Hospital of Dalian University. Five of the patients were male and seventy-seven were female. The age ranged from 17 to 58 years with average 29.4 years. They came to our hospital for removal of UFO after they had facial injective fillers in the illegal medial offices. The injected sites involved in the forehead, temple, malar, cheek, nose, nasolabial folds, and chin. All the patients' faces were examined with MRI preoperatively, using T1W, T2W and fat-suppressed sequences. Based on the guides of MRI, UFOs were removed with their capsules by open approach. Samples were recorded with digital pictures and then were fixed in 10% formalin solution for microscopic examination of HE stained slices.Results:Based on MRI, gross and microscopic examination, UFO were classified into 3 types, gel-like fillers, solid particles, and growth factors. Gel-like fillers appeared strongly hyperintense on T2 W and STIR sequences and hypointense on T1 W sequences. Grossly, they looked like gruel covered by altered soft tissue. Under the light microscope, many pieces of blue-stained material were dispersed in subcutaneous tissue infiltrated with a large number of mononuclear cells and foreign-body giant cells. Solid particles had low to intermediate signal intensity on T1 W and T2 W images. Grossly, they were like sand merging in soft tissue. The biopsy showed crowed bubbles surrounded by tissue filled with a large number of mononuclear cells and foreign-body giant cells. For growth factors affected tissue, it was hard to differentiate between normal and abnormal on MRI. The affected tissue appeared as somehow hypointense on T1 W sequences and hyperintense on T2 W fat suppressed sequences. During the operation, the affected region was easy bleeding and full of fibrofatty tissue. Under the microscope, there were increased small blood vessels and collagens.Conclusions:Based on MRI, gross and microscopic examination, UFO can be classified into 3 types, gel-like fillers, solid particles, and growth factors. MRI is very important for doctors to assess the patient's conditions and make the plan of operation. MRI is also useful for doctors to locate UFO and understand the relationship between UFO and their nearby organs.
		                        		
		                        		
		                        		
		                        	
9.The expression of clock gene CLOCK and its clinical significance in nasopharyngeal carcinoma
Xiaomei LI ; Yuanyuan LI ; Chaofen ZHAO ; Lina LIU ; Qianyong HE ; Jieqing JIANG ; Yue CHEN ; Minghui YANG ; Yaxue TANG ; Yuxin LI ; Feng JIN
Chinese Journal of Oncology 2021;43(12):1255-1263
		                        		
		                        			
		                        			Objective:To explore the relationship between expression levels of CLOCK mRNA and protein and the clinical characteristics of patients with nasopharyngeal carcinoma.Methods:The frozen tissue specimens from 33 patients with nasopharyngeal carcinoma in the Affiliated Tumor Hospital of Guizhou Medical University from 2018 to 2019 were collected. Seventeen cases of tissue specimens from patients with nasopharyngeal chronic inflammation in the Affiliated Hospital of Guizhou Medical University in 2019 were collected. From 2008 to 2014, 68 cases of formalin-fixed paraffin-embedding (FFPE) nasopharyngeal carcinoma tissue and 37 cases of FFPE nasopharyngeal chronic inflammation tissue were collected from the Affiliated Tumor Hospital of Guizhou Medical University. Real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blot (WB) were used to detect the mRNA and protein expression levels of CLOCK. The nasopharyngeal carcinoma cells including CNE1, CNE2, 5-8F and the normal nasopharyngeal epithelial cell NP69 were cultured. qRT-PCR was used to detect the expression level of CLOCK mRNA in each cell line at the time points of ZT2, ZT6, ZT10, ZT14, ZT18 and ZT22. The cosine method was used to fit the rhythm of CLOCK gene in nasopharyngeal carcinoma. The protein expression of CLOCK protein was detected by using immunohistochemical method in 68 cases of nasopharyngeal carcinoma and 37 cases of nasopharyngeal chronic inflammation tissue. Survival was analyzed by Kaplan-Meier method and Log rank test, and the influencing factors was analyzed by Cox regression model.Results:The expression levels of CLOCK mRNA in CNE1, CNE2 and 5-8F cells (0.63±0.07, 0.91±0.02 and 0.33±0.04, respectively) were lower than that in NP69 cell (1.00±0.00, P<0.05). The expression levels of CLOCK protein in CNE1, CNE2 and 5-8F cells (0.79±0.06, 0.57±0.05 and 0.74±0.10, respectively) were lower than that of NP69 cells (1.00±0.00, P<0.05). The expressions of CLOCK mRNA in nasopharyngeal carcinoma cells including CEN1, CNE2, 5-8F and normal nasopharyngeal epithelial cell NP69 were different at different time points, with temporal fluctuations. The fluctuation periods of CLOCK mRNA in CNE1, CNE2, 5-8F, and NP69 cells were 16, 14, 22 and 24 hours, respectively. The peak and trough times were ZT10: 40 and ZT18: 40, ZT10 and ZT3, ZT14: 30 and ZT3: 30, ZT12: 39 and ZT0: 39, respectively. CLOCK mRNA and protein expression levels in nasopharyngeal carcinoma tissues (0.37±0.20 and 0.20±0.26, respectively) were lower than those in nasopharyngeal chronic inflammation tissues (1.00±0.00 and 0.51±0.41, respectively, P<0.05). The 1, 3, and 5-year survival rates of patients in the CLOCK protein high expression group (CLOCK protein expression level ≥ 0.178) were 96.2%, 92.1%, and 80.1%, respectively, which were higher than those in the low expression group (CLOCK protein expression level <0.178, 92.9% , 78.6% and 57.1%, respectively, P=0.009). The 1, 3, and 5-year progression-free survival (PFS) rates of patients in the CLOCK protein high expression group were 96.2%, 87.8%, and 87.7%, respectively, which were higher than those in the low expression group (92.7%, 82.2%, and 70.8%, respectively, P=0.105). Compared with the low-expression group (100.0%, 96.9%, and 90.0%, respectively), the 1, 3, and 5-year recurrence-free survival rates of patients in the CLOCK protein high expression group (100.0%, 95.7%, and 95.7%, respectively) were not statistically significant ( P=0.514). Compared with the low-expression group (92.7%, 82.2%, and 79.3%), the 1, 3, and 5-year survival rates without metastasis in the CLOCK protein high expression group (96.2%, 92.0%, and 92.0%, respectively) were not statistically significant ( P=0.136). CLOCK protein expression and T stage were independent prognostic factors of overall survival ( P<0.05). Conclusions:The expression of CLCOK is downregulated in the nasopharyngeal carcinoma cell and nasopharyngeal carcinoma tissues. Clock gene CLOCK is rhythmically expressed in the nasopharyngeal carcinoma cells and normal nasopharyngeal epithelial cells. Compared with normal nasopharyngeal epithelial cells, the fluctuation period of CLOCK in nasopharyngeal carcinoma cells is shortened. The overall survival of patients in the CLOCK protein high expression group is better than that of low expression group. The expression of CLOCK protein is an independent influencing factor for overall survival. CLOCK gene may be a potential tumor suppressor gene in the nasopharyngeal carcinoma.
		                        		
		                        		
		                        		
		                        	
10.The expression of clock gene CLOCK and its clinical significance in nasopharyngeal carcinoma
Xiaomei LI ; Yuanyuan LI ; Chaofen ZHAO ; Lina LIU ; Qianyong HE ; Jieqing JIANG ; Yue CHEN ; Minghui YANG ; Yaxue TANG ; Yuxin LI ; Feng JIN
Chinese Journal of Oncology 2021;43(12):1255-1263
		                        		
		                        			
		                        			Objective:To explore the relationship between expression levels of CLOCK mRNA and protein and the clinical characteristics of patients with nasopharyngeal carcinoma.Methods:The frozen tissue specimens from 33 patients with nasopharyngeal carcinoma in the Affiliated Tumor Hospital of Guizhou Medical University from 2018 to 2019 were collected. Seventeen cases of tissue specimens from patients with nasopharyngeal chronic inflammation in the Affiliated Hospital of Guizhou Medical University in 2019 were collected. From 2008 to 2014, 68 cases of formalin-fixed paraffin-embedding (FFPE) nasopharyngeal carcinoma tissue and 37 cases of FFPE nasopharyngeal chronic inflammation tissue were collected from the Affiliated Tumor Hospital of Guizhou Medical University. Real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blot (WB) were used to detect the mRNA and protein expression levels of CLOCK. The nasopharyngeal carcinoma cells including CNE1, CNE2, 5-8F and the normal nasopharyngeal epithelial cell NP69 were cultured. qRT-PCR was used to detect the expression level of CLOCK mRNA in each cell line at the time points of ZT2, ZT6, ZT10, ZT14, ZT18 and ZT22. The cosine method was used to fit the rhythm of CLOCK gene in nasopharyngeal carcinoma. The protein expression of CLOCK protein was detected by using immunohistochemical method in 68 cases of nasopharyngeal carcinoma and 37 cases of nasopharyngeal chronic inflammation tissue. Survival was analyzed by Kaplan-Meier method and Log rank test, and the influencing factors was analyzed by Cox regression model.Results:The expression levels of CLOCK mRNA in CNE1, CNE2 and 5-8F cells (0.63±0.07, 0.91±0.02 and 0.33±0.04, respectively) were lower than that in NP69 cell (1.00±0.00, P<0.05). The expression levels of CLOCK protein in CNE1, CNE2 and 5-8F cells (0.79±0.06, 0.57±0.05 and 0.74±0.10, respectively) were lower than that of NP69 cells (1.00±0.00, P<0.05). The expressions of CLOCK mRNA in nasopharyngeal carcinoma cells including CEN1, CNE2, 5-8F and normal nasopharyngeal epithelial cell NP69 were different at different time points, with temporal fluctuations. The fluctuation periods of CLOCK mRNA in CNE1, CNE2, 5-8F, and NP69 cells were 16, 14, 22 and 24 hours, respectively. The peak and trough times were ZT10: 40 and ZT18: 40, ZT10 and ZT3, ZT14: 30 and ZT3: 30, ZT12: 39 and ZT0: 39, respectively. CLOCK mRNA and protein expression levels in nasopharyngeal carcinoma tissues (0.37±0.20 and 0.20±0.26, respectively) were lower than those in nasopharyngeal chronic inflammation tissues (1.00±0.00 and 0.51±0.41, respectively, P<0.05). The 1, 3, and 5-year survival rates of patients in the CLOCK protein high expression group (CLOCK protein expression level ≥ 0.178) were 96.2%, 92.1%, and 80.1%, respectively, which were higher than those in the low expression group (CLOCK protein expression level <0.178, 92.9% , 78.6% and 57.1%, respectively, P=0.009). The 1, 3, and 5-year progression-free survival (PFS) rates of patients in the CLOCK protein high expression group were 96.2%, 87.8%, and 87.7%, respectively, which were higher than those in the low expression group (92.7%, 82.2%, and 70.8%, respectively, P=0.105). Compared with the low-expression group (100.0%, 96.9%, and 90.0%, respectively), the 1, 3, and 5-year recurrence-free survival rates of patients in the CLOCK protein high expression group (100.0%, 95.7%, and 95.7%, respectively) were not statistically significant ( P=0.514). Compared with the low-expression group (92.7%, 82.2%, and 79.3%), the 1, 3, and 5-year survival rates without metastasis in the CLOCK protein high expression group (96.2%, 92.0%, and 92.0%, respectively) were not statistically significant ( P=0.136). CLOCK protein expression and T stage were independent prognostic factors of overall survival ( P<0.05). Conclusions:The expression of CLCOK is downregulated in the nasopharyngeal carcinoma cell and nasopharyngeal carcinoma tissues. Clock gene CLOCK is rhythmically expressed in the nasopharyngeal carcinoma cells and normal nasopharyngeal epithelial cells. Compared with normal nasopharyngeal epithelial cells, the fluctuation period of CLOCK in nasopharyngeal carcinoma cells is shortened. The overall survival of patients in the CLOCK protein high expression group is better than that of low expression group. The expression of CLOCK protein is an independent influencing factor for overall survival. CLOCK gene may be a potential tumor suppressor gene in the nasopharyngeal carcinoma.
		                        		
		                        		
		                        		
		                        	
            
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