1.Mechanism by which KRT18 interacts with mRNA and long non-coding RNA to regulate intervertebral disc nucleus pulposus cell injury
Zhongyuan LIU ; Yang LI ; Zhiwen ZHANG
Chinese Journal of Tissue Engineering Research 2025;29(2):312-321
		                        		
		                        			
		                        			BACKGROUND:Differentially expressed RNA-binding proteins in the intervertebral disc plays a key role in intervertebral disc degeneration,and decreased levels of the RNA-binding protein KRT18 are associated with degenerative disc disease,but its specific role in the nucleus pulposus cells has not yet been fully determined. OBJECTIVE:To investigate the interaction of KRT18 with mRNA and long non-coding RNA on nucleus pulposus cells of the intervertebral disc and its mechanism. METHODS:Normal and degenerated nucleus pulposus cells were obtained from nucleus pulposus samples of patients undergoing interbody fusion for lumbar fracture or intervertebral disc degeneration.iRIP-seq,functional enrichment analysis,and DNA microarray analysis were performed to identify the mRNA and long non-coding RNA binding with KRT18.Subsequently,KRT18 was knocked down in nucleus pulposus cells based on the analysis results,and the expression levels of related genes were detected at the protein and RNA levels through protein immunoblotting and qRT-PCR,respectively. RESULTS AND CONCLUSION:Through iRIP-seq analysis,we identified abundant KRT18 binding sites within the GUAAUC and AGCCUC sequences,indicating that KRT18 may be involved in regulating RNA transcription,translation,stability or play a role in cell signaling pathways.It can stably bind to mature mRNA,among which highly expressed genes include CRLF1,IGFBP4,etc.At the same time,the peak genes of long non-coding RNA binding with it include SNHG25,SNHG12,NEAT1,USP32,EIF4A2 and CDH4.Most of these genes are involved in various biological processes such as apoptosis and inflammation,and can mediate related pathways of extracellular matrix metabolism.KRT18 can regulate their stability,transport,translation,splicing and other functions,thus affecting gene expression and cell function.We further verified through experiments the knockdown of KRT18 in nucleus pulposus cells,and found that the level of extracellular matrix metabolism was inhibited and unbalanced,resulting in intervertebral disc degeneration in vitro.This study investigated the regulatory mechanism of KRT18 from the perspective of its binding with mRNA and long non-coding RNA for the first time,and speculated the potential function of KRT18 in the pathogenesis of intervertebral disc degeneration,laying a foundation for future research on the key functions of KRT18.
		                        		
		                        		
		                        		
		                        	
2.Clinical significance of detection of EGFR mutation in alveolar lavage fluid exosome of patients with advanced non-small cell lung cancer
Changheng YANG ; Ying CHEN ; Zhongyuan ZHANG ; Qingqing MA
The Journal of Practical Medicine 2024;40(1):48-52
		                        		
		                        			
		                        			Objective To compare and analyze the gene mutation of EGFR of bronchoalveolar lavage fluid(BALF)exosome,serum and lung cancer tissue specimens of patients with advanced non-small cell lung cancer(NSCLC)and assess whether the BALF exosome specimens are suitable for screening before clinical targeted therapy,to provide new ideas and screening methods for early individualized treatment of advanced NSCLC patients.Methods BALF exosomes,serum and lung cancer tissue specimens EGFR gene mutations of 78 cases with advanced NSCLC were detected by using amplification refractory mutation system(ARMS)method in Department of Respiratory and Critical Care Medicine in our hospital from May 2021 to May 2023,and the results were retrospec-tively analyzed.A comparative analysis of the specimens was conducted using lung cancer tissue specimens as bench-marks.Results A total of 33,25 and 38 cases of EGFR gene mutation and 42,53 and 40 cases of EGFR wild type were detected in BALF exosomes,serum and lung cancer tissues specimens respectively.The mutation rate of EGFR gene was 42.3%(33/78,32.1%(25/78)and 48.7%(38/78)in BALF exosomes,serum and lung cancer tissues specimens respectively.EGFR detection showed no results in 3 cases and the false-negative rate was 6.4%(5/78)in BALF specimen,and false-negative rate was 16.7%(13/78)in serum.The detection coincidence rate of EGFR mutation was 86.8%(33/38)in BALF exosomes specimen,and 65.8%(25/38)in serum.Conclusions EGFR gene mutation rate in BALF exosome specimen is consistent with that in serum and lung cancer tissue samples,showing no statistical significance(P>0.05).It is superior to serum specimen and suitable for patient screening before targeted therapy and provides new ideas and screening methods for early individualized treatment decisions of advanced NSCLC patients.
		                        		
		                        		
		                        		
		                        	
3.Analysis of the efficacy of arthroscopic transverse release of iliotibial band through peritrochanteric space for the treatment of external snapping hip
Yidong WU ; Kangkang YU ; Zhongyao LI ; Lu GAN ; Qi JIA ; Zhongyuan ZHAO ; Yang HE ; Zhikai GUO ; Chunbao LI
Chinese Journal of Orthopaedics 2024;44(1):18-24
		                        		
		                        			
		                        			Objective:To analyze the clinical efficacy of arthroscopic transverse release of the iliotibial band through peritrochanteric space for the treatment of external snapping hip.Methods:A total of 30 patients (12 males and 18 females) with bilateral external snapping hip underwent arthroscopic transverse release of the iliotibial band through peritrochanteric space in Department of Sports Medicine, Senior Department of Orthopaedics, the Fourth Medical Center, Chinese PLA General Hospital were retrospectively analyzed from May 2021 and June 2022. The average age was 32.5±8.2 years (range, 17-51 years). At the same time, 30 patients who underwent arthroscopic external release of the iliotibial band through the external surface of the iliotibial band (external iliotibial band group) were selected as control group, including 13 males and 17 females, aged 29.5±6.8 years (range, 11-45 years). The visual analogue scale (VAS), modified Harris hip score (mHHS), and gluteal muscle contracture disability scale (GDS) were compared between the two groups at preoperative, 6 months postoperative, and final follow-up.Results:All patients successfully completed the operation and were followed up for 17.5±3.3 months (range, 12-25 months). The VAS scores of the two groups at the last follow-up were lower than those before operation ( P<0.05). The mHHS scores before operation, 6 months after operation and at the last follow-up in the peritrochanteric space group were 76.5 (67.0, 85.5), 98.5 (94.8, 100.0) and 100.0 (97.0, 100.0), respectively, and those in the external iliotibial band group were 80.5 (70.0, 86.0), 100.0 (96.0, 100.0) and 100.0 (99.5, 100.0). The differences in mHHS scores between the two groups were statistically significant for intragroup comparisons ( P<0.05); of these, 6 months postoperatively and at the last follow-up were greater than preoperatively, with statistically significant differences ( P<0.05); the differences at 6 months postoperatively and at the last follow-up were not statistically significant ( P>0.05). There was no significant difference in mHHS scores between groups at different time points ( P>0.05). The GDS before operation, at 6 months after operation and at the last follow-up were 47.0 (35.8, 64.5), 90.0 (81.0, 94.0) and 93.5 (89.8, 98.0) in the peritrochanteric space group, and 51.0 (38.0, 64.5), 50.0 (81.0, 94.0) and 93.5 (89.8, 98.0) in the external iliotibial band group, respectively. The differences in GDS between the two groups were statistically significant for intragroup comparisons ( P< 0.05); of these, 6 months postoperatively and at the last follow-up were greater than preoperatively, with statistically significant differences ( P<0.05); the differences at 6 months postoperatively and at the last follow-up were not statistically significant ( P>0.05). There was no significant difference in GDS between groups at different time points ( P>0.05). Conclusion:Arthroscopic transverse release of the iliotibial band through peritrochanteric space for the treatment of external snapping hip can effectively reduce hip pain and improve hip function, with satisfactory clinical results, and can be used as an alternative treatment to transverse release through the external surface of the iliotibial band.
		                        		
		                        		
		                        		
		                        	
4.Awareness Investigation and Strategy Analysis on Pharmaceutical Services and Fees in Hospitals in Guizhou Province
Dongmei LI ; Qian YANG ; Shuimei SUN ; Ling HE ; Dirong WU ; Mingji LIU ; Pingping CHEN ; Libin WANG ; Zhongyuan WANG ; Hong ZHANG ; Zhu ZHU ; Xue BAI ; Changcheng SHENG ; Jiaxing ZHANG ; Lei LU ; Xue WANG ; Qi CHEN
Herald of Medicine 2024;43(9):1410-1415
		                        		
		                        			
		                        			Objective To investigate the current status and awareness of pharmaceutical services in hospitals in Guizhou province and to provide a reference for exploring and carrying out pharmaceutical service fees.Methods The questionnaire was designed by the"wjx.cn"website.Three kinds of questionnaires were designed for pharmacists,doctors,nurses,and patients as the research objects,with corresponding differences in some questions,and promoted on WeChat,Dingxiangyuan,and other network platforms.Results A total of 655 questionnaires were collected,and 639 valid questionnaires were recovered,with an effective recovery rate of 97.56%.324 pharmacists(50.70%),82 doctors and nurses(12.83%),233 patients(36.46%)were surveyed.The average approval score of these three groups of respondents on pharmaceutical service fees was 4.67,4.23,and 4.22,respectively(full score:5).Conclusions Overall,pharmacists'professional services have received support from medical staff and patients.However,patients'pharmaceutical service projects currently focus on dispensing services.The recognition of pharmacists'work and the public's awareness of pharmaceutical services can be improved by enhancing the professional ability of pharmacists,strengthening publicity and guidance,and exploring"Internet+pharmaceutical services",etc.,to promote the sustainable development of pharmaceutical services.
		                        		
		                        		
		                        		
		                        	
5.Investigation of pharmacodynamic material basis of Schisandra chinensis in the treatment of allergic asthma
Yifan BING ; Tianlei ZHANG ; Zhiwei SUN ; Xiaolong YANG ; Sunan LI ; Xue JIANG ; Zhongyuan QU
China Pharmacy 2023;34(3):315-320
		                        		
		                        			
		                        			OBJECTIVE To study the pharmacological basis of Schisandra chinensis in the treatment of allergic asthma. METHODS The common components of 10 batches of S. chinensis from different habitats were analyzed by UPLC-Q-TOF-MS/MS. Furthermore, the allergic asthma model was established by intraperitoneal injection of ovalbumin (OVA) and aluminum hydroxide for stimulation combined with atomization exitation; general behavioral observation and the contents of interferon γ (IFN-γ), interleukin-4 (IL-4) and immunoglobulin E (IgE) in serum were taken as criteria for evaluating the therapeutic effect of S. chinensis from different habitats in the treatment of allergic asthma. Correlation coefficients between common peak area and efficacy evaluation index of each batch of medicinal material were analyzed through grey correlation degree and Pearson correlation analysis. RESULTS A total of 21 common components were identified in 10 batches of S. chinensis from different habitats. After administration of S. chinensis, symptoms such as shortness of breath, sneezing and curling of rats were alleviated. In addition, the content of IFN-γ was significantly increased while the contents of IL-4 and IgE in serum were distinctly decreased (P<0.01). Grey correlation analysis showed that 11 common components had high correlation coefficients with IFN-γ, IL-4 and IgE (rˉ>0.8). Pearson correlation analysis showed that 8 components were significantly positively correlated with the content of IFN-γ (P< 0.05), and 9, 8 components were significantly negatively correlated with the content of IL-4 and IgE (P<0.05). Based on the results of grey correlation degree and Pearson correlation analysis, 7 components such as peak 3, 4, 6, 7, 9, 19 and 20, were highly related to S. chinensis in the treatment of allergic asthma. CONCLUSIONS Schisandrol A, schisandrin B, schisandrin C, gomisin M2, gomisin J, pregomisin and angeloylgomisin H are the potential pharmacodynamic substance basis of S. chinensis in the treatment of allergic asthma.
		                        		
		                        		
		                        		
		                        	
6.Development of a flexible embedded neurostimulator for animal robots.
Zhenling SU ; Dongyun WANG ; Xiaomin QI ; Chenguang YANG ; Yexin ZHANG ; Kaige LIU ; Yue QIN ; Xinyu LIU
Journal of Biomedical Engineering 2023;40(2):327-334
		                        		
		                        			
		                        			The neural stimulator is a core component of animal robots. While the control effect of animal robots is influenced by various factors, the performance of the neural stimulator plays a decisive role in regulating animal robots. In order to optimize animal robots, embedded neural stimulators had been developed using flexible printed circuit board technology. This innovation not only enabled the stimulator to generate parameter-adjustable biphasic current pulses through control signals, but also optimized its carrying mode, material, and size, overcoming the disadvantages of traditional backpack or head-inserted stimulators, which have poor concealment and are prone to infection. Static, in vitro, and in vivo performance tests of the stimulator demonstrated that it not only had precise pulse waveform output capability, but also was lightweight and small in size. It had excellent in vivo performance in both laboratory and outdoor environments. Our study has high practical significance for the application of animal robots.
		                        		
		                        		
		                        		
		                        			Animals
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		                        			Robotics
		                        			
		                        		
		                        	
7.Optimized CRISPR/Cas9 System in Brain Science and Application Prospects in Field of Traditional Chinese Medicine
Shuoqiu DENG ; Shuiqing QU ; Yu ZHANG ; Yuanmin YANG ; Zhongyuan ZHENG ; Tuo LIU ; Lina CHEN ; Yujie LI
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(1):169-180
		                        		
		                        			
		                        			Clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR associated nuclease 9 (CRISPR/Cas9) is a self-defense system found in bacteria and archaea that enables targeted gene editing based on the principle. Due to its universality, efficiency, and simplicity, CRISPR/Cas9 has been applied in the pathological mechanism and prevention and treatment of diseases in many fields. Cerebrovascular diseases and central nervous system diseases seriously endanger human health. Stroke is related to genetics, unhealthy living habits, chronic diseases, and other factors. The brain tissue structure is complex and the cell types are diverse. It is difficult for a universal gene editing platform to study target genes safely, specifically, and efficiently. Scholars have continuously improved and optimized gene editing technology, explored the potential and research methods of gene editing technology, and promoted the research process of brain science. After a brief introduction to the mechanism of CRISPR/Cas9, this paper mainly summarized the optimization of the system in the fields of cerebral science including delivery methods, adeno-associated virus assembly, and new nanomaterials. Its application in cerebrovascular research including vascular homeostasis, microglial homeostasis, angiogenesis, blood-brain barrier, and drug screening was also summarized. Finally, this paper prospected the development of CRISPR/Cas9 in traditional Chinese medicine, hoping to provide references for related research design. 
		                        		
		                        		
		                        		
		                        	
8.Clinicopathological comparison of intrahepatic cholangiocarcinoma with and without viral hepatitis
Sen YANG ; Yanbing LIU ; Pan WU ; Shuai YAN ; Feilong TAN ; Zhongyuan ZHAO ; Yiyang YAN ; Xueliang YUE ; Hongshan LIU
Chinese Journal of General Surgery 2023;38(4):292-296
		                        		
		                        			
		                        			Objective:To analyze the different clinicopathological features of intrahepatic cholangiocarcinoma with and without viral hepatitis.Methods:The clinicopathological data of 79 intrahepatic cholangiocarcinoma cases from Mar 2012 to Sep 2018 at Henan Provincial People's Hospital were retrospectively analyzed.Results:Twenty-five of the 79 patients with intrahepatic cholangiocarcinoma were accompanied by viral hepatitis. Those with viral hepatitis had a lower mean age at onset than those without [(53±11) years vs. (60±11) years, P=0.011], higher proportion of male patients (80% vs. 52%, P=0.017), higher AFP positive rate (40% vs. 19%, P=0.041), lower CA19-9 positive rate (48% vs. 72%, P=0.036), tend to occur in the right liver lobe (76% vs. 44%, P=0.009), a lower rate of bile duct invasion (16% vs. 41%, P=0.03), and were more likely to be mass type (mass type proportion 96% vs. 72%, P=0.032). Conclusions:Viral hepatitis is common in intrahepatic cholangiocarcinoma. Intrahepatic cholangiocarcinoma with and without viral hepatitis differ in clinicopathology. Intrahepatic cholangiocarcinoma with viral hepatitis is more likely to have the characteristics of hepatocellular carcinoma, while intrahepatic cholangiocarcinoma without viral hepatitis is more likely to have the characteristics of cholangiocarcinoma.
		                        		
		                        		
		                        		
		                        	
9.Robotic urologic surgery using the KangDuo-Surgical Robot-01 system: A single-center prospective analysis.
Shengwei XIONG ; Shubo FAN ; Silu CHEN ; Xiang WANG ; Guanpeng HAN ; Zhihua LI ; Wei ZUO ; Zhenyu LI ; Kunlin YANG ; Zhongyuan ZHANG ; Cheng SHEN ; Liqun ZHOU ; Xuesong LI
Chinese Medical Journal 2023;136(24):2960-2966
		                        		
		                        			BACKGROUND:
		                        			The KangDuo-Surgical Robot-01 (KD-SR-01) system is a new surgical robot recently developed in China. The aim of this study was to present our single-center experience and mid-term outcomes of urological procedures using the KD-SR-01 system.
		                        		
		                        			METHODS:
		                        			From August 2020 to April 2023, consecutive urologic procedures were performed at Peking University First Hospital using the KD-SR-01 system. The clinical features, perioperative data, and follow-up outcomes were prospectively collected and analyzed.
		                        		
		                        			RESULTS:
		                        			A total of 110 consecutive patients were recruited. Among these patients, 28 underwent partial nephrectomy (PN), 41 underwent urinary tract reconstruction (26 underwent pyeloplasty, 3 underwent ureteral reconstruction and 12 underwent ureterovesical reimplantation [UR]), and 41 underwent radical prostatectomy (RP). The median operative time for PN was 112.5 min, 157.0 min for pyeloplasty, 151.0 min for ureteral reconstruction, 142.5 min for UR, and 138.0 min for RP. The median intraoperative blood loss was 10 mL for PN, 10 mL for pyeloplasty, 30 mL for ureteral reconstruction, 20 mL for UR, and 50 mL for RP. All procedures were successfully completed without conversion, and there were no major complications in any patient. The median warm ischemia time of PN was 17.3 min, and positive surgical margin was not noted in any patient. The overall positive surgical margin rate of RP was 39% (16/41), and no biochemical recurrence was observed in any RP patient during the median follow-up of 11.0 months. The surgical success rates of pyeloplasty and UR were 96% (25/26) and 92% (11/12) during the median follow-up of 29.5 months and 11.5 months, respectively.
		                        		
		                        			CONCLUSION
		                        			The KD-SR-01 system appears feasible, safe, and effective for most urological procedures, based on our single-center experience.
		                        		
		                        		
		                        		
		                        			Male
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		                        			Humans
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		                        			Robotic Surgical Procedures/methods*
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		                        			Robotics
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		                        			Treatment Outcome
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		                        			Retrospective Studies
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		                        			Ureter/surgery*
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		                        			Urologic Surgical Procedures/methods*
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		                        			Laparoscopy/methods*
		                        			
		                        		
		                        	
10.Clinical value of lymphocyte count in autoimmune diabetic nephropathy
Zhongyuan XIANG ; Wen'en LIU ; Sangya FU ; Jingyi HU ; Yanyi YANG
Journal of Central South University(Medical Sciences) 2023;48(11):1639-1649
		                        		
		                        			
		                        			Objective:In recent years,the prevalence of diabetic nephropathy(DN)has increased significantly.An increasing number of studies have shown that lymphocyte-associated inflammatory responses play a role in DN.This study aims to investigate the relationship between lymphocytes and DN in patients with autoimmune diabetes. Methods:The clinical data of 226 patients with Type 1 diabetes(T1D)and 79 patients with latent autoimmune diabetes in adults(LADA)were retrospectively studied and stratified according to the urinary albumin to creatinine ratio(ACR).Risk factors associated with DN were analyzed using correlation analysis and logistic regression. Results:In T1D and LADA patients,systolic blood pressure(SBP),uric acid duration,and diabetes duration in patients with normoalbuminuria were lower or shorter than those in patients with macroalbuminuria(P<0.05).The lymphocyte count of T1D patients was significantly higher than that in LADA patients(P<0.05),while the neutrophil to lymphocyte ratio(NLR)of T1D patients was significantly lower than that in LADA patients(P<0.05).The lymphocyte count in the T1D patients with normoalbuminuria was lower than that those with macroalbuminuria(P<0.05).The NLR was lower in the T1D patients with macroalbuminuria than those with microalbuminuria and normoproteinuria(all P<0.01).Based on logistic regression analysis,lymphocytes were independently associated with DN in T1D after adjusting for various known risk factors such as course of disease,age,gender,dyslipidemia,hypertension,and smoking status.Analysis of the receiver operating characteristic curve of subjects predicting lymphocytes in normoalbuminuria showed that the area under the curve was 0.601(95% CI 0.510 to 0.693,P=0.039),and when the cutoff value of lymphocytes was 2.332,the sensitivity was 37.0%,and the specificity was 82.5%. Conclusion:Lymphocyte counts in autoimmune diabetic patients are closely associated with DN,suggesting that lymphocyte-mediated inflammation may be involved in the pathogenesis of DN in autoimmune diabetic patients.This study provides a possible perspective for using lymphocytes as a potential biomarker for the early identification of individuals at risk for DN and potential therapeutic targets for DN.
		                        		
		                        		
		                        		
		                        	
            
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