1.Analysis on Clinical Trial Registration Status of TCM for the Treatment of Cervical Spondylosis
Zibo GAO ; Kai CHEN ; Fangzheng LIN ; Yongjin LI ; Dingkun LIN
Chinese Journal of Information on Traditional Chinese Medicine 2025;32(12):53-59
Objective To analyze the current status and development trends of registered clinical trials of TCM for the treatment of cervical spondylosis;To provide references for relevant trials.Methods Clinical trials of TCM for the treatment of cervical spondylosis registered in Chinese Clinical Trial Registry(ChiCTR),International Traditional Medicine Clinical Trial Registry(ITMCTR),and ClinicalTrials.gov from the establishment of the databases to December 31,2024 were retrieved,and the registration characteristic information of the included trials was analyzed.Results A total of 168 registered trials involving 32 665 subjects were included.The number of registered TCM clinical trials for cervical spondylosis showed an overall upward trend;the average implementation cycle of the studies was(20.1±10.7)months;the regions where the research sponsoring institutions were located were mainly in Shanghai,Beijing,and Guangdong;the funding sources were mostly from government finance and institutional funds;a total of 133 studies underwent ethical review;the main study type was interventional research,and the main study design was randomized parallel-controlled trials;the intervention measures were mainly acupuncture,manual therapy,and Chinese materia medica;the main outcome indicators were scale scores;17 studies met the requirements for data sharing and management plans.Conclusion The number of registered TCM clinical trials for cervical spondylosis is generally increasing,involving multiple subtypes of cervical spondylosis and diversified intervention measures.But there are problems such as significant regional differences,uneven methodological quality,and poor data sharing,which should be improved.
2.Extraction and subculture of neural stem cells from mouse embryonic spinal cord:comparison and analysis on advantages and disadvantages of three commonly used digestive enzymes
Dan LUO ; Zhilin GE ; Yonghui HOU ; Wanshun WANG ; Jiheng ZHAN ; Yu HOU ; Dingkun LIN ; Shudong CHEN
Chinese Journal of Tissue Engineering Research 2025;29(31):6609-6615
BACKGROUND:In the research and application of neural stem cells,cell culture and passage are key links,which directly affect the quality of cells and experimental results.It is of great significance to find the most suitable digestive enzymes that can maintain the biological characteristics of embryonic mouse spinal cord neural stem cells and enhance their passage efficiency.OBJECTIVE:To explore the most suitable digestive enzyme for passage of neural stem cells from the spinal cord of embryonic mice.METHODS:Microscopic dissection was used to isolate and extract spinal cord tissue from E14 d embryonic mice,which was cultured in DMEM/F12 serum-free medium containing epidermal growth factor,basic fibroblast growth factor,and B27.After spherulation,Nestin and Sox2 immunofluorescence identification was performed.During neural stem cell passage and culture,single-cell suspensions were prepared using trypsin,papain,and TrypLETM Express enzyme digestion combined with blow molding.The cell dispersion and spheroidization were observed,and passage 3 cells were stained with propidium iodide to detect cell death.Cell proliferation was detected by counting the total number of cells.Immunofluorescence staining,western blot assay and RT-PCR were used to detect the expression of Olig2,Tuj1,GFAP,and NeuN at the protein and mRNA levels and to identify cell differentiation.RESULTS AND CONCLUSION:After 72 hours of culture,E14 d embryonic mouse spinal cord tissue cells could form suspended neurospheres,which could be passaged after 5-7 days.Compared with trypsin and papain,TrypLETM Express enzyme combined with blow beating method was used for passage.The cell dispersion rate was high,the activity was good,and more NeuN-and Tuj1-positive neurons differentiated.This study optimized the culture and passaging process of neural stem cells,laying a foundation for further research on stem cell transplantation therapy for spinal cord diseases.
3.Extraction and subculture of neural stem cells from mouse embryonic spinal cord:comparison and analysis on advantages and disadvantages of three commonly used digestive enzymes
Dan LUO ; Zhilin GE ; Yonghui HOU ; Wanshun WANG ; Jiheng ZHAN ; Yu HOU ; Dingkun LIN ; Shudong CHEN
Chinese Journal of Tissue Engineering Research 2025;29(31):6609-6615
BACKGROUND:In the research and application of neural stem cells,cell culture and passage are key links,which directly affect the quality of cells and experimental results.It is of great significance to find the most suitable digestive enzymes that can maintain the biological characteristics of embryonic mouse spinal cord neural stem cells and enhance their passage efficiency.OBJECTIVE:To explore the most suitable digestive enzyme for passage of neural stem cells from the spinal cord of embryonic mice.METHODS:Microscopic dissection was used to isolate and extract spinal cord tissue from E14 d embryonic mice,which was cultured in DMEM/F12 serum-free medium containing epidermal growth factor,basic fibroblast growth factor,and B27.After spherulation,Nestin and Sox2 immunofluorescence identification was performed.During neural stem cell passage and culture,single-cell suspensions were prepared using trypsin,papain,and TrypLETM Express enzyme digestion combined with blow molding.The cell dispersion and spheroidization were observed,and passage 3 cells were stained with propidium iodide to detect cell death.Cell proliferation was detected by counting the total number of cells.Immunofluorescence staining,western blot assay and RT-PCR were used to detect the expression of Olig2,Tuj1,GFAP,and NeuN at the protein and mRNA levels and to identify cell differentiation.RESULTS AND CONCLUSION:After 72 hours of culture,E14 d embryonic mouse spinal cord tissue cells could form suspended neurospheres,which could be passaged after 5-7 days.Compared with trypsin and papain,TrypLETM Express enzyme combined with blow beating method was used for passage.The cell dispersion rate was high,the activity was good,and more NeuN-and Tuj1-positive neurons differentiated.This study optimized the culture and passaging process of neural stem cells,laying a foundation for further research on stem cell transplantation therapy for spinal cord diseases.
4.Analysis on Clinical Trial Registration Status of TCM for the Treatment of Cervical Spondylosis
Zibo GAO ; Kai CHEN ; Fangzheng LIN ; Yongjin LI ; Dingkun LIN
Chinese Journal of Information on Traditional Chinese Medicine 2025;32(12):53-59
Objective To analyze the current status and development trends of registered clinical trials of TCM for the treatment of cervical spondylosis;To provide references for relevant trials.Methods Clinical trials of TCM for the treatment of cervical spondylosis registered in Chinese Clinical Trial Registry(ChiCTR),International Traditional Medicine Clinical Trial Registry(ITMCTR),and ClinicalTrials.gov from the establishment of the databases to December 31,2024 were retrieved,and the registration characteristic information of the included trials was analyzed.Results A total of 168 registered trials involving 32 665 subjects were included.The number of registered TCM clinical trials for cervical spondylosis showed an overall upward trend;the average implementation cycle of the studies was(20.1±10.7)months;the regions where the research sponsoring institutions were located were mainly in Shanghai,Beijing,and Guangdong;the funding sources were mostly from government finance and institutional funds;a total of 133 studies underwent ethical review;the main study type was interventional research,and the main study design was randomized parallel-controlled trials;the intervention measures were mainly acupuncture,manual therapy,and Chinese materia medica;the main outcome indicators were scale scores;17 studies met the requirements for data sharing and management plans.Conclusion The number of registered TCM clinical trials for cervical spondylosis is generally increasing,involving multiple subtypes of cervical spondylosis and diversified intervention measures.But there are problems such as significant regional differences,uneven methodological quality,and poor data sharing,which should be improved.
5.Tauroursodeoxycholic acid treats spinal cord injury by reducing apoptosis of spinal cord neurons under glucose and oxygen deprivation
Zepeng CHEN ; Yonghui HOU ; Shudong CHEN ; Yu HOU ; Dingkun LIN
Chinese Journal of Tissue Engineering Research 2024;28(4):528-534
BACKGROUND:Tauroursodeoxycholic acid is a hydrophilic bile acid derivative that has neuroprotective effects in a variety of neurological disease models.However,there are few reports on the effects of tauroursodeoxycholic acid on spinal cord injury. OBJECTIVE:To investigate the effect of tauroursodeoxycholic acid on apoptosis of spinal cord neurons under hypoglycemic and hypoxic conditions,as well as the effect on recovery of motor function in mice after spinal cord injury. METHODS:(1)In vitro experiment:Primary spinal cord neurons were isolated from C57 BL/6 mouse embryos at 13.5 days of gestation.After 72 hours of culture,the cells were divided into three groups.In the normal group,cells were cultured in Neurobasal complete medium that was incubated in a CO2 incubator(5%CO2 + 95%air)for 24 hours.In the oxyglucose-deprived group,sugar-free Neurobasal medium was added and incubated in a triple-gas incubator(94%N2+5%CO2+1%O2)for 12 hours,and then the medium was replaced with Neurobasal complete medium and incubated in a CO2 incubator for 12 hours.In the experimental group,the treatment procedure was approximately the same as that in the oxyglucose-deprived group,except that taurodeoxycholic acid was added along with the sugar-free Neurobasal medium.TUNEL staining was used to detect apoptosis,cell counting kit-8 assay was applied to detect cell activity,and immunofluorescence staining was performed to detect cellular β-microtubule protein expression.(2)Animal experiment:Sixty C57 BL/6 mice were randomly divided into sham-operated group,spinal cord injury group and experimental group,with 20 mice in each group.Animal models of T9-T10 spinal cord injury were established using Allen's percussion method in the spinal cord injury group and the experimental group.Starting from the 1st day after modeling,taurodeoxycholic acid solution was given by gavage in the experimental group and normal saline was given by gavage in the sham-operated and spinal cord injury groups once a day for 14 consecutive days.Spinal cord tissue repair was assessed using behavioral and histological methods. RESULTS AND CONCLUSION:In vitro experiment:TUNEL staining,cell counting kit-8 and immunofluorescence staining showed that compared with the normal group,the number of apoptotic cells was higher(P<0.01),while cell activity and β-microtubule protein expression were lower in the oxyglucose-deprived group(P<0.01);compared with the oxyglucose-deprived group,the number of apoptotic cells was lower(P<0.01),while cell activity and β-microtubule protein expression were higher in the experimental group(P<0.01).Animal experiment:The Basso-Beattie-Bresnahan scores in the open field test and hind limb footprint experiments showed that the mice in the experimental group had better recovery of walking and motor functions than those in the spinal cord injury group.Hematoxylin-eosin staining showed that significant deformities and cavities were observed at the site of spinal cord injury and the number of nerve cells was significantly reduced in the spinal cord injury group.Compared with the spinal cord injury group,the experimental group showed a significant reduction in the area of spinal cord injury,less spinal cord deformity,fewer cavities,and an increase in the number of nerve cells.Immunofluorescence staining showed that the number of neuronal nucleus-labeled neuronal cells in the spinal cord injury group was less than that in the sham-operated group(P<0.01),and the number of neuronal nucleus-labeled neuronal cells in the experimental group was higher than that in the spinal cord injury group(P<0.01).To conclude,tauroursodeoxycholic acid could effectively reduce glucose/oxygen deprivation-induced apoptosis of spinal cord neurons and axonal loss,and promote the recovery of motor function in mice with spinal cord injury.
6.Clinical study of arthroscopic treatment for patients with borderline developmental dysplasia of the hip
Weiming YANG ; Yang LV ; Peng YANG ; Cong LI ; Da GUO ; Dingkun LIN
Chinese Journal of Sports Medicine 2024;43(8):605-612
Objective To explore the clinical effect of hip arthroscopy in the treatment of borderline developmental dysplasia of the hip(BDDH)with acetabular labrum injury.Methods A total of 32 BDDH patients with acetabular labrum injury and undergoing hip arthroscopy between December 2019 and December 2021 were analyzed retrospectively.There were 8 males and 24 females,aged 39.41±16.27 years(18~59 years)and their mean lateral acetabular central-edge angle(LCEA)was 21.25±2.96°(18°~25°).Another 32 patients with acetabular femur impingement(FAI)matching in age and undergoing hip arthroscopy during the same period were selected into the control group,which includ-ed 15 males and 17 females,aged 42.78±12.18 years(24~70 years),with their average LCEA of 32.97°±4.96°(26°~42°).Both groups received the arthroscopic glenolabial repair,head and neck ar-ea plasty and joint capsule suture.Three,six and twenty-four hours after the surgery,both groups were evaluated the clinical effect using the modified Harris hip score(mHHS),hip outcome score sports specific subscale(HOS-SSS),hip outcome score-activities of daily living(HOS-ADL)and visual analogue scale(VAS).Results The BDDH and FAI groups were followed up for(27.6±5.4)and(28.3±6.5)months,respectively.There was no significant difference in all measurements between the two groups before the operation(P>0.05).However,3,12 and 24 months after surgery,the aver-age mHHS,HOS-SSS and HOS-ADL scores of both groups increased significantly,while the average VAS scores decreased significantly(P<0.001).Moreover,the average mHHS scores of BDDH group were significantly lower than FAI group 3,12 and 24 months after surgery(P<0.05).However,3 months after surgery,the average HOS-SSS and HOS-ADL scores in BDDH group were significantly lower than FAI group(P<0.05).None of the patients underwent re-surgery due to recurrence of symp-toms during the follow-up period.Conclusion Hip arthroscopy can achieve satisfying short-term out-comes in treating BDDH.
7.Phyllanthi Fructus: A modal medicinal and food homologous item in quality evaluation.
Gefei LI ; Yurou JIANG ; Dingkun ZHANG ; Li HAN ; Taigang MO ; Sanhu FAN ; Haozhou HUANG ; Junzhi LIN
Chinese Herbal Medicines 2023;15(3):360-368
Phyllanthi Fructus is a highly unique medicine and food homologous item, which exhibits distinctive flavor, notable nutritional value, and abundant pharmacological activity. It has enormous potential in the creation of health products and pharmaceuticals. However, due to the unique laws of quality formation and transfer of Phyllanthi Fructus, its appearance, shape, chemical compositions, nutrients, and sensory flavors are frequently greatly influenced by botanical resources, the processing and storage conditions. As a result, the current quality evaluation model is difficult to meet the needs of Phyllanthi Fructus as a medicine and food homologous item in the development of diversified products. This paper constructs the hierarchical utilization mode of Phyllanthi Fructus based on its unique quality formation and transmission laws, explores the quality evaluation model for food-oriented use and medicinal-oriented use, respectively, and systematically describes the quality evaluation idea under diversified application scenarios. This paper aims to serve as a reference for the construction of a quality evaluation model suitable for the medicine and food homologous item of Phyllanthi Fructus.
8.A novel quantified bitterness evaluation model for traditional Chinese herbs based on an animal ethology principle.
Xue HAN ; Hong JIANG ; Li HAN ; Xi XIONG ; Yanan HE ; Chaomei FU ; Runchun XU ; Dingkun ZHANG ; Junzhi LIN ; Ming YANG
Acta Pharmaceutica Sinica B 2018;8(2):209-217
Traditional Chinese herbs (TCH) are currently gaining attention in disease prevention and health care plans. However, their general bitter taste hinders their use. Despite the development of a variety of taste evaluation methods, it is still a major challenge to establish a quantitative detection technique that is objective, authentic and sensitive. Based on the two-bottle preference test (TBP), we proposed a novel quantitative strategy using a standardized animal test and a unified quantitative benchmark. To reduce the difference of results, the methodology of TBP was optimized. The relationship between the concentration of quinine and animal preference index (PI) was obtained. Then the PI of TCH was measured through TBP, and bitterness results were converted into a unified numerical system using the relationship of concentration and PI. To verify the authenticity and sensitivity of quantified results, human sensory testing and electronic tongue testing were applied. The quantified results showed a good discrimination ability. For example, the bitterness of Coptidis Rhizoma was equal to 0.0579 mg/mL quinine, and Nelumbinis Folium was equal to 0.0001 mg/mL. The validation results proved that the new assessment method for TCH was objective and reliable. In conclusion, this study provides an option for the quantification of bitterness and the evaluation of taste masking effects.
9.Investigation of possible relationship between three-dimensional radiological localization of herniated lumbar discs and their functional status
Weiming YANG ; Feipeng NING ; Dingkun LIN
Chinese Journal of Orthopaedics 2014;34(3):280-284
Objective This study was designed to describe the localization and the extent of lumbar disc herniations (LDH) by three-dimensional radiological classification and to investigate their potential correlation with symptom and function evaluation.Methods The study retrospectively analyzed 261 patients (284 segments of herniated discs) who were diagnosed with LDH by computerized tomography (CT) or magnetic resonance imaging (MRI) from January 2006 to April 2010.There were 162 males and 99 females,with an average age of 42.1 years old (range,14-67 years) and the mean duration of history is 14.6 months (range,1-38 months).There were 35 cases of heavy manual workers,133 cases of moderate manual workers,and 93 cases of comfortable manual workers.The data were consisted of 188 MRI and 73 CT reports.The three-dimensional radiological classification was applied to describe the localization and the extent of LDH.Oswestry disability index (ODI) scores,Japanese Orthopaedic Association (JOA) scores and visual analogue scores (VAS) were applied to evaluate the vertebral symptom and function.The relationship between three-dimensional radiological classification and their functional status was probed.Results Herniations of lumbar discs located mainly sagittally at Level Ⅰ (151 segments,53.2%) and Level Ⅲ(122 segments,43.0%); horizontally at Area 2 (209 segments,73.6%)and Area 3 (61 segments,21.5%); and frontally at Area b (162 segments,57.0%),Area a (78 segments,27.5%) and Area c (41 segments,14.4%).The ODI scores at visit were 56.91%±13.62%,ranging from 20% to 90%;JOA scores were ranging from 0 to 24 (mean,13.57±4.68); while VAS scores were ranging from 3 to 10 (mean,6.09±1.89).There was no significant difference on ODI scores,JOA scores and VAS scores between different herniation locations.Conclusion According to the three-dimensional radiological classification,most of the lumbar disc herniations located at Level Ⅰ sagittally,Area 2 horizontally,and Area b frontally.Clinical symptoms and lumbar function did not depend on the location of the lumbar disc herniations by the three-dimensional classification.
10.Powder modification technology used for the preparation of the hydrophilic decoction pieces of indigo naturalis and the modification principle.
Dingkun ZHANG ; Junzhi LIN ; Jianyun LIU ; Chunfeng QIN ; Zhiping GUO ; Li HAN ; Ming YANG
Acta Pharmaceutica Sinica 2013;48(7):1148-55
The hydrophilicity of the normal decoction pieces (NDP) of Indigo Naturalis is not good, therefore, it is not suit for decoctions. In this paper, powder modification technology is used and some NDP and alcohol are ground together in the vibromill to prepare the hydrophilic decoction pieces (HDP) of Indigo Naturalis. Initially, the properties of NDP, ultrafine decoction pieces (UDP) and HDP are compared, the hydrophilicity of UDP was promoted slightly, that of HDP is promoted dramatically. Then, three batches of Indigo Naturalis are prepared to HDP separately, but there is no obvious difference in the contact angle. Furthermore, the size distribution, surface area and micro-shape of HDP are bigger than that of UDP and smaller than NDP. The contents of indigo and indirubin in three decoction pieces are the same, as well as the species of inorganic substance, although there is a little difference in the proportion of five inorganic substances. The fact suggests the change of physical state and the qualitative and quantitative change of organism and inorganic substances are not the main factors to influence the hydrophilicity. In addition, hydroxyl, methylene and methyl can be identified at the wavenumber of 3 356 cm(-1) and 1 461 cm(-1) in infrared spectrum; the content of alcohol in HDP is 0.67% measured by gas chromatogram. The stability of HDP in the heating condition is studied, the fact suggests the hydrophilic effect of HDP at 40 degrees C is relatively stable. All above research suggests that the alcohol is the main factor to influence the hydrophilicity and maybe the intermolecular force which fixed alcohol molecule on the surface of Indigo Naturalis is the basic principle to produce the hydrophilicity.

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