1.Identification of Original Plants of Lycii Cortex by Multiplex Allele-specific PCR
Yang SHI ; Li HU ; Yuyang ZHAO ; Chao JIANG ; Yan JIN ; Jing MU ; Yuan YUAN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(4):35-41
ObjectiveTo identify Lycium chinense and L. barbarum as the original plants of Lycii Cortex simply and efficiently by multiple allele-specific polymerase chain reaction (PCR). MethodThe chloroplast genome sequences of L. chinense and L. barbarum were downloaded from the Chloroplast Genome Information Resource (CGIR), and then IdenDSS was employed to screen out the specific single nucleotide polymorphism (SNP) sites between the two plants. Primer 5.0 was used to design the specific primers, including primers GQ-F/R for identifying L. chinense and primers NX-F/R for identifying L. barbarum. Furthermore, the primer concentration ratio, annealing temperature, cycles, and Taq enzyme were optimized to establish the optimal PCR system and conditions for plant identification. Finally, the applicability of the established method was examined with the plant samples collected from different regions. ResultThe PCR with GQ-F/R∶NX-F/R concentration ratio of 2∶1 at the annealing temperature at 59 ℃ and for 30 cycles showed specific bands at 183 bp and 295 bp, respectively, for L. chinense and L. barbarum samples from different regions. ConclusionThe established PCR approach can simply, rapidly, and efficiently identify the original plants of Lycii Cortex, serving as a new method for the discrimination between L. chinense and L. barbarum. Moreover, the method provides technical support for the research and development of classic famous prescriptions containing Lycii Cortex.
2.Relationship between lumbar spine development and hip development in children with spastic cerebral palsy
Xinwei ZUO ; Gang LIU ; Huizhong BAI ; Lin XU ; Yi ZHAO ; Jingpei REN ; Chuanyu HU ; Xiaohong MU
Chinese Journal of Tissue Engineering Research 2024;28(8):1247-1252
BACKGROUND:Most scholars now believe that children with cerebral palsy who have severe spinal deformities in early childhood(<15 years of age)may have a higher risk of progression of spinal deformities,which may result from imbalances in movement due to pelvic tilt,pain,etc. OBJECTIVE:To investigate the relationship between lumbar spine development and hip joint development in children with spastic cerebral palsy. METHODS:A retrospective analysis was performed in 102 children with spastic cerebral palsy admitted at Dongzhimen Hospital Affiliated to Beijing University of Chinese Medicine from January 2014 to December 2021.All admitted children had X-rays of the pelvic position and the lumbar lateral position.Anteroposterior X-ray of the pelvis was performed to measure femoral head migration percentage,central edge angle,neck-shaft angle,and acetabular index.The sagittal Cobb angle,sacral slope,arch-top distance,and lumbar lordosis index were measured by the lateral X-ray of the lumbar spine.Correlation of the two sets of indicators was further analyzed.All children were divided into normal group,risk group,hip subluxation group and total hip dislocation group according to their femoral head migration percentage,and the differences in lumbar spine indexes between groups were evaluated. RESULTS AND CONCLUSION:Pearson correlation analysis showed that the femoral head migration percentage was moderately positively correlated with sagittal Cobb angle and arch-top distance,and weakly positively correlated with lumbar lordosis index;the central edge angle was moderately negatively correlated with the arch-top distance and weakly negatively correlated with the sagittal Cobb angle;the neck-shaft angle was weakly positively correlated or not correlated with the sagittal Cobb angle and lumbar lordosis index;and the acetabular index was weakly positively correlated with the sagittal Cobb angle and arch-top distance.No statistically significant correlation was found between the remaining indicators.According to the femoral head migration percentage,the children were divided into four groups,including 25 cases in the normal group,41 cases in the risk group,27 cases in the hip subluxation group,and 9 cases in the total hip dislocation group.The sagittal Cobb angle was significantly increased in the risk group,the hip subluxation group and the total hip dislocation group compared with the normal group,showing an increasing trend group by group,and there were significant differences between groups(P<0.05).Compared with the normal group,the lumbar lordosis index in the risk group and the hip subluxation group increased significantly,and there were significant differences between groups(P<0.05).There was an increase trend in the lumbar lordosis index of the total hip dislocation group compared with the normal group.Compared with the normal group,the arch-top distance in the hip subluxation group and the total hip dislocation group increased significantly(P<0.05),and there was a stepwise increasing trend.There was no significant difference in sacral slope between groups.To conclude,the development of the lumbar spine in children with cerebral palsy is closely related to the development of the pelvic hip joint,and the most obvious relationship is between lumbar lordosis and hip dislocation.
3.The Role of α7nAChR in Alzheimer’s Disease
Dao-Bo DING ; Wen-Jun MU ; Xin LI ; Huan CHEN ; Hong-Wei HOU ; Qing-Yuan HU
Progress in Biochemistry and Biophysics 2024;51(11):2897-2904
As the global population continues to age, the incidence of Alzheimer’s disease (AD), one of the most common neurodegenerative diseases, continues to rise significantly. As the disease progresses, the patient’s daily living abilities gradually decline, potentially leading to a complete loss of self-care abilities. According to estimates by the Alzheimer’s Association and the World Health Organization, AD accounts for 60%-70% of all other dementia cases, affecting over 55 million people worldwide. The case number is estimated to double by 2050. Despite extensive research, the precise etiology and pathogenesis of AD remain elusive. Researchers have a profound understanding of the disease’s pathological hallmarks, which include amyloid plaques and neurofibrillary tangles resulting from the abnormal phosphorylation of Tau protein. However, the exact causes and mechanisms of the disease are still not fully understood, leaving a vital gap in our knowledge and understanding of this debilitating disease. A crucial player that has recently emerged in the field of AD research is the α7 nicotinic acetylcholine receptor (α7nAChR). α7nAChR is composed of five identical α7 subunits that form a homopentamer. This receptor is a significant subtype of acetylcholine receptor in the central nervous system and is widely distributed in various regions of the brain. It is particularly prevalent in the hippocampus and cortical areas, which are regions associated with learning and memory. α7nAChR plays a pivotal role in several neurological processes, including neurotransmitter release, neuronal plasticity, cell signal transduction, and inflammatory response, suggesting its potential involvement in numerous neurodegenerative diseases, including AD. In recent years, the role of α7nAChR in AD has been the focus of extensive research. Emerging evidence suggests that α7nAChR is involved in several critical steps in the disease progression of AD. These include involvement in the metabolism of amyloid β-protein (Aβ), the phosphorylation of Tau protein, neuroinflammatory response, and oxidative stress. Each of these processes contributes to the development and progression of AD, and the involvement of α7nAChR in these processes suggests that it may play a crucial role in the disease’s pathogenesis. The potential significance of α7nAChR in AD is further reinforced by the observation that alterations in its function or expression can have significant effects on cognitive abilities. These findings suggest that α7nAChR could be a promising target for therapeutic intervention in AD. At present, the results of drug clinical studies targeting α7nAChR show that these compounds have improvement and therapeutic effects in AD patients, but they have not reached the degree of being widely used in clinical practice, and their drug development still faces many challenges. Therefore, more research is needed to fully understand its role and to develop effective treatments based on this understanding. This review aims to summarize the current understanding of the association between α7nAChR and AD pathogenesis. We provide an overview of the latest research developments and insights, and highlight potential avenues for future research. As we deepen our understanding of the role of α7nAChR in AD, it is hoped that this will pave the way for the development of novel therapeutic strategies for this devastating disease. By targeting α7nAChR, we may be able to develop more effective treatments for AD, ultimately improving the quality of life for patients and their families.
4.Treatment of Insomnia by the Method of Unblocking the Bowels and Harmonizing Viscera Based on the Theory of "Viscera-Bowels Extraordinary Connection"
Bin WU ; Zhenyun HAN ; Wenyue HU ; Lei MU ; Biying ZHONG ; Hongpei LI
Journal of Traditional Chinese Medicine 2024;65(17):1769-1774
This paper aimed to explain the clinical thinking of using the method of unblocking the bowels and harmonizing viscera. Based on theory of "viscera-bowels extraordinary connection" of The Gateway to Medicine (《医学入门》), combined with the theoretical research, clinical practice and individual experience of ancient physicians, modern scholars, it is believed that viscera qi disorder caused by bowels qi blocking is the basic mechanism of insomnia. In clinic, it is common that gallbladder phlegm-heat harassing the heart spirit, stagnant blood and qi in stomach blocking the pericardium, turbid heat in large intestine causing fire and affecting liver, dampness insmall intestine trapping the spleen, water retention in the bladder drying up lungs, and the inhibited original qi of sanjiao damaging the kidney, which could be treated with Wendan Decoction (温胆汤), Taohe Chengqi Decoction (桃核承气汤), Chengqi-series Decoction (承气汤类方), Linggui Zhugan Decoction (苓桂术甘汤), Wuling Powder (五苓散), Xiaochaihu Decoction (小柴胡汤), respectively.
5.Comparison of Direct and Extraction Immunoassay Methods With Liquid Chromatography-Tandem Mass Spectrometry Measurement of Urinary Free Cortisol for the Diagnosis of Cushing’s Syndrome
Danni MU ; Jiadan FANG ; Songlin YU ; Yichen MA ; Jin CHENG ; Yingying HU ; Ailing SONG ; Fang ZHAO ; Qi ZHANG ; Zhihong QI ; Kui ZHANG ; Liangyu XIA ; Ling QIU ; Huijuan ZHU ; Xinqi CHENG
Annals of Laboratory Medicine 2024;44(1):29-37
Background:
Twenty-four-hour urinary free cortisol (UFC) measurement is the initial diagnostic test for Cushing’s syndrome (CS). We compared UFC determination by both direct and extraction immunoassays using Abbott Architect, Siemens Atellica Solution, and Beckman DxI800 with liquid chromatography-tandem mass spectrometry (LC-MS/MS). In addition, we evaluated the value of 24-hr UFC measured by six methods for diagnosing CS.
Methods:
Residual 24-hr urine samples of 94 CS and 246 non-CS patients were collected.A laboratory-developed LC-MS/MS method was used as reference. UFC was measured by direct assays (D) using Abbott, Siemens, and Beckman platforms and by extraction assays (E) using Siemens and Beckman platforms. Method was compared using Passing–Bablok regression and Bland–Altman plot analyses. Cut-off values for the six assays and corresponding sensitivities and specificities were calculated by ROC analysis.
Results:
Abbott-D, Beckman-E, Siemens-E, and Siemens-D showed strong correlations with LC-MS/MS (Spearman coefficient r = 0.965, 0.922, 0.922, and 0.897, respectively), while Beckman-D showed weaker correlation (r = 0.755). All immunoassays showed proportionally positive bias. The areas under the curve were 0.975 for Abbott-D, 0.972 for LCMS/MS, 0.966 for Siemens-E, 0.948 for Siemens-D, 0.955 for Beckman-E, and 0.877 for Beckman-D. The cut-off values varied significantly (154.8–1,321.5 nmol/24 hrs). Assay sensitivity and specificity ranged from 76.1% to 93.2% and from 93.0% to 97.1%, respectively.
Conclusions
Commercially available immunoassays for measuring UFC show different levels of analytical consistency compared to LC-MS/MS. Abbott-D, Siemens-E, and Beckman-E have high diagnostic accuracy for CS.
6.Stability study of umbilical cord mesenchymal stem cells formulation in large-scale production
Wang-long CHU ; Tong-jing LI ; Yan SHANGGUAN ; Fang-tao HE ; Jian-fu WU ; Xiu-ping ZENG ; Tao GUO ; Qing-fang WANG ; Fen ZHANG ; Zhen-zhong ZHONG ; Xiao LIANG ; Jun-yuan HU ; Mu-yun LIU
Acta Pharmaceutica Sinica 2024;59(3):743-750
Umbilical cord mesenchymal stem cells (UC-MSCs) have been widely used in regenerative medicine, but there is limited research on the stability of UC-MSCs formulation during production. This study aims to assess the stability of the cell stock solution and intermediate product throughout the production process, as well as the final product following reconstitution, in order to offer guidance for the manufacturing process and serve as a reference for formulation reconstitution methods. Three batches of cell formulation were produced and stored under low temperature (2-8 ℃) and room temperature (20-26 ℃) during cell stock solution and intermediate product stages. The storage time intervals for cell stock solution were 0, 2, 4, and 6 h, while for intermediate products, the intervals were 0, 1, 2, and 3 h. The evaluation items included visual inspection, viable cell concentration, cell viability, cell surface markers, lymphocyte proliferation inhibition rate, and sterility. Additionally, dilution and culture stability studies were performed after reconstitution of the cell product. The reconstitution diluents included 0.9% sodium chloride injection, 0.9% sodium chloride injection + 1% human serum albumin, and 0.9% sodium chloride injection + 2% human serum albumin, with dilution ratios of 10-fold and 40-fold. The storage time intervals after dilution were 0, 1, 2, 3, and 4 h. The reconstitution culture media included DMEM medium, DMEM + 2% platelet lysate, 0.9% sodium chloride injection, and 0.9% sodium chloride injection + 1% human serum albumin, and the culture duration was 24 h. The evaluation items were viable cell concentration and cell viability. The results showed that the cell stock solution remained stable for up to 6 h under both low temperature (2-8 ℃) and room temperature (20-26 ℃) conditions, while the intermediate product remained stable for up to 3 h under the same conditions. After formulation reconstitution, using sodium chloride injection diluted with 1% or 2% human serum albumin maintained a viability of over 80% within 4 h. It was observed that different dilution factors had an impact on cell viability. After formulation reconstitution, cultivation in medium with 2% platelet lysate resulted in a cell viability of over 80% after 24 h. In conclusion, the stability of cell stock solution within 6 h and intermediate product within 3 h meets the requirements. The addition of 1% or 2% human serum albumin in the reconstitution diluent can better protect the post-reconstitution cell viability.
7.Rapid Screening of 34 Emerging Contaminants in Surface Water by UHPLC-Q-TOF-MS
Chen-Shan LÜ ; Yi-Xuan CAO ; Xiao-Xi MU ; Hai-Yan CUI ; Tao WANG ; Zhi-Wen WEI ; Ke-Ming YUN ; Meng HU
Journal of Forensic Medicine 2024;40(1):30-36
Objective To establish a rapid screening method for 34 emerging contaminants in surface water by ultra-high performance liquid chromatography-quadrupole-time of flight mass spectrometry(UHPLC-Q-TOF-MS).Methods The pretreatment conditions of solid phase extraction(SPE)were op-timized by orthogonal experimental design and the surface water samples were concentrated and ex-tracted by Oasis? HLB and Oasis? MCX SPE columns in series.The extracts were separated by Kine-tex? EVO C18 column,with gradient elution of 0.1%formic acid aqueous solution and 0.1%formic acid methanol solution.Q-TOF-MS'fullscan'and'targeted MS/MS'modes were used to detect 34 emerging contaminants and to establish a database with 34 emerging contaminants precursor ion,prod-uct ion and retention times.Results The 34 emerging contaminants exhibited good linearity in the con-centration range respectively and the correlation coefficients(r)were higher than 0.97.The limit of de-tection was 0.2-10 ng/L and the recoveries were 81.2%-119.2%.The intra-day precision was 0.78%-18.70%.The method was applied to analyze multiple surface water samples and 6 emerging contaminants were detected,with a concentration range of 1.93-157.71 ng/L.Conclusion The method is simple and rapid for screening various emerging contaminants at the trace level in surface water.
8.Correlation between preoperative plasma fibrin degradation products level and clinicopathological features in patients with non-small cell lung cancer
Juanjuan CHE ; Jing WANG ; Mu HU ; Hongchao ZHEN ; Haishan LIN ; Kun SHANG ; Bangwei CAO
Cancer Research and Clinic 2024;36(1):1-5
Objective:To investigate the relationship between preoperative plasma fibrin degradation products (FDP) level and clinicopathological features of patients with completely resected non-small cell lung cancer (NSCLC).Methods:A retrospective case series study was performed. The clinical data of 521 patients who were pathologically diagnosed with NSCLC in Beijing Friendship Hospital Affiliated to Capital Medical University from January 2016 to December 2017 were retrospectively analyzed. Among 521 cases, 406 cases were postoperatively pathologically confirmed as non-lymph node and non-distant metastasis (non-metastasis group) and 115 cases were postoperatively pathologically confirmed as lymph node or distant metastasis (metastasis group). The preoperative FDP level and clinicopathological characteristics as well as the clinicopathological characteristics of NSCLC patients with different FDP levels were compared between the two groups. The correlation between preoperative FDP level and TNM staging was analyzed by using Spearman correlation analysis.Results:Among 521 NSCLC patients, 266 cases were female, 255 cases were male; the age [ M( Q1, Q3)] was 59 years (54 years, 65 years); 441 cases were adenocarcinoma and 70 cases were squamous cell carcinoma. The preoperative median FDP level was 2.78 mg/L (2.35 mg/L, 3.13 mg/L) and 2.99 mg/L (2.56 mg/L, 4.16 mg/L), respectively of NSCLC patients in non-metastasis group and metastasis group, and the difference was statistically significant ( Z = 6.13, P < 0.001). The preoperative FDP level was 2.56 mg/L (2.35 mg/L, 3.20 mg/L) and 2.99 mg/L (2.56 mg/L, 3.20 mg/L), respectively in the early-stage NSCLC (stage Ⅰ-Ⅱ) and advanced NSCLC (stage Ⅲ-Ⅳ) patients, and the difference was statistically significant ( Z = 8.42, P < 0.001). Spearman correlation analysis showed that preoperative FDP level was positively correlated with tumor diameter ( r = 0.287, P < 0.001). There was a positive correlation between preoperative FDP level and the number of metastatic lymph nodes in 115 patients with lymph node metastasis ( r = 0.679, P < 0.001). According to the preoperative median FDP (2.78 mg/L), all patients were divided into FDP ≤2.78 mg/L group and FDP >2.78 mg/L, and there were statistically significant differences in age, metastasis, tumor staging, tumor diameter, the metastatic number of lymph node and histological types of NSCLC patients in both groups (all P < 0.05). Conclusions:The increase of preoperative plasma FDP level may be related to the tumor metastasis and clinical stage of NSCLC patients
9.Associations of genetic variants in GLP-1R with blood pressure responses to dietary sodium and potassium interventions
Mingke CHANG ; Chao CHU ; Mingfei DU ; Hao JIA ; Yue SUN ; Guilin HU ; Xi ZHANG ; Dan WANG ; Wenjing LUO ; Yu YAN ; Ziyue MAN ; Yang WANG ; Jianjun MU
Journal of Xi'an Jiaotong University(Medical Sciences) 2024;45(2):212-218
【Objective】 To investigate the association between genetic variations in the glucagon-like peptide-1 receptor (GLP-1R) gene and BP responses to sodium and potassium intake. 【Methods】 A total of 514 subjects from 124 families were recruited in Meixian County, Shaanxi Province, in 2004, resulting in the establishment of a "salt-sensitive hypertension study cohort" . The subjects followed a dietary regimen which involved a normal diet for 3 days, a low-salt diet for 7 days, a high-salt diet for 7 days, and a high-salt potassium-supplemented diet for 7 days. BP measurement was conducted at different intervention periods, and peripheral blood samples were collected. Additionally, eight single nucleotide polymorphisms (SNPs) of the GLP-1R gene were genotyped using the MassARRAY detection platform. 【Results】 The GLP-1R gene SNP rs9462472 exhibited a significant association with systolic BP, diastolic BP, and mean arterial pressure response to high-salt intervention. Similarly, SNP rs2268637 showed a significant association with systolic BP response to high-salt intervention. Furthermore, SNP rs2268637 was significantly associated with systolic BP and mean arterial pressure responses to high-salt plus potassium supplementation intervention. 【Conclusion】 Our findings indicate a significant association of genetic variations in the GLP-1R gene with BP responses to sodium and potassium intake. This suggests that the GLP-1R gene plays a role in the regulation of BP salt sensitivity and potassium sensitivity.
10.Shujin Jiannao Prescription alleviates inflammation in the cerebral cortex of rats with hypoxic-ischemic cerebral palsy
Gang LIU ; Jie ZENG ; Yalin ZHAO ; Bowen DENG ; Shengyuan JIANG ; Yaqi ZHANG ; Yi ZHAO ; Jingpei REN ; Chuanyu HU ; Lin XU ; Xiaohong MU
Chinese Journal of Tissue Engineering Research 2024;28(23):3674-3679
BACKGROUND:Shujin Jiannao Prescription is an empirical formula for the treatment of cerebral palsy in Dongzhimen Hospital,Beijing University of Chinese Medicine,with clear clinical efficacy,but the specific mechanism needs to be elucidated. OBJECTIVE:To explore the possible mechanism of Shujin Jiannao Prescription in treating cerebral palsy. METHODS:Sixty-four 7-day-old Sprague-Dawley rats were randomly divided into a normal group(n=12)and a model group(n=52).An animal model was established by the Rice-Vannucci method.After successful modeling,52 model rats were randomly divided into control model group(n=12),minocycline group,and the low-,medium-,and high-dose groups of the Shujin Jiannao Prescription(n=10 per group).Rats in the minocycline group were given 40 mg/kg·d minocycline by gavage;rats in the low-,medium,and high-dose groups were given 4,8,and 16 g/kg·d Shujin Jiannao Prescription granules by gavage,respectively;and rats in the normal group and control model group were given an equal dose of normal saline by gavage.Medication in each group was given once a day for 1 week.The rats in each group were evaluated behaviorally using suspension test,abnormal involuntary movement score,and Bederson score.The pathological changes of the cerebral cortex were observed by hematoxylin-eosin staining.The levels of tumor necrosis factor α,interleukin 1β,and interleukin 10 in the cerebral cortex were determined using ELISA.The positive expressions of Janus kinase 2(JAK2),phosphorylated Janus kinase 2(p-JAK2),phosphorylated signal transducer and activator of transcription 3(p-STAT3)in the cerebral cortex were detected using immunohistochemistry.The protein expression levels of JAK2,p-JAK2,and p-STAT3 were detected using western blot. RESULTS AND CONCLUSION:Compared with the normal group,the suspension test score and involuntary movement score were decreased in the control model group(P<0.01 or P<0.05).The pathological results showed structural disruption of nerve cells,formation of large numbers of vacuoles,cell swelling,and increased intercellular space in the control model group.In addition,the expressions of tumor necrosis factor α and interleukin 1β in the cerebral cortex were significantly increased(P<0.01),the expression of interleukin 10 was decreased(P<0.05),and the protein expressions of JAK2,p-JAK2,and p-STAT3 in the cerebral cortex were significantly increased(P<0.01)in the control model group compared with the normal group.Compared with the model group,minocycline and Shujin Jiannao Prescription at each dose could improve the behavioral indexes of rats(P<0.01 or P<0.05)and ischemic-hypoxic pathological changes were attenuated,with only a small amount of necrotic nerve cells and a few vacuoles,and reduced intercellular space.Moreover,the expressions of tumor necrosis factor α and interleukin 1β in the cerebral cortex were decreased in each drug group compared with the control model group(P<0.05),while the protein expressions of JAK2,p-JAK2,and p-STAT3 in the cerebral cortex were significantly decreased(P<0.01).The most obvious improvement was observed in the high-dose Shujin Jiannao Prescription group.To conclude,Shujin Jiannao Prescription can inhibit inflammation in the cerebral cortex of rats with hypoxic-ischemic brain injury.The mechanism may be related to the regulation of the JAK2/STAT3 signaling pathway.

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