1.Macrophage polarization in calcium oxalate nephrolithiasis:unraveling its role and influencing factors
Weisheng LI ; Chuanguo ZHANG ; Yangdong LI ; Wenqiang HE
Chinese Journal of Pathophysiology 2024;40(1):180-186
The most prevalent kind of renal calculi,calcium oxalate(CaOx),is characterized by its propensi-ty for recurrence in the urinary system.The development of CaOx renal calculi is greatly affected by macrophage polariza-tion.Particular oxalate causes an imbalance in macrophage polarization,which skews the M1/M2 ratio and makes it easier for CaOx crystals to accumulate in the kidneys and grow into calcium plaques in the renal papilla.Notably,M2 macro-phages can prevent CaOx renal calculi by consuming crystals and reducing inflammatory stress.As a result,immunothera-peutic techniques that alter M1 and M2 macrophage polarization are extremely promising for preventing CaOx renal calcu-li.To clarify the respective roles of M1 and M2 macrophages in the formation of CaOx crystals and provide insights for de-veloping immunotherapeutic interventions against CaOx renal calculi,this review summarizes the mechanisms underlying macrophage polarization in the genesis of CaOx renal calculi.
2.Rehmanniae Radix Praeparata Improves Neurological Function of Ischemic Stroke Rats by Inhibiting Autophagy and Ferroptosis
Saifei LI ; Peipei YUAN ; Yaxin WEI ; Liyuan GAO ; Panying LI ; Yuan RUAN ; Yi CHEN ; Yang FU ; Xiaoke ZHENG ; Weisheng FENG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(8):26-33
ObjectiveTo investigate the effect of Rehmanniae Radix Praeparata on neurological function injury in ischemic stroke rats and explore its mechanism. MethodMale SD rats were randomized into sham operation, model, low- and high -dose (3.5 g·kg-1 and 7 g·kg-1) Rehmannia Radix Praeparata, and nimodipine (0.01 g·kg-1) groups. The rat model of middle cerebral artery occlusion (MCAO) was established with the modified suture occlusion method. Zea-Longa 5-point scoring was employed to evaluate the neurological function of rats. The cerebral infarction volume was detected by 2,3,5-triphenyltetrazolium chloride (TTC) staining. Hematoxylin-eosin staining and Nissl staining were employed to observe the morphology and damage of the brain tissue. Meanwhile, the serum levels of lactate dehydrogenase (LDH), oxidative stress-related indicators superoxide dismutase (SOD), glutathione peroxidase 4 (GPX4), and malondialdehyde (MDA), and the iron (Fe) content in the brain tissue were determined. To explore the mechanism of Rehmanniae Radix Preparata in mitigating the neurological damage in ischemic stroke rats, Western blotting was employed to determine the expression levels of proteins in the ischemic brain tissue. The autophagy-associated proteins included autophagy effector (beclin-1), microtubule-associated protein light chain 3 (LC3B), and ubiquitin-binding protein p62 (p62). The ferroptosis-associated proteins included transferrin (TF), transferrin receptor 1 (TFR1), ferritin heavy chain 1 (FTH1), and ferropotin (FPN1). The neurological function injury-associated proteins included brain-derived neurotrophic factor (BDNF) and tyrosine kinase receptor B (TrkB). ResultCompared with the sham operation group, the model group showed increased neurological function score, cerebral infarction volume, and appearance of nuclear pyknosis and vacuole of cells in the cerebral cortex. In addition, the model group presented elevated levels of LDH, MDA, and Fe (P<0.01) and lowered levels of SOD and GPX4 (P<0.01). Compared with the model group, Rehmanniae Radix Praeparata decreased the content of LDH, MDA, and Fe (P<0.05, P<0.01) and elevated the levels of SOD and GPX4 (P<0.05, P<0.01). Compared with the sham operation group, the modeling promoted the expression of beclin-1,LC3B Ⅱ/Ⅰ, TF, and TFR1 and inhibited the expression of p62, FTH1, FPN1, BDNF, and TrkB (P<0.01). The expression levels of these proteins were recovered after the treatment with Rehmanniae Radix Praeparata. ConclusionRehmanniae Radix Praeparata may inhibit ferroptosis and improve the neurological function in ischemic stroke rats by down-regulating the autophagy level in the brain tissue.
3.Analysis of the results of screening for carriers of monogenic genetic diseases in 604 couples of childbearing age
Qianyun LI ; Renhua WU ; Yuanyuan ZHANG ; Fang LIU ; Weisheng CHENG ; Jing YUAN
Acta Universitatis Medicinalis Anhui 2024;59(9):1653-1658
Objective To understand the carrying situation and common variation of pathogenic genes of single gene hereditary disease in childbearing age population in Anhui province,to explore the establishment of clinical application network and referral model of carrier screening in Anhui province,and to explore the application value of expansible carrier screening(expanded carrier screening,ECS)in clinic.Methods Samples were collected from 604 individuals of childbearing age,all exhibiting a normal phenotype and a family history of inherited dis-ease.These samples were obtained during the first trimester or early stages of pregnancy(≤13+6 weeks).Based on high-throughput sequencing and special PCR analysis techniques,pathogenic variants associated with 220 disea-ses were detected,and related genes were detected in the spouses of positive carriers.Results As of May 16,2023,604 tested samples had been collected,and 340 carriers of the target disease had been detected;The posi-tive rate of pathogenic variation detection was 56.29% ;A total of 499 pathogenic variants were detected,with each tested individual carrying 0-5 variants;216 cases,accounting for 35.76% ,carried a single gene recessive dis-ease pathogenic variation,which was the most common.There were 95 cases carrying two types of single gene re-cessive genetic disease pathogenic variation,accounting for 15.73% .As of now,302 couples have been reported,and a total of 7 high-risk couples have been found through screening,with a high-risk rate of 2.32% .There are a total of 5 pairs with autosomal recessive genetic pattern(both spouses carry the same pathogenic gene),and 2 pairs with X-linked genetic pattern(the female carries the X-linked pathogenic gene).Conclusion In this study,we obtained the overall carrier and clinical application of target diseases as well as the carrier rates of causative genes of common single-gene genetic diseases in 604 subjects who underwent ECS testing,which could provide scientific guidance for the establishment of a clinical application network and referral model for carrier screening in Anhui Province.
4.Screening and genotyping of Mur blood group among voluntary blood donors in the population of Hezhou,Guangxi
Weiquan YUAN ; Shaohua DING ; Jianmin LI ; Xueming WU ; Shengming WEN ; Houquan LIN ; Weisheng HE ; Xi-Aoming LI ; Jiajie ZHANG ; Longming XIAO ; Shengbao DUAN ; Shengwang CHEN
Chinese Journal of Blood Transfusion 2024;37(7):773-778
Objective To screen the distribution frequency of Mur blood group among voluntary blood donors in Hezhou,Guangxi,and further analyze the molecular basis of of Mur antigen positive samples.Methods The Mur pheno-type of voluntary blood donors in Hezhou was serologically screened using microplate method,and the distribution frequency of Mur antigens in different ethnic groups was analyzed.Genetic typing was performed on these positive samples with PCR-SSP method to verify the accuracy of the serological method,and the genetic background was sequenced and analyzed.Re-sults Among 3 298 samples from voluntary blood donors in Hezhou,432(13.10%,432/3 298)were screened positive for Mur antigen,and PCR-SSP genotyping validation showed that all 432 samples were electrophoretic positive.Among them,the proportion of Han blood donors with positive Mur antigen was12.79%(331/2 587),Yao ethnic group was13.25%(64/483),Zhuang ethnic group was 16.51%(36/218),and no statistically significant difference was found in the three groups(P>0.05).Further sequencing results showed that 428 samples were GYP(B-A-B)Mur,also known as GYP.Mur type(12.98%,428/3 298),the other 4 samples were GYP(B-A-B)Bun,also known as GYP.Bun type(0.12%,4/3 298).Conclusion The Mur blood type frequency is high in the voluntary blood donors in Hezhou,Guangxi,and is predominant characterized by GYP.Mur genotype.Due to ethnic integration,no significant difference was noticed in the frequency of Mur blood type distribution between Han,Zhuang and Yao population.Therefore,conducting extensive Mur blood group antigen and antibody testing in Hezhou is of great significance for ensuring clinical blood transfusion safety.
5.Analysis of volatile constituents in different parts of Huai chrysanthemum by GC-MS combined with stoichiometry
Mengzhen GUO ; Meng LI ; Xiaoyan DENG ; Shuyan LIU ; Xiaolan WANG ; Xiaoya SUN ; Jingke ZHANG ; Xiaoke ZHENG ; Weisheng FENG
China Pharmacist 2024;27(2):209-219
Objective To analyze and identify the volatile constituents in different parts(flowers,stems and leaves)of Huai chrysanthemumin,and to lay a theoretical foundation for the comprehensive utilization for it.Methods The volatile oil in different parts of Huai chrysanthemumin were extracted by hydrodistillation,respectively.Their constituents were analyzed by gas chromatography-mass spectrometry(GC-MS).The compounds were identified by library search and literature screening.The relative percentage of each compound was obtained by the area normalization method.The differences in their chemical compositions were analyzed by Venn diagram,principal component analysis(PCA)and cluster heat map analysis.Results A total of 62 volatile chemical components were identified from different parts of Huai chrysanthemumin,including monoterpenes,sesquiterpenes,and their derivatives,as well as a small amount of aliphatic compounds.32,42 and 40 volatile components were detected from the flowers,stems and flowers,respectively.Furthermore 17 volatile components were shared by three parts,whereas 5,6 and 16 volatile components were unique to the flowers,stems and leaves,respectively.The results of stoichiometric analysis showed that both PCA and cluster heat map analysis could separate the flowers,stems and leaves,and their volatile components were different.Conclusion The types and contents of the volatile oil in the stems,leaves and flowers of Huai chrysanthemumin have certain variability,which provide a scientific basis for the further medicinal or industrial exploitation of different parts of Huai chrysanthemumin.
6.Retrospective analysis of 350 cases with dissection of lymph nodes posterior to right recurrent laryngeal nerve in endoscopic thyroidectomy through gasless axillary posterior approach
Zhicheng ZHANG ; Tingting LI ; Shitong YU ; Junna GE ; Zhigang WEI ; Baihui SUN ; Weisheng CHEN ; Jie TAN ; Shangtong LEI
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2024;59(1):21-26
Objective:To evaluated the safety and feasibility of dissection of lymph nodes posterior to right recurrent laryngeal nerve (ⅥB compartment) in endoscopic thyroidectomy through gasless axillary posterior approach.Methods:A total of 350 cases with right lobe papillary thyroid carcinoma (PTC) who underwent endoscopic lobectomy, isthmusectomy and central compartment neck dissection via gasless axillary posterior approach based at the Department of General Surgery, Nanfang Hospital, Southern Medical University from June 2020 to December 2022 were retrospectively analyzed. Summarize the clinical, pathological characteristics, and postoperative complications of the patients. SPSS 25.0 was used for statistical analysis of the data.Results:All 350 patients underwent endoscopic surgery successfully, with no conversion to open surgery. There were 303 females and 47 males, with an average age of (36.3±9.2) years. Of those, 287 patients were in pT1a stage, 62 in pT1b stage, and one patient in pT2 stage. There was no T3 or T4 stage patient. The mean numbers of yielded lymph nodes in right central compartment and ⅥB compartment were 8.11±4.65 (range, 1-31) and 2.62±1.86 (range, 1-12), respectively. ⅥB compartment metastasis was detected in 52 (14.86%) of 350 patients. The incidence of transient recurrent laryngeal nerve injury was 0.86%(3/350). Postoperative hematoma occurred in three patients (0.86%).Conclusion:The dissection of ⅥB compartment in endoscopic thyroidectomy through gasless axillary posterior approach is safe and feasible in selected PTC patients
7.Effect of Shilinqing granules on calcium oxalate nephrolithiasis in rats and its mechanism
Xiong YANG ; Xiaoxiao JIN ; Weisheng LI ; Cong ZHENG ; Wenqiang HE
China Pharmacy 2024;35(22):2750-2755
OBJECTIVE To explore the effect of Shilinqing granules on calcium oxalate nephrolithiasis in rats and its potential mechanisms through the silence information regulator 1 (SIRT1)/nuclear factor-κB (NF-κB)/nucleotide-binding domain leucine-rich repeat and pyrin domain-containing receptor 3 (NLRP3) signaling pathway. METHODS Sixty male SD rats were randomly assigned to control group, model group, and low- , medium- , high-dose groups of Shilinqing granules (6.5, 13, 26 g/kg, calculated based on crude drug), and high-dose of Shilinqing granules+inhibitor group (Shilinqing granules 26 g/kg+SIRT1 inhibitor nicotinamide 5 mg/kg), with 10 rats in each group. Except for the control group, the remaining groups of rats were given 1% ethylene glycol solution to drink freely and were intubated with 2% ammonium chloride solution 2 mL (once a day, for 4 weeks) to construct a calcium oxalate nephrolithiasis model. At the same time of modeling, the administration groups were intubated or (and) intraperitoneally injected with the corresponding drug solutions, while the control and the model groups were intubated with physiological saline and intraperitoneally injected with dimethyl sulfoxide. The body weight, kidney index, urine/ blood biochemical indicators [24-hour urine volume, urine pH, urinary calcium ion (Ca2+) and urinary oxalic acid (Ox) content, as well as blood creatinine (Scr), blood urea nitrogen (BUN), blood Ca2+ content], serum inflammatory indicators [levels of interleukin-1β (IL-1β), IL-18], pathological changes in renal tissue, calcium oxalate crystallization, and crystal scoring were observed. The protein expressions of SIRT1, NLRP3, and NF-κB in renal tissue were detected. RESULTS Compared with the control group, the model group had severe renal tissue damage and a large number of calcium oxalate crystals, with significant decrease or downregulation in body weight, 24-hour urine volume, urine pH, and protein expression of SIRT1 in renal tissue (P< 0.05); crystal score, kidney index, urinary contents of Ca2+ and Ox, serum contents of Scr, BUN and Ca2+, and serum levels of IL- 1β and IL-18, as well as the protein expressions of NF-κB, NLRP3 in renal tissue were significantly increased or upregulated (P<0.05). The pathological changes in the rats of 2024ZY2045) each dose group of Shilinqing granules were improved, the calcium oxalate crystals were reduced, and all quantitative indicators were significantly improved as compared with the model group (P<0.05); while the SIRT1 inhibitor could significantly reverse the improving effects of high-dose of Shilinqing granules on the above indicators (P<0.05). CONCLUSIONS Shilinqing granules can inhibit the formation of calcium oxalate nephrolithiasis in rats, reduce the levels of inflammatory indicators and its mechanism may be related to upregulating protein expression of SIRT1, and downregulating protein expressions of NF-κB and NLRP3.
8.Study on Quality Evaluation of Notopterygii Rhizoma et Radix through UPLC Fingerprint Combined with Chemometrics Analysis and Multi Index Component Content Determination
Guangming HE ; Xiaoying LU ; Tianrui XIA ; Feifei XIE ; Rui LUO ; Weisheng LYU ; Yueyi LIANG ; Zhenyu LI ; Xiuzhi LI ; Jianxiang ZHANG ; Xiangdong CHEN ; Dongmei SUN
Chinese Journal of Information on Traditional Chinese Medicine 2024;31(8):126-132
Objective To establish the ultra-high performance liquid chromatography(UPLC)chromatographic fingerprint of Notopterygii Rhizoma et Radix;To determine the contents of ferulic acid,nodakenin,ammijin,notopterol,isoimperatorin and volatile oil of Notopterygii Rhizoma et Radix from different producing areas;To provide reference for quality evaluation of Notopterygii Rhizoma et Radix.Methods Waters BEH C18 chromatographic column(2.1 mm×150 mm,1.7 μm)was used,with mobile phase acetonitrile-0.02%formic acid aqueous solution gradient elution,flow rate 0.25 mL/min,column temperature 25℃,detection wavelength 330 nm,injection volume 2 μL.UPLC fingerprints of 25 batches of Notopterygii Rhizoma et Radix were established,and the similarity analysis and chemometrics analysis were carried out.The contents of ferulic acid,nodakenin,ammijin,notopterol and isoimperatorin were determined simultaneously,and the contents of volatile oil was determined by steam distillation method.Results Totally 23 common fingerprint peaks were calibrated,11 known components were identified.According to the results of the cluster analysis and principal component analysis,25 batches of Notopterygii Rhizoma et Radix samples were divided into 3 categories,and the 6 potential differential components were screened out by orthogonal partial least squares-discriminant analysis(OPLS-DA).The results showed that the contents of notopterol and volatile oil from Sichuan Province were higher than those from Gansu Province and Qinghai Province.Conclusion The method established in the study is accurate and reliable,which can provide scientific basis and reference for the quality evaluation and control of Notopterygii Rhizoma et Radix.
9.Mechanisms of Qinghua Liangxue Huluo Decoction in preventing acute radiation enteritis in rats through the PI3K/Akt pathway
Lin ZHU ; Ting ZHANG ; Huafang YIN ; Weisheng SHEN ; Yu LI ; Jian WANG ; Weidong SHEN
Chinese Journal of Radiological Medicine and Protection 2024;44(7):571-577
Objective:To explore the regulatory effects of Qinghua Liangxue Huluo Decoction on oxidative stress and inflammation in acute radiation enteritis in rats, as well as its impact on the PI3K/Akt pathway. Methods:A total of 36 SD rats were randomly divided into four groups using block randomization, namely the control, model, low-dose group (6.17 g/kg), and high-dose (24.68 g/kg) groups, with nine rats in each group. These rats were exposed to X-ray irradiation at a dose of 17.5 Gy to induce acute radiation enteritis, followed by continuous intragastric administration for seven days pre- and post-irradiation. Seven days post-irradiation, the perianal and fecal conditions of rats in each group were observed, and rectal tissues were collected and ground. Enzyme-linked immunosorbent assay (ELISA) was employed to assess the activity of superoxide dismutase (SOD), catalase (CAT) expression, and malondialdehyde (MDA) levels indicative of lipid peroxidation. Quantitative real-time polymerase chain reaction (qRT-PCR) was utilized to analyze the mRNA expression of tumor necrosis factor-ɑ (TNF-ɑ), interleukin-6 (IL-6), and interleukin-1β (IL-1β) in the rectal tissues of each group. Additionally, Western blot was conducted to examine the expression of proteins associated with the PI3K/Akt signaling pathway in rectal tissues. The IEC-6 cells were categorized into the control, radiation, blank, and drug administration groups, with all these groups except for the control group subjected to 10 Gy single irradiation. ELISA was then employed to determine the concentrations of SOD, CAT, MDA, TNF-ɑ, IL-6, and IL-1β in cell supernatants, while Western blot was utilized to assess the expression of PI3K/Akt signaling pathway-related proteins in each group.Results:Compared to the model group, rats in the low-dose and high-dose groups exhibited a trend toward normal perianal and fecal conditions, increased SOD activity ( t = 4.86, 8.50, P < 0.05), elevated CAT expression ( t = 8.72, 14.28, P<0.05), and decreased MDA level ( t = 6.94, 10.66, P < 0.05). Furthermore, the mRNA expression of TNF-ɑ, IL-6, and IL-1β in rectal tissues was significantly inhibited in both low-dose and high-dose groups ( t = 5.60, 2.95, 4.31, 9.16, 4.66, 13.35, P < 0.05), along with lower p-PI3K/PI3K and p-Akt/Akt ratios in rectal tissues compared to the model group ( t = 22.35, 13.56, 18.23, 13.85, P < 0.05). Compared to the radiation group, the drug administration groups (10% drug-containing serum) exhibited increased SOD and CAT expressions ( t = 6.85, 10.44, P < 0.05), as well as decreased MDA expression ( t = 10.44, P < 0.05), in the supernatant. Furthermore, compared to the radiation group, this group displayed significantly inhibited TNF-ɑ, IL-6, and IL-1β concentrations in the cell supernatant ( t = 12.07, 6.87, 14.80, P < 0.05), while lowering p-PI3K/PI3K and p-Akt/Akt ratios in cells ( t = 10.95, 5.59, P < 0.05). Conclusions:Qinghua Liangxue Huluo Decoction demonstrates the potential for mitigating oxidative stress-induced injury and suppressing the expressions of inflammatory factors in rats with acute radiation enteritis. The mechanism behind the potential is likely associated with the negative regulation of the PI3K/Akt signaling pathway.
10.Establishment of UPLC characteristic chromatogram of Pulsatilla chinensis and its application in origin differentiation and counterfeit identification
Guangming HE ; Rui LUO ; Heping ZENG ; Xiaoying LU ; Xiaolong YANG ; Weisheng LYU ; Yueyi LIANG ; Zhenyu LI ; Dongmei SUN ; Xiangdong CHEN
International Journal of Traditional Chinese Medicine 2024;46(6):743-749
Objective:To establish ultra performance liquid chromatography (UPLC) characteristic chromatogram of Pulsatilla chinensis; To provide reference for the origin identification and quality control of Pulsatilla chinensis. Methods:UPLC Method was adopted. The determination was performed on a column of Agilent SB C18 (2.1 mm×100 mm, 1.8 μm) . The mobile phase was acetonitrile-methanol (2:1) -0.1% phosphoric acid solution by fradient elution at a flow rate of 0.30ml/min. The column temperature was 30 ℃. The detection wavelength was 215 nm. The injection volume was 2 μl. The common counterfeit products and medicinal herbs of Pulsatilla chinensis from different areas were evaluated by comparison of characteristic chromatogram, principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA). Results:There were 9 characteristic peaks in the characteristic chromatogram of Pulsatilla chinensis, and 8 common peaks were identified by high resolution mass spectrometry and comparison of reference materials. Through PCA analysis, it was possible to clearly distinguish the medicinal herbs of Pulsatilla chinensis from different areas. Combined with OPLS-DA analysis, it was found that peak 2, peak 3, peak 6 were the main markers of Pulsatilla chinensis from different producing areas. Conclusion:The established method has good specificity, repeatability and durability, and it can effectively distinguish the common counterfeits of Pulsatilla chinensis, and provide the basis of quality control and selection of origin for Pulsatilla chinensis.


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