1.Genome-wide association study of rubella virus vaccine strain BRD-Ⅱ
Yingmei XU ; Yongqiang ZHU ; Xin ZHOU ; Zhaoyang LIU ; Leijun MA ; Zhewen CHEN ; Yueye ZHAO ; Tiaoxia ZHU
Shanghai Journal of Preventive Medicine 2025;37(5):461-466
ObjectiveTo perform a genome-wide association study of rubella virus vaccine strain BRD-Ⅱ, so as to fully grasp the sequence characteristics of this genome. MethodsSecond-generation sequencing method was used to conduct the whole-genome sequencing on the vaccine strain BRD-Ⅱ, and the affinity tree of this genome with some vaccine strains and wild-type rubella virus strains was analyzed using the maximum likelihood method. The average genetic distance of nucleic acid sequence of each vaccine strain protein was determined. And homology comparison of structural proteins of each rubella vaccine strain, plus the comparison between this genome with the AY258323.1 genome sequence, were conducted to analyze the homology of E1 protein between the wild-type rubella virus reference strain and vaccine strain BRD-Ⅱ. ResultsThe sequencing results showed that the BRD-Ⅱ strain was a single-molecule single-stranded positive-strand ribonucleic acid (RNA), composed of 9 778 nucleotides, with a GC content of 69.35 %. The C protein was composed of 300 amino acids, the E2 glycoprotein was composed of 282 amino acids, and the E1 glycoprotein was composed of 481 amino acids. The results of preliminary analysis showed that the average genetic distances of nucleic acid sequences were 0.066 700 for the P150 protein, 0.061 933 for the P90 protein, 0.057 850 for the C protein, 0.068 167 for the E2 protein, and 0.068 833 for the E1 protein, respectively. The amino acid sequences in the E2 protein and E1 protein regions of the two BRD-Ⅱ strains did not change, confirming the conserved regions of the E1 protein by comparison. ConclusionThe sequence characteristics of the genome are clarified, which have laid a broad foundation for the subsequent detection of the genetic stability of the main antigen genes.
2.Discussion on Research Paradigm and Layout Based on Disease (Psoriasis) Specifically Responding to Traditional Chinese Medicine
Liu LIU ; Xiaoying SUN ; Mei MO ; Zihan FANG ; Yaqiong ZHOU ; Bin LI ; Xiaoxiao ZHANG ; Xin LI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(18):203-211
To implement the Opinions of the State Council of the Central Committee of the Communist Party of China on Promoting the Inheritance and Innovative Development of Traditional Chinese Medicine, delve into the diseases specifically responding to traditional Chinese medicine (TCM), and serve the building of advantageous specialties, clinical talent cultivation, national research layout, and academic innovation, the Chinese Association of Chinese Medicine sponsored the salons in multiple fields to discuss the diseases specifically responding to TCM. On August 26, 2023, the 25th salon on the diseases specifically responding to TCM was held in Shanghai. In view of the advantages of TCM and integrated traditional Chinese and Western medicine in the diagnosis and treatment of psoriasis, Chinese and western medicine experts and interdisciplinary researchers carried out extensive and in-depth discussions and formed specific suggestions and consensus on the diagnosis and treatment of psoriasis with TCM and integrated traditional Chinese and Western medicine. However, there was still a lack of detailed research path. Under the guidance of the Chinese Association of Chinese Medicine, this paper analyzes the problems in the diagnosis and treatment of psoriasis from the occurrence and development of psoriasis. According to the advantages and characteristics of TCM and integrated traditional Chinese and Western medicine in the diagnosis and treatment of psoriasis, this paper puts forward the main points of research layout for psoriasis from the three aspects of psoriasis diagnosis and treatment, comorbidity prevention, and chronic disease management as follows: ① optimization of the syndrome differentiation system for psoriasis, ② optimization of assessment indicators for psoriasis, ③ recurrence mechanisms of psoriasis, ④ construction and research of TCM prevention and treatment of psoriasis recurrence, ⑤ construction of the comorbidity spectrum and TCM theoretical system of psoriasis, ⑥ comorbidity mechanisms of psoriasis. Furthermore, this article proposed the research layout and directions, expected goals and values, and funding priorities. Therefore, on the basis of the series of salons about psoriasis as a disease specifically responding to TCM, this article puts forward the research paradigm of psoriasis, aiming to facilitate the high-quality development of TCM and provide reference for the national research layout, the research and development of new TCM preparations, the selection of research topics, and the formulation of guidelines and consensus.
3.Yishen Huashi Granules Protect Kidneys of db/db Mice via p38 MAPK Signaling Pathway
Kaidong ZHOU ; Sitong WANG ; Ge JIN ; Yanmo CAI ; Xin ZHOU ; Yunhua LIU ; Xinxue ZHANG ; Min ZHANG ; Zongjiang ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):58-68
ObjectiveTo explore the mechanism of Yishen Huashi granules in alleviating renal tubular epithelial cell injury and relieving diabetic kidney disease by regulating the mitogen-activated protein kinase (MAPK) signaling pathway. MethodsThe db/db mice of 12 weeks old were randomly assigned into model , dapagliflozin (1.6 mg·kg-1), and Yishen Huashi granules (4.7 g·kg-1), and db/m mice were used as the control group. The general conditions of mice were observed, and fasting blood glucose and 24-h urinary protein and albumin-to-creatinine ratio (ACR) were measured at weeks 0 and 12 of administration. After 12 weeks of treatment, the levels of serum creatinine (SCr), blood urea (UREA), triglycerides (TG), total cholesterol (TC), and low density lipoprotein (LDL) were measured. The pathological changes in the renal tissue were observed by hematoxylin-eosin (HE) staining, Periodic acid-Schiff (PAS) staining, Mallory staining, and transmission electron microscopy. Real-time PCR was employed to determine the mRNA levels of monocyte chemotactic protein-1 (MCP-1) and CC chemokine receptor-2 (CCR2) in the renal tissue of mice. The immunohistochemical assay was employed to examine the expression of p38, phospho-p38 (p-p38), MCP-1, and CCR2 in the renal tissue of mice. Western blotting was employed to measure the protein levels of p-p38, p38, MCP-1, and CCR2 in the renal tissue of mice.HK-2 cells cultured in vitro were grouped as follows: negative control, high glucose(30 mmol·L-1), Yishen Huashi granule-containing serum, and SB203580. After 48 h of cell culture in each group, RNA were extracted and the levels of MCP-1, and CCR2 mRNA were determined by Real-time PCR,proteins were extracted and the levels of p38, p-p38, MCP-1, and CCR2 were determined by Western blot. ResultsThe in vivo experiments showed that before treatment, other groups had higher body weight, blood glucose level, 24 h urinary protein, and ACR than the control group (P<0.05,P<0.01). After 12 weeks of treatment, compared with the model group, the Yishen Huashi granules group showed improved general conditions, a decreasing trend in body weight, lowered levels of blood glucose, 24-h urinary protein, and ACR (P<0.01), reduced SCr and UREA (P<0.01), and declined levels of TC, TG, and LDL (P<0.05,P<0.01). Compared with the model group, the Yishen Huashi granules group showed alleviated damage and interstitial fibrosis in the renal tissue as well as reductions in glomerular foot process fusion and basement membrane thickening. Moreover, the Yishen Huashi granules group showed down-regulated mRNA levels of MCP-1 and CCR2 (P<0.01), reduced positive expression of p-p38, MCP-1, and CCR2 (P<0.01), and down-regulated protein levels of p-p38/p38, MCP-1, and CCR2 (P<0.05) in the renal tissue. The cell experiment showed that compared with the high glucose group, the Yishen Huashi granule-containing serum group showcased down-regulated mRNA levels of MCP-1 and CCR2 (P<0.01) and down-regulated protein levels of p-p38/p38, MCP-1, and CCR2(P<0.05,P<0.01). ConclusionYishen Huashi granules can regulate glucose-lipid metabolism, reduce 24 h urinary protein and ACR, improve the renal function, alleviate the renal tubule injury caused by high glucose, and protect renal tubule epithelial cells in db/db mice by reducing MCP-1/CCR2 activation via the p38 MAPK signaling pathway.
4.Clinical Diseases Responding Specially to TCM Treatment: Psoriasis
Liu LIU ; Xiaoying SUN ; Mei MO ; Yaqiong ZHOU ; Bin LI ; Xiaoxiao ZHANG ; Xin LI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(5):260-268
Psoriasis is a common chronic inflammatory systemic disease in dermatology. Its high prevalence, recurrence rate, and numerous comorbidities impose a significant physical and mental burden on patients. With the continuous advancement of modern medicine, the emergence of biological agents has improved clinical efficacy, making it possible to overcome psoriasis, in addition to classical treatments. However, in clinical practice, adverse reactions, drug resistance, recurrence rates, and immune drift cannot be ignored. Traditional Chinese medicine (TCM) has a history of thousands of years in treating psoriasis, demonstrating good efficacy, high safety, and a low recurrence rate, but a standardized management system is lacking. Therefore, the 25th Clinical Diseases Responding Specially to TCM Treatment Series (Psoriasis) Youth Salon, hosted by the Chinese Association of Chinese Medicine and organized by the Youth Committee of the Chinese Association of Chinese Medicine, invited 29 experts and scholars from TCM, Western medicine, and interdisciplinary fields to actively discuss the "Advantages, Challenges, and Clinical Transformation of TCM and Western Medicine in the Diagnosis and Treatment of Psoriasis". The experts at the meeting concluded that the advantages of TCM in the treatment of psoriasis are as follows. Firstly, in the TCM-led treatment plan, TCM's understanding of psoriasis follows the principle of combining the differentiation of disease and syndrome. This approach distinguishes the basic contradiction from the current main contradiction and enables a clear grasp of the dynamic process of psoriasis development. Based on the system of syndrome differentiation and treatment, TCM intervention is applied to address the current main contradiction, and the optimal TCM treatment plan is formulated by combining internal and external treatments. Adhering to the principle of "what is visible outside must be addressed inside", TCM can prevent and treat psoriasis comorbidities early by differentiating syndrome types. Secondly, in the integrated TCM and Western medicine treatment plan, the combination of both methods not only enhances efficacy but also reduces the adverse reactions of immunosuppressants and biological agents, lowering the recurrence rate. This conference provides a reference for the diagnosis and treatment of psoriasis using TCM and integrated TCM and Western medicine, opening up new ideas for clinical and basic research and guiding future research directions.
5.Liver histopathological features of HBeAg-negative patients in the indeterminate phase of low-viral-load chronic hepatitis B virus infection
Lulu ZHOU ; Bing DONG ; Jiejing XIN ; Guanghua XU ; Na LIU
Journal of Clinical Hepatology 2025;41(1):52-56
ObjectiveTo investigate the liver histopathological features of HBeAg-negative patients in the indeterminate phase of low-viral-load chronic hepatitis B virus (HBV) infection. MethodsA total of 271 patients with low-viral-load HBeAg-negative chronic HBV infection who underwent liver biopsy in Department of Infectious Diseases, Affiliated Hospital of Yan’an University, from September 2013 to June 2021 were enrolled as subjects, and the degree of liver injury was compared between patients based on age, sex, presence or absence of the family history of hepatitis B, HBsAg, and alanine aminotransferase (ALT) level. The chi-square test was used for comparison of categorical data between two groups. ResultsAmong the 271 patients with HBeAg-negative chronic HBV infection, 86 patients (31.73%) grade≥A2 liver inflammatory activity, 72 (26.57%) had a liver fibrosis stage of ive, and 112 (41.33%) had moderate or severe liver histological injury. The proportion of patients with grade≥A2 liver inflammatory activity in the patients with ALT>20 U/L was significantly higher than that in the patients with ALT≤20 U/L (χ2=3.938, P=0.047). There were no significant differences in the proportion of patients with grade≥A2 liver inflammatory activity between the patients with different ages, sexes, family history of hepatitis B, HBsAg levels (all P>0.05),there were no significant differences in the proportion of patients with a liver fibrosis stage of ≥F2 between the patients with different ages, sexes, family history of hepatitis B, HBsAg, and ALT levels (all P>0.05), and the stratified analysis of patients aged≤30 years and patients without the family history of hepatitis B showed no statistical significance between groups (all P>0.05). There was no significant difference in the degree of liver histological injury between the patients with different ages, sexes, family history of hepatitis B, HBsAg, and ALT levels (all P>0.05). ConclusionSignificant liver injury is observed in more than 40% of the patients with low-viral-load HBeAg-negative chronic HBV infection, and there is no significant difference in the degree of liver histological injury between the patients with different ages, sexes, family history of hepatitis B, HBsAg, and ALT levels. Even for the patients aged≤30 years who deny the family history of hepatitis B, there is still a considerable proportion of patients with liver injury, which should be taken seriously by clinicians.
6.Structure, content and data standardization of rehabilitation medical records
Yaru YANG ; Zhuoying QIU ; Di CHEN ; Zhongyan WANG ; Meng ZHANG ; Shiyong WU ; Yaoguang ZHANG ; Xiaoxie LIU ; Yanyan YANG ; Bin ZENG ; Mouwang ZHOU ; Yuxiao XIE ; Guangxu XU ; Jiejiao ZHENG ; Mingsheng ZHANG ; Xiangming YE ; Jian YANG ; Na AN ; Yuanjun DONG ; Xiaojia XIN ; Xiangxia REN ; Ye LIU ; Yifan TIAN
Chinese Journal of Rehabilitation Theory and Practice 2025;31(1):21-32
ObjectiveTo elucidate the critical role of rehabilitation medical records (including electronic records) in rehabilitation medicine's clinical practice and management, comprehensively analyzed the structure, core content and data standards of rehabilitation medical records, to develop a standardized medical record data architecture and core dataset suitable for rehabilitation medicine and to explore the application of rehabilitation data in performance evaluation and payment. MethodsBased on the regulatory documents Basic Specifications for Medical Record Writing and Basic Specifications for Electronic Medical Records (Trial) issued by National Health Commission of China, and referencing the World Health Organization (WHO) Family of International Classifications (WHO-FICs) classifications, International Classification of Diseases (ICD-10/ICD-11), International Classification of Functioning, Disability and Health (ICF), and International Classification of Health Interventions (ICHI Beta-3), this study constructed the data architecture, core content and data standards for rehabilitation medical records. Furthermore, it explored the application of rehabilitation record summary sheets (home page) data in rehabilitation medical statistics and payment methods, including Diagnosis-related Groups (DRG), Diagnosis-Intervention Packet (DIP) and Case Mix Index. ResultsThis study proposed a systematic standard framework for rehabilitation medical records, covering key components such as patient demographics, rehabilitation diagnosis, functional assessment, rehabilitation treatment prescriptions, progress evaluations and discharge summaries. The research analyzed the systematic application methods and data standards of ICD-10/ICD-11, ICF and ICHI Beta-3 in the fields of medical record terminology, coding and assessment. Constructing a standardized data structure and data standards for rehabilitation medical records can significantly improve the quality of data reporting based on the medical record summary sheet, thereby enhancing the quality control of rehabilitation services, effectively supporting the optimization of rehabilitation medical insurance payment mechanisms, and contributing to the establishment of rehabilitation medical performance evaluation and payment based on DRG and DIP. ConclusionStructured rehabilitation records and data standardization are crucial tools for quality control in rehabilitation. Systematically applying the three reference classifications of the WHO-FICs, and aligning with national medical record and electronic health record specifications, facilitate the development of a standardized rehabilitation record architecture and core dataset. Standardizing rehabilitation care pathways based on the ICF methodology, and developing ICF- and ICD-11-based rehabilitation assessment tools, auxiliary diagnostic and therapeutic systems, and supporting terminology and coding systems, can effectively enhance the quality of rehabilitation records and enable interoperability and sharing of rehabilitation data with other medical data, ultimately improving the quality and safety of rehabilitation services.
7.Efficacy and Mechanism of Action of Ermiao Situ Decoction in Modulating JAK/STAT Pathway in Rats with Damp-heat Eczema
Kangning HAN ; Junjie HU ; Juan LI ; Min ZHANG ; Xian ZHOU ; Songlin LIU ; Xin CHEN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(9):37-47
ObjectiveUltra performance liquid chromatography-quadrupole-time of flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS) coupled with network pharmacology and molecular docking was utilized to explore the efficacy and mechanism of action of Ermiao Situ decoction on rats with damp-heat eczema. MethodsA rat model of damp-heat eczema was established by artificial climate chamber intervention combined with sensitization induction by dinitrochlorobenzene (DNCB), and it was randomly divided into the normal group, the model group, the medium- and high-dose groups of Ermiao Situ decoction (3.40 g·kg-1 and 6.80 g·kg-1), and the prednisone acetate group (2.51 mg·kg-1), with eight rats in each group, totalling 46 rats, of which six rats were tested with the drug-containing serum. The chemical analysis of drug-containing serum from rats was carried out by UPLC-Q-TOF-MS/MS, combined with network pharmacology for the prediction of key components, core targets, and signaling pathways, and molecular docking experiments were performed by CB-Dock2 online website. The pharmacological effects of Ermiao Situ decoction in the treatment of damp-heat eczema were investigated by epitaxial indexes combined with the pathologic tissue staining method. The serum levels of gastrin (GAS), interleukin-4 (IL-4), and interleukin-13 (IL-13) were measured by enzyme-linked immunosorbent assay (ELISA). Interleukin-6 (IL-6), Janus kinase 1 (JAK1), phosphorylated (p)-JAK1, signal transduction and activation of transcription factor 3 (STAT3), and p-STAT3 protein expression level was determined by Western bolt. ResultsA total of 19 active ingredients were detected in drug-containing serum samples of rats, which were predicted to act on 198 targets for the treatment of damp-heat eczema, among which the key ingredients included rhodopsin, huangpai alkaloids, and quercetin, and the main core targets included STAT3, tumor necrosis factor (TNF), and IL-6, which were mainly involved in the cancer signaling pathway, phosphatidylinositol 3-kinase (PI3K)/protein kinase (Akt) signaling pathway, T helper 17 (Th17) cell differentiation signaling pathway, and JAK/STAT signaling pathway. The molecular docking results suggested that the key components had strong binding activities with the core targets IL-6, JAK1, and STAT3 in the JAK/STAT signaling pathway. The results of animal experiments showed that compared with those in the normal group, rats in the model group were depressed. They had loose hair, loose stools, epidermal oozing, vesiculation, and generation of thick scabs in the form of scales, decreased body weight, increased anus temperature and water intake, and increased indexes of the spleen, thymus gland, and stomach (P<0.05, P<0.01), and the lesion tissue could be seen to be hyperkeratotic, with the aggregation of inflammatory cells and nonsignificant separation of epidermis and dermis. The gastric mucosa was thinned, deficient, and structurally disorganized, and obvious inflammatory cell aggregation was seen. The levels of GAS, IL-4, and IL-13 in serum were significantly reduced (P<0.05, P<0.01), and the protein expression levels of IL-6, JAK1, p-JAK1, and p-STAT3 in the lesion tissue were significantly increased (P<0.05, P<0.01). Compared with those in the model group, rats in each administration group had stable mental states, formed feces, a clean perianal area, and basically normal epidermis. Only a small amount of scaly scabs existed, and the rats had body weight increased, with decreased anal temperature and water intake, as well as decreased spleen, thymus, and gastric indexes (P<0.05, P<0.01). Epidermal thickness was decreased, and epidermal and dermal separation boundaries were obvious, but hyperkeratotic and accumulation of inflammatory cells could still be seen. The thickness of gastric mucosa increased, and the structure was restored to varying degrees. The levels of GAS, IL-4, and IL-13 content in the serum of rats were increased to varying degrees, and the protein expression levels of IL-6, JAK1, p-JAK1, and p-STAT3 in the dermal lesion tissue were significantly decreased (P<0.05, P<0.01). ConclusionErmiao Situ decoction may exert therapeutic effects on rats with damp-heat eczema by modulating the JAK/STAT signaling pathway.
8.Process Optimization and Health Risk Assessment of Calcined Haematitum Based on QbD Concept
Yue YANG ; Jingwei ZHOU ; Jialiang ZOU ; Guorong MEI ; Yifan SHI ; Lei ZHONG ; Jiaojiao WANG ; Xuelian GAN ; Dewen ZENG ; Xin CHEN ; Lin CHEN ; Hongping CHEN ; Shilin CHEN ; Yuan HU ; Youping LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(13):187-196
ObjectiveTo investigate the processing technology of calcined Haematitum based on the concept of quality by design(QbD) and to assess its health risk. MethodsTaking whole iron content, Fe2+ dissolution content and looseness as critical quality attributes(CQAs), and calcination temperature, calcination time, spreading thickness and particle size as critical process parameters(CPPs) determined by the failure mode and effect analysis(FMEA), the processing technology of calcined Haematitum was optimized by orthogonal test combined with analytic hierarchy process-criteria importance through intercriteria correlation(AHP-CRITIC) hybrid weighting method. The contents of heavy metals and harmful elements were determined by inductively coupled plasma mass spectrometry, and the health risk assessment was carried out by daily exposure(EXP), target hazard quotient(THQ) and lifetime cancer risk(LCR), and the theoretical value of the maximum limit was deduced. ResultsThe optimal processing technology for calcined Haematitum was calcination at 650 ℃, calcination time of 1 h, particle size of 0.2-0.5 cm, spreading thickness of 1 cm, and vinegar quenching for 1 time[Haematitum-vinegar(10:3)]. The contents of 5 heavy metals and harmful elements in 13 batches of calcined Haematitum were all decreased with reductions of up to 5-fold. The cumulative THQ of 2 batches of samples was>1, while the cumulative THQ of all batches of Haematitum was>1. The LCR of As in 1 batches of Haematitum was 1×10-6-1×10-4, and the LCR of the rest was<1×10-6, and the LCRs of calcined Haematitum were all<1×10-6, indicating that the carcinogenic risk of calcined Haematitum was low, but special attention should still be paid to Haematitum medicinal materials. Preliminary theoretical values of the maximum limits of Cu, As, Cd, Pb and Hg were formulated as 1 014, 25, 17, 27, 7 mg·kg-1. ConclusionThe optimized processing technology of calcined Haematitum is stable and feasible, and the contents of heavy metals and harmful elements are reduced after processing. Preliminary theoretical values of the maximum limits of Cu, As, Cd, Pb and Hg are formulated to provide a scientific basis for the formulation of standards for the limits of harmful elements in Haematitum.
9.Optimization of Processing Technology of Calcined Pyritum Based on QbD Concept and Its XRD Fingerprint Analysis
Xin CHEN ; Jingwei ZHOU ; Haiying GOU ; Lei ZHONG ; Tianxing HE ; Wenbo FEI ; Jialiang ZOU ; Yue YANG ; Dewen ZENG ; Lin CHEN ; Hongping CHEN ; Shilin CHEN ; Yuan HU ; Youping LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(13):197-205
ObjectiveBased on the concept of quality by design(QbD), the processing process of calcined Pyritum was optimized, and its X-ray diffraction(XRD) fingerprint was established. MethodsThe safety, effectiveness and quality controllability of calcined Pyritum were taken as the quality profile(QTPP), the color, hardness, metallic luster, phase composition, the contents of heavy metals and hazardous elements were taken as the critical quality attributes(CQAs), and the calcination temperature, calcination time, paving thickness and particle size were determined as the critical process parameters(CPPs). Differential thermal analysis, X-ray diffraction(XRD) and inductively coupled plasma mass spectrometry(ICP-MS) were used to analyze the correlation between the calcination temperature and CQAs of calcined Pyritum. Then, based on the criteria importance through intercriteria correlation(CRITIC)-entropy weight method, the optimal processing process of calcined Pyritum was optimized by orthogonal test. Powder XRD was used to analyze the phase of calcined Pyritum samples processed according to the best process, and the mean and median maps of calcined Pyritum were established by the superposition of geometric topological figures, and similarity evaluation and cluster analysis were carried out. ResultsThe results of single factor experiments showed that the physical phase of Pyritum changed from FeS2 to Fe7S8 during the process of temperature increase, the color gradually deepened from dark yellow, and the contents of heavy metals and harmful elements decreased. The optimized processing process of calcined Pyritum was as follows:calcination temperature at 750 ℃, calcination time of 2.5 h, paving thickness of 3 cm, particle size of 0.8-1.2 cm, vinegar quenching 1 time[Pyritum-vinegar(10∶3)]. After calcination, the internal structure of Pyritum was honeycomb-shaped, which was conducive to the dissolution of active ingredients. XRD fingerprints of 13 batches of calcined Pyritum characterized by 10 common peaks were established. The similarities of the relative peak intensities of the XRD fingerprints of the analyzed samples were>0.96, and it could effectively distinguish the raw products and unqualified products. ConclusionTemperature is the main factor affecting the quality of calcined Pyritum. After processing, the dissolution of the effective components in Pyritum increases, and the contents of heavy metals and harmful substances decrease, reflecting the function of processing to increase efficiency and reduce toxicity. The optimized processing process is stable and feasible, and the established XRD fingerprint can be used as one of the quality control standards of calcined Pyritum.
10.Effect of Shenkang Injection on Podocyte Apoptosis and GRP78/CHOP Signaling Pathway in db/db Mice with Diabetic Kidney Disease Based on Endoplasmic Reticulum Stress
Yanmo CAI ; Sitong WANG ; Xin ZHOU ; Ge JIN ; Kaidong ZHOU ; Yunhua LIU ; Fengfeng ZHANG ; Xinxue ZHANG ; Zongjiang ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):81-90
ObjectiveTo investigate the mechanism of Shenkang injection in delaying diabetic kidney disease by regulating endoplasmic reticulum stress and attenuating podocyte apoptosis through the Glucose regulated protein 78 ( GRP78 ) / transcription factor C / EBP homologous protein ( CHOP ) signaling pathway (GRP78/CHOP) signaling pathway. MethodsFor the animal experiment, 10 12-week-old db/m mice were selected as a normal group, and 30 12-week-old db/db mice were randomly divided into a model group, a Shenkang injection group (15.6 mL·kg-1), and a dapagliflozin group (1.6 mg·kg-1). To observe the general condition of mice, fasting blood glucose, urinary albumin/urine creatinine (ACR), and 24 h urine protein quantification were detected in each group before drug administration. After 12 weeks of drug treatment, mice were tested for fasting blood glucose, total cholesterol (TC), triglyceride (TG), low-density cholesterol (LDL), ACR, 24 h urine protein quantification, blood creatinine (SCr), and blood urea (UREA). Hematoxylin-eosin (HE) staining, periodic acid-Schiff (PAS) staining, and transmission electron microscopy were used to observe the pathologic morphology in renal tissue. Immunohistochemistry was used to detect the expressions of nephroprotective marker protein (Nephrin), glucose-regulated protein 78 (GRP78), CCAAT/enhancer-binding protein homologous protein (CHOP), B-cell lymphoma-2 (Bcl-2), and Bcl-2-associated X protein (Bax) in renal tissue. Western blot was used to detect the expressions of GRP78, CHOP, Bcl-2, Bax, and Nephrin proteins, and Real-time polymerase chain reaction (Real-time PCR) was employed to detect the expressions of Nephrin, GRP78, CHOP, Bcl-2, and Bax mRNAs in renal tissue. ResultsBefore drug administration, compared with those in the normal group, the body mass of db/db mice was significantly increased, and blood glucose, 24 h urine protein quantification, and ACR were significantly elevated in the Shenkang injection group and Dapagliflozin group (P<0.01). After 12 weeks of administration, compared with those in the model group, the general state of mice in the Shenkang injection group was significantly improved, and the body mass was decreased. The blood glucose was significantly reduced (P<0.01), and blood lipids TC, TG, and LDL were significantly decreased (P<0.05, P<0.01). The 24 h urine protein quantification and ACR were significantly decreased (P<0.05), and SCr and UREA were significantly reduced (P<0.01). Compared with those of the model group, the pathologic results of the Shenkang injection group showed that proliferation of mesangial cells, reduction of inflammatory cell infiltration, and alleviation of renal tubular vacuolization and podocyte damage were observed in renal tissue of mice. Electron microscopy showed that fusion of the pedicle protruding and thickening of the basement membrane were reduced. Immunohistochemistry results showed that the expressions of GRP78, CHOP, and Bax proteins were significantly reduced (P<0.01), and the expressions of Nephrin and Bcl-2 proteins were significantly increased (P<0.01) in renal tissue of the Shenkang injection group. Western blot results showed that the expressions of Nephrin and Bcl-2 in the Shenkang injection group were significantly increased (P<0.05, P<0.01), and the expressions of GRP78, CHOP, and Bax proteins were significantly decreased (P<0.05, P<0.01). Real-time PCR results showed that the expressions of GRP78, CHOP, and Bax mRNAs were down regulated in the Shenkang injection group (P<0.01), and the expressions of Nephrin and Bcl-2 mRNAs were up regulated (P<0.01). ConclusionShenkang injection inhibits endoplasmic reticulum stress response and podocyte apoptosis by regulating the GRP78/CHOP signaling pathway, which in turn ensures the integrity of glomerular filtration barrier, reduces the occurrence of proteinuria, improves renal function, and thus delays the progression of diabetic kidney disease.

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