Yigongsan， derived from QIAN Yi’s Key to Therapeutics of Children’s Diseases in the Song Dynasty， is a classic pediatric prescription that is included in the Catalogue of Ancient Classic Prescriptions （the Second Batch of Pediatrics） released by the National Administration of Traditional Chinese Medicine in 2022. This paper verifies and analyzes the historical origin， composition， dosage， processing， decoction method and efficacy of Yigongsan by systematically combing ancient books and modern documents. As a result， Yigongsan is composed of five herbs： Panax ginseng， Poria cocos， Atractylodes macrocephala， Citrus reticulata and Glycyrrhiza uralensis， of which P. cocos should be peeled， A. macrocephala is fried with soil， G. uralensis is roasted with honey while P. ginseng and C. reticulata are raw products. According to the dosage of ancient and modern times， each medicinal herb must be ground into fine powder， 1.6 g for each， added with 300 mL of water， 5 pieces of Zingiber officinale， and 2 Ziziphus jujuba， decocted together to 210 mL， and taken before meals. In ancient books， Yigongsan is used to treat vomiting， diarrhea， spleen and stomach deficiency， chest and abdominal distension， and lack of appetite， etc. Modern research showed that Yigongsan could also be used in the diseases of immune system， respiratory system， blood system， etc.， involving infantile anorexia， asthma， anemia， tumors and so on.

Objective To analyze the quality of 19 batches of Ginseng Formula Granules from 11 different manufacturers by using Ginseng water Extract reference substance(GWERS)as references.Methods Thin layer chromatography(TLC)identification and feature map detection were carried out according to the identification items and characteristic maps of Ginseng Formula Granules standard(YBZ-PFKL-2021186)issued by the National Medical Products Administration.Results The results of TLC analysis showed that the 19 batches of Ginseng Formula Granules-labeled samples could be divided into three categories.The overall pattern of the first type of samples was consistent with that of GWERS,and the similarity was high.Pseudoginsenoside F11,a unique component of American ginseng,was detected in the second type of samples.Four blue fluorescent spots were observed in the third type of samples compared to GWERS.HPLC results indicated that all 19 batches of Ginseng Formula Granules showed eight characteristic peaks at the same retention time as that of GWERS chromatogram.Two more chromatographic peaks were found in the chromatogram of three batches of samples from one manufacturer compared to the chromatogram of other samples,whose similarity to the GWERS chromatogram was less than 0.65.The similarity between the chromatogram of the remaining 16 sample and GWERS chromatogram was higher than 0.94.Conclusion At present,the quality of Ginseng Formula Granules on the market varies greatly.It was suspected that American ginseng might appear among them.The application of GWERS for quality analysis of Ginseng Formula Granules has better applicability to the control medicinal materials.

Objective To investigate the prognostic value of N6-methyladenosine (m⁶A) modification related genes and immune infiltration in colorectal cancer (CRC) and construct a risk model for predicting outcome of patients. Methods The transcriptome data and matched clinical information of CRC patients were downloaded from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) database. The prognostic value of m⁶A modification related genes and immune infiltration were investigated using the consensus clustering method and single sample gene set enrichment analysis. The weighted gene co-expression network analysis (WGCNA) was used to identify prognostic genes related with m⁶A modification and immune infiltration. Lasso regression analysis was used to construct a multi-gene risk model. The expression differences of prognostic genes identified were further validated through expression differential analysis in the Gene Expression Profiling Interactive Analysis (GEPIA) database. Finally, the Kaplan-Meier was used to evaluate the predicting performance of the model in different subgroups and external validation cohorts. Results Both the m⁶A modification and immune infiltration phenotype could effectively stratify the prognosis of CRC patients from the TCGA cohort. Most m⁶A modification related genes were significantly correlated with immune infiltration in CRC tissues. Four following prognostic genes were selected using the WGCNA method combined with Lasso regression analysis: intelectin-1, lymphocyte antigen 6 complex locus G6D, atonal homolog 1 and matrix metalloproteinase 28. In colorectal cancer tissues, the expression levels of lymphocyte antigen 6 complex locus G6D and matrix metalloproteinase 28 exhibited significant differences compared to adjacent non-cancerous tissues (P < 0.05). The risk model constructed based on the above prognostic genes can effectively identify the potential risk population with poor prognosis in different clinical subgroups of the TCGA cohort and the GEO validated cohort. Conclusion A risk model based on m⁶A modification and immune infiltration could effectively predict the clinical outcome of CRC patients, and related prognostic genes have potential to be developed as molecular targets for CRC therapy.

The classic formula Zhulingtang, derived from the medical work Treatise on Cold Damage (《伤寒论》) compiled by ZHANG Zhongjing, a medical sage in the Eastern Han Dynasty, has been included in the Catalogue of Ancient Classic Formulas (First Batch) published by the National Administration of Traditional Chinese Medicine in 2018. Using the method of textual research, this study systematically reviewed ancient and modern literature to conduct a historical and textual analysis of Zhulingtang, including its origin, composition, dosage, processing, decoction methods, efficacy, and applications. A total of 733 pieces of relevant information related to Zhulingtang were collected, involving 206 ancient Chinese medical texts, with 52 of them providing detailed records of the composition, dosage, processing, and efficacy of Zhulingtang. The results of the analysis showed that Zhulingtang was composed of Polyporus, Poria, Alismatis Rhizoma, Asini Corii Colla, and Talcum. Polyporus and Poria should be used without their peels, Asini Corii Colla should be stir-fried with clam powder, and Talcum should be ground into powder or soaked in water. Based on the conversion of ancient and modern dosages, Polyporus, Poria, Alismatis Rhizoma, and Talcum, 15.63 g for each one, were decocted with 800 mL of water to 400 mL. Then the drug residue was removed, and 15.63 g of Asini Corii Colla was added to the drug juice for melting by heating. The decoction should be taken warm, 140 mL each time, three times a day. Zhulingtang has the effects of promoting diuresis, nourishing yin, and clearing heat, and it is mainly used to treat water-heat combination syndrome, characterized by symptoms such as difficult urination, fever, and thirst. Modern research indicates that Zhulingtang is commonly used to treat diseases such as cirrhotic ascites, chronic glomerulonephritis, nephrotic syndrome, diarrhea, and urinary tract infections. This study provides key information about the famous formula Zhulingtang, which can serve as a reference for further development and research on its application.

Evaluating the consistency of herb injectable formulations could improve their product quality and clinical safety, particularly concerning the composition and content levels of trace ingredients. Panax notoginseng Saponins Injection (PNSI), widely used in China for treating acute cardiovascular diseases, contains low-abundance (10%-25%) and trace saponins in addition to its five main constituents (notoginsenoside R₁, ginsenoside Rg₁, ginsenoside Re, ginsenoside Rb₁, and ginsenoside Rd). This study aimed to establish a robust analytical method and assess the variability in trace saponin levels within PNSI from different vendors and formulation types. To achieve this, a liquid chromatography-triple quadrupole mass spectrometry (LC-MS/MS) method employing multiple ions monitoring (MIM) was developed. A "post-column valve switching" strategy was implemented to eliminate highly abundant peaks (NR₁, Rg₁, and Re) at 26 min. A total of 51 saponins in PNSI were quantified or relatively quantified using 18 saponin standards, with digoxin as the internal standard. This study evaluated 119 batches of PNSI from seven vendors, revealing significant variability in trace saponin levels among different vendors and formulation types. These findings highlight the importance of consistent content in low-abundance and trace saponins to ensure product control and clinical safety. Standardization of these ingredients is crucial for maintaining the quality and effectiveness of PNSI in treating acute cardiovascular diseases.
Ginsenosides ; Saponins ; Chemometrics ; Panax notoginseng ; Cardiovascular Diseases ; Chromatography, Liquid ; Tandem Mass Spectrometry

Objective:To analyze the viral genome sequence of novel coronavirus infected persons in Baotou City, understand the mutation characteristics of novel coronavirus genome in the process of transmission among cases, and explore the transmission rule of novel coronavirus in the clustered populations.Methods:Nine throat swabs samples (No. 1 - 7, No. 9, and No. 10), two sputum samples (No. 8, No. 11, and No. 11 sample was from No. 10 case), and one surface smear sample (No.12, and No. 12 sample was from No. 10 case) were collected from 10 confirmed cases of novel coronavirus infection in Baotou City from January 25 to February 21, 2020. Samples 1 and 3 were from single cases, and the rest were from clustered cases. The virus genome was sequenced by metagenomic next-generation sequencing (mNGS), and single nucleotide polymorphism (SNP) mutation sites were screened by comparing with NC_045512, a reference strain of novel coronavirus. Combined with relevant epidemiological information, gene mutation, virus typing, and evolutionary traceability analysis were carried out.Results:The results of viral genome mNGS showed that 76 SNP mutation sites were detected in 12 samples compared with the reference strain NC_045512, including 3 (3.95%) transitions and 73 (96.05%) reversals. There were 19 (25.00%) synonymous mutations and 57 (75.00%) non-synonymous mutations. The analysis of nucleotide and amino acid variation sites showed that mutations were found at five sites (T2821C, C6548T, T16464C, G16858A and T251C) in all the clustered cases (cases 2, 4 - 10). In the single cases, sample 1 had mutations at C9245T and A15340T, and sample 3 had mutation at C13T. The virus typing analysis showed that the samples 1 and 3 belonged to the L type of novel coronavirus, while the rest belonged to the S type of novel coronavirus. The results of genomic evolutionary relationship analysis showed that all the samples could be divided into two branches. The branches of sample 1 and 3 belonged to single cases, and the rest belonged to family clustered cases.Conclusion:The genomic characteristics of the clustered cases of novel coronavirus infection in Baotou City are basically consistent with the epidemiological investigation results, and the transmission of the virus is mainly related to close contact and family gathering.

Pheretima,also called"earthworms",is a well-known animal-derived traditional Chinese medicine that is extensively used in over 50 Chinese patent medicines(CPMs)in Chinese Pharmacopoeia(2020 edi-tion).However,its zoological origin is unclear,both in the herbal market and CPMs.In this study,a strategy for integrating in-house annotated protein databases constructed from close evolutionary relationship-sourced RNA sequencing data from public archival resources and various sequencing al-gorithms(restricted search,open search,and de novo)was developed to characterize the phenotype of natural peptides of three major commercial species of Pheretima,including Pheretima aspergillum(PA),Pheretima vulgaris(PV),and Metaphire magna(MM).We identified 10,477 natural peptides in the PA,7,451 in PV,and 5,896 in MM samples.Five specific signature peptides were screened and then validated using synthetic peptides;these demonstrated robust specificity for the authentication of PA,PV,and MM.Finally,all marker peptides were successfully applied to identify the zoological origins of Brain Heart capsules and Xiaohuoluo pills,revealing the inconsistent Pheretima species used in these CPMs.In conclusion,our integrated strategy could be used for the in-depth characterization of natural peptides of other animal-derived traditional Chinese medicines,especially non-model species with poorly annotated protein databases.

Objective:To establish a questionnaire to assess symptom of embolization syndrome after transcatheter hepatic arterial chemoembolization (TACE) in patients with liver cancer, so as to provide a tool of assessing and managing symptom management after TACE.Methods:From March 2020 to June 2021, through literature review, qualitative interview and Delphi expert consultation, the first draft of symptom assessment questionnaire for TACE post-operative embolism syndrome was prepared. The reliability and validity of the questionnaire were tested in 200 patients with liver cancer treated by TACE in department of Liver Oncology, Zhongshan Hospital affiliated to Fudan University.Results:According to the feedback from Delphi expert consultation a draft questionnaire with 9 items of physiological symptoms and 6 items of psychological and social symptoms was formed. Item analysis showed that each item in the questionnaire had a good degree of differentiation. There was significant correlation between each item and the total score of the questionnaire. Three factors were extracted by exploratory factor analysis, naming psychosocial symptom group as factor 1, somatic discomfort symptom group as factor 2 and gastrointestinal reaction symptom group as factor 3, and the cumulative variance contribution rate was 62.592%. Spearman correlation coefficient between this questionnaire and the Anderson Symptom Assessment Scale was 0.855( P<0.05). The Cronbach α of the total questionnaire was 0.898, and. The Cronbach α of the three factors were 0.885, 0.771 and 0.870 respectively. Conclusions:The symptom assessment questionnaire of embolization syndrome after TACE in liver cancer patients prepared in this study has good reliability and validity, which can provide an evaluation basis for the symptom management of TACE.

Objective:To evaluate the detection ability of two kinds of high-throughput method to determine neutralizing antibodies based on a microneutralization test (MNT).Methods:Serum samples were collected from the early phase and follow-up period (117 samples in total) for neutralizing antibody testing. They were tested using MNT, pseudovirus neutralization assay (PBNA), competitive inhibition assay (including enzyme-linked immunosorbent assay (ELISA) and chemiluminescence immunoassay (CLIA)) to evaluate the correlation coefficients and threshold values for the effectiveness of these high-throughput neutralizing antibody assays.Results:The correlation coefficients for PBNA, ELISA, and CLIA relative to MNT were 0.760, 0.778, and 0.725, respectively, for individuals infected with 2019-nCoV. The area under the ROC curve was 0.901 for a cutoff value of 50 for the PBNA assay, 0.934 for a cutoff value of 1∶8 for the ELISA assay and 0.838 for a cutoff value of 1.28AU/ml for the CLIA assay when the threshold value for the microneutralization test was taken as 1: 10 (less than 1: 10 is considered negative).Conclusions:The high-throughput method for the detection of COVID-19 neutralizing antibodies are scientific and feasible, and provide an important technical tool for the regular prevention and control of the epidemic.

Objective:To investigate the expressions of human epidermal growth factor receptor 2 (HER2) in recurrent patients after radical gastrectomy and its correlation with clinicopathologic features and prognosis.Methods:The clinical data of 116 recurrent patients after radical gastrectomy between December 2011 and March 2019 in Fujian Cancer Hospital were retrospectively analyzed. The relationship between HER2 expression of the tissues after radical gastrectomy and clinicopathological features as well as prognosis was also analyzed.Results:HER2 positive rate was 19.8% (23/116) in gastric cancer specimens of 116 patients after radical gastrectomy. HER2 positive rate in highly and moderately differentiated patients with gastric cancer was higher than that in those with poorly differentiated gastric cancer [33.3% (11/33) vs. 14.5% (12/83), χ2 = 5.292, P < 0.05]. HER2 positive rate in patients with lung metastasis was higher than that in patients without lung metastasis [42.1% (8/19) vs. 15.5% (15/97), χ2 = 5.517, P < 0.05]. There were no statistical differences in HER2 positive rate among gastric cancer patients with different gender, age, tumor location, recurrent site, TNM stage, number of metastasis, neurovascular invasion, liver metastasis, local recurrence (all P > 0.05). HER2 expression was not associated with disease-free survival time ( P > 0.05). Conclusion:HER2 expression is associated with the differentiation degree of gastric cancer and the location of distant metastasis, but it can not be used as a predictor for recurrence of gastric cancer.