Benzylisoquinoline alkaloids (BIAs) are a structurally diverse group of plant metabolites renowned for their pharmacological properties. However, sustainable sources for these compounds remain limited. Consequently, researchers are focusing on elucidating BIA biosynthetic pathways and genes to explore alternative sources using synthetic biology approaches. CYP80B, a family of cytochrome P450 (CYP450) enzymes, plays a crucial role in BIA biosynthesis. Previously reported CYP80Bs are known to catalyze the 3'-hydroxylation of (S)-N-methylcoclaurine, with the N-methyl group essential for catalytic activity. In this study, we successfully cloned a full-length CYP80B gene (StCYP80B) from Stephania tetrandra (S. tetrandra) and identified its function using a yeast heterologous expression system. Both in vivo yeast feeding and in vitro enzyme analysis demonstrated that StCYP80B could catalyze N-methylcoclaurine and coclaurine into their respective 3'-hydroxylated products. Notably, StCYP80B exhibited an expanded substrate selectivity compared to previously reported wild-type CYP80Bs, as it did not require an N-methyl group for hydroxylase activity. Furthermore, StCYP80B displayed a clear preference for the (S)-configuration. Co-expression of StCYP80B with the CYP450 reductases (CPRs, StCPR1, and StCPR2), also cloned from S. tetrandra, significantly enhanced the catalytic activity towards (S)-coclaurine. Site-directed mutagenesis of StCYP80B revealed that the residue H205 is crucial for coclaurine catalysis. Additionally, StCYP80B exhibited tissue-specific expression in plants. This study provides new genetic resources for the biosynthesis of BIAs and further elucidates their synthetic pathway in natural plant systems.
Cytochrome P-450 Enzyme System/chemistry* ; Benzylisoquinolines/chemistry* ; Hydroxylation ; Plant Proteins/chemistry* ; Alkaloids/metabolism* ; Stephania tetrandra/genetics*

In the treatment of diabetic gastroparesis(DGP),traditional Chinese medicine has conventionally focused on therapies such as ascending clear and descending turbid,pungent-opening and bitter-descending methods,aiming to regulate the ascending and descending of the spleen and stomach's Qi mechanism and to restore the middle-jiao's transformation as the ultimate goal.By explo-ring the physiological relationship between gallbladder and spleen-stomach and its pathological relationship with DGP,this article sug-gests that the gallbladder also participates in the regulation of blood sugar and digestive activities in the body,which is closely related to the onset of the disease.As the Shao Yang pivot,the gallbladder stores essential fluid,harbors ministerial fire,primarily governs the upward movement and dispersion,and is responsible for decision-making and emotions.The normal flow of Qi,blood,and body flu-ids,as well as the functional activities of the channels and collateral vessels,are closely associated with the gallbladder.When the gallbladder's function becomes abnormal,the body may also exhibit imbalances in the intestinal microbiota,disarray of gastrointestinal hormones,delayed gastric emptying,and elevated blood sugar levels,which aligns with the modern medical understanding of the onset of gastroparesis diabeticorum.Therefore,this article proposes the treatment principle of"venting the wood constraint"and the treatment of DGP according to symptoms,offering a reference for clinical treatment.

Objective:Telemedicine,as an information-based tool,is widely recognized as an effective solution for compensating for the imbalanced allocation of medical resources in China.This study specifi-cally aimed to analyze the impact of telemedicine functions on the operational efficiency of public hospi-tals,with a particular focus on their heterogeneous effects on hospitals of different levels.Methods:A cross-sectional research design was used based on the 2022 Health Informatization Statistical Survey data,and 8 944 public hospitals were used as research objects to analyze the impact of telemedicine on hospital revenues and business capacity.Multivariate linear model,propensity score matching(PSM),and grouped regression methods were employed to evaluate the impact of telemedicine on hospital revenues,number of consultations,and the number of discharges.Results:The descriptive results showed that telemedicine was available in 35.51％of public hospitals.The analysis also demonstrated that various factors,such as hospital level,academic category,area of the hospital,administrational level and num-ber of beds all had a significant influence on the operation of the hospital.Moreover,the regression re-sults showed that opening telemedicine could increase hospital revenues by 0.140(P＜0.01),hospital consultations by 0.136(P＜0.01),and the number of discharges by 0.316(P＜0.01).After correc-ting for endogeneity using the propensity score matching,the results showed that the effect of opening telemedicine on hospital revenues,consultations,and the number of discharges was 0.191(P＜0.01),0.216(P＜0.01),and 0.353(P＜0.01),respectively.Further heterogeneity analysis was conducted to explore the differential effects of telemedicine on hospitals of different levels.Grouped regression showed that telemedicine had a positive impact on the income of secondary hospitals,with a coefficient of 0.088(P＜0.05),and it had a more significant positive impact on hospital consultations in secondary hospitals,with a coefficient of 0.127(P＜0.01).An even greater impact on the number of discharges in primary hospitals,with a coefficient of 1.203(P＜0.01).Telemedicine,on the other hand,did not have a significant positive impact on the overall revenue and operational capacity of tertiary hospitals.Conclusion:Telemedicine had a significant promoting effect on hospital revenues,hospital consultations and the number of discharges,and this effect was differentiated between hospitals of different levels.Through the construction of telemedicine,primary hospitals were able to significantly improve their business capacity and revenue,which played a positive role in improving the operation of primary public hospitals.

Objective:To analyze the clinical and imaging features of patients with COVID-19-related osmotic demyelination syndrome (ODS).Methods:COVID-19-related ODS cases diagnosed in the Department of Neurology, Peking Union Medical College Hospital from January 2020 to September 2023 were retrospectively reviewed. And their past medical history, possible triggers, clinical manifestations, imaging manifestations, treatment and prognosis were summarized.Results:A total of 5 patients with COVID-19-related ODS were included. Electrolyte disturbances acted as an inducement of ODS in all patients (5/5),4 of whom with hyponatremia. Four of 5 patients first presented with disturbance of consciousness, followed by predominant dystonia. Imaging of all patients (5/5) showed isolated extrapontine myelinolysis (EPM). With the prolongation of the course of disease, such signal intensity could return to normal, and lesions showed atrophic changes in some patients. The patients′ clinical symptoms were partly relieved within a few days to a few months after treatment.Conclusions:COVID-19-related ODS is mostly associated with hyponatremia, and EPM is more common. COVID-19 should be considered as a risk factor for ODS.

Purpose: Radiation-induced dermatitis (RD) is a common side-effect of therapeutic ionizing radiation that can severely affect patient quality of life. This study aimed to develop a risk prediction model for the occurrence of RD in patients with cervical carcinoma undergoing chemoradiotherapy using electronic medical records (EMRs). Methods: Using EMRs, the clinical data of patients who underwent simultaneous radiotherapy and chemotherapy at a tertiary cancer hospital between 2017 and 2022 were retrospectively collected, and the patients were divided into two groups: a training group and a validation group. A predictive model was constructed to predict the development of RD in patients who underwent concurrent radiotherapy and chemotherapy for cervical cancer. Finally, the model's efficacy was validated using a receiver operating characteristic curve. Results: The incidence of radiation dermatitis was 89.5% (560/626) in the entire cohort, 88.6% (388/438) in the training group, and 91.5% (172/188) in the experimental group. The nomogram was established based on the following factors: age, the days between the beginning and conclusion of radiotherapy, the serum albumin after chemoradiotherapy, the use of single or multiple drugs for concurrent chemotherapy, and the total dose of afterloading radiotherapy. Internal and external verification indicated that the model had good discriminatory ability. Overall, the model achieved an area under the receiver operating characteristic curve of .66. Conclusions The risk of RD in patients with cervical carcinoma undergoing chemoradiotherapy is high. A risk prediction model can be developed for RD in cervical carcinoma patients undergoing chemoradiotherapy, based on over 5 years of EMR data from a tertiary cancer hospital.

This article summarizes the clinical experience in treatment of refractory tinnitus with He's santong therapy of acupuncture (three techniques of acupuncture to remove the blockage and promote circulation). Refractory tinnitus is characterized by a mixture of deficiency and excess, which is implicated in the dysfunction of zangfu organs, resulting in physical and mental injuries. In association with the complicated conditions of the diseases and etiology, He's santong therapy of acupuncture is adopted. The needling with filiform needles acts directly on the affected site to regulate the mind and improve the hearing, the fire needling works on removing the refractory illness and blockage through heating up, and bloodletting on activating blood circulation and promoting qi flowing to eliminate the pathogens. The three techniques of acupuncture co-work on harmonizing the mind, promoting the circulations in the ear to stop tinnitus.
Humans ; Tinnitus/physiopathology* ; Acupuncture Therapy ; Male ; Middle Aged ; Adult ; Female ; Acupuncture Points

OBJECTIVE Designing and synthesizing ursolic acid prodrugs to improve the water solubility and anti-multiple sclerosis(MS) activity of ursolic acid. METHODS succinic acid, dipeptide amino acid and the amino acids segments of side chains of isavuconazole wers selected according to the prodrug strategy, combined with 3-position hydroxyl of ursolic acid via condensation and esterification reaction. Using ultraviolet-visible spectrophotometry to determine the water solubility of prodrugs, and establish the experimental allergic encephalomyelitis mice model for its anti-MS efficacy evaluation. RESULTS Three prodrugs were synthesized and their structures were confirmed by 1H-NMR, 13C-NMR and LCMS(ESI). The water solubility of prodrugs 1 and 2 was over 110 times higher than ursolic acid, and that of prodrug 3 was over 200 times higher than ursolic acid. The in vivo anti-MS activity results showed that the three prodrugs showed significant anti-MS activity, among which prodrug 1 showed significantly better anti-MS activity than ursolic acid group. The mechanism might be through inhibition of peripheral inflammatory cell infiltration into the center and anti-demyelination, so as to exert the anti-MS activity. The metabolism of prodrug 1 in vivo showed that the exposure of prodrug 1 in vivo was 6 times that of ursolic acid, indicating that prodrug 1 might exert anti-MS activity mainly as sodium prototype. CONCLUSION Prodrug 1 has great anti-MS potential, which is worthy of further study. This study provides some basis and useful guidance for the development of ursolic acid prodrugs with anti-MS activity.

This study aimed to investigate the effect and mechanisms of Ephedra Herb (EH) extract on adriamycin-induced nephrotic syndrome (NS), providing an experimental basis for the clinical treatment of NS. Hematoxylin and eosin staining, creatinine, urea nitrogen, and kidn injury molecule-1 were used to evaluate the activities of EH extract on renal function. The levels of inflammatory factors and oxidative stress were detected by kits. The levels of reactive oxygen species, immune cells, and apoptosis were measured by flow cytometry. A network pharmacological approach was used to predict the potential targets and mechanisms of EH extract in the treatment of NS. The protein levels of apoptosis-related proteins and CAMKK2, p-CAMKK2, AMPK, p-AMPK, mTOR and p-mTOR in the kidneys were detected by Western blot. The effective material basis of EH extract was screened by MTT assay. The AMPK pathway inhibitor (compound C, CC) was added to investigate the effect of the potent material basis on adriamycin-induced cell injury. EH extract significantly improved renal injury and relieve inflammation, oxidative stress, and apoptosis in rats. Network pharmacology and Western blot results showed that the effect of EH extract on NS may be associated with the CAMKK2/AMPK/mTOR signaling pathway. Moreover, methylephedrine significantly ameliorated adriamycin-induced NRK-52e cell injury. Methylephedrine also significantly improved the phosphorylation of AMPK and mTOR, which were blocked by CC. In sum, EH extract may ameliorate renal injury via the CAMKK2/AMPK/mTOR signaling pathway. Moreover, methylephedrine may be one of the material bases of EH extract.
Rats ; Animals ; Doxorubicin/adverse effects* ; Nephrotic Syndrome ; AMP-Activated Protein Kinases/metabolism* ; Signal Transduction ; TOR Serine-Threonine Kinases/metabolism* ; Apoptosis

OBJECTIVE To establish the fingerprint of Zhuang medicine Jinmu granules and carry out chemometric analysis ， and determine the contents of three components . METHODS High performance liquid chromatography （HPLC） method was adopted. Using rutin as the reference ，HPLC fingerprints of 10 batches of Jinmu granules were drawn and similarity evaluation was performed by Similarity Evaluation of Chromatographic Fingerprint of Traditional Chinese Medicine （2012 edition）；the common peak was determined by comparing with mixed control ；SPSS 21.0 software was used for cluster analysis ，and SIMCA 14.1 software was used for principal component analysis and orthogonal partial least squares-discriminant analysis. The differential components affecting the quality of Jinmu granules were screened by taking the variable importance in projection （VIP）value ＞1 as the standard ；the HPLC method was used to determine the contents of astilbin ，polydatin and berberine hydrochloride in Jinmu granules. RESULTS There were 22 common peaks in 10 batches of Jinmu granules ，and the similarities were 0.962-0.997；five common peaks were identified ，namely gallic acid （peak 2），polydatin（peak 9），rutin（peak 11），astilbin（peak 13）and kaempferol （peak 20）. The results of cluster analysis showed that 10 batches of Jinmu granules could be clustered into 3 categories：S1 and S 3-S4 were grouped into one category ；S5-S6 and S 9 were grouped into one category ；S2，S7-S8 and S 10 were grouped into one category. The results of principal component analysis showed that the parameter of model interpretation was 0.951 and that of prediction ability was 0.723. The classification results were basically consistent with cluster analysis. The classifica tion results of orthogonal partial least squares- com discriminant analysis were also ba sically consistent with clus- ter analysis. The common peaks with VIP value ＞1 in the order were peak 7＞peak 11（rutin）＞peak 17＞peak 13（astilbin）＞ peak 3＞peak 8＞peak 6＞peak 16 respectively. The linear ranges of astilbin ，polydatin and berberine hydrochloride were 0.012 6- 1.225 0，0.010 8-1.052 5 and 0.020 0-1.562 5 mg/mL，respectively（all R 2＝0.999 9）. RSDs of precision ，stability（24 h）and repeatability tests were all less than 3％. The average recoveries were 99.48％（RSD＝2.67％，n＝9），98.57％（RSD＝1.77％，n＝ 9）and 100.84％（RSD＝2.49％，n＝9）. The contents were 1.221 0-7.011 6，2.251 1-4.462 9，1.252 4-3.328 7 mg/g，respectively. CONCLUSIONS Established fingerprint and the method of content determination are accurate ，stable and simple. Combined with chemometric analysis ，it can be used for the quality control and evaluation of Zhuang medicine Jinmu granules.

OBJECTIVE To prepar e and characterize danazol （DAZ）-sodium caseinate （SC）composite nanoparticles ，and to study the mechanism of preparing nanoparticles in “bottom-up”technology. METHODS SC was used as a stabilizer for regulating nanoparticles，so that DAZ-SC composite nanoparticles were prepared by anti-solvent precipitation method. The particle size ， Zeta-potential，micro-morphology，stability，encapsulation efficiency ，drug loading and in vitro dissolution rate were characterized. Fluorescence spectra ，IR spectra ，FBRM and other methods were used to analyze the interaction mechanism between DAZ and SC. RESULTS The particle size of DAZ-SC composite nanoparticles was （223.7±12.5）nm，and the polydispersity index was 0.274± 0.012. Zeta-potential was －（17.81±1.63）mV（n＝3）. The stability of nano-suspension was good ，the solid properties of DAZ were greatly improved ，and the dissolution rate was significantly increased. SC was statically quenched under the action of DAZ and the secondary structures of SC were changed. The crystallization process of DAZ was controlled under the action of SC ，and the interaction between DAZ and SC was mainly hydrogen bond and van der Waals force. CONCLUSIONS In this study ，DAZ-SC composite nanoparticles are successfully prepared. In the “bottom-up”technology，the interaction between SC and DAZ caused by hydrogen bond and van der Waals force inhibits the growth and agglomeration of drug crystals .