OBJECTIVE To establish a high-performance liquid chromatography （HPLC） fingerprint and content determination method for Phellodendron chinensis ， and to evaluate its quality combined with chemometrics analysis.METHODS The HPLC fingerprints of 30 batches of P. chinensis were established by the Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine （2012 Edition）， and similarity evaluation and common peak identification were carried out.The contents of phellodendrine hydrochloride， magnoflorine， chlorogenic acid， 5- O -feruloylquinic acid， 4- O -feruloylquinic acid， 3- O -feruloylquinic acid， jatrorrhizine hydrochloride，and berberine hydrochloride were determined by the same HPLC method. Hierarchical cluster analysis （HCA）， principal component analysis （PCA）， and partial least squares-discriminant analysis （PLS-DA） were performed using the contents of the above eight components as variables.RESULTS A total of 16 common peaks were identified in the HPLC fingerprints of 30 batches of samples， and 8 common peaks were identified， including phellodendrine hydrochloride， magnoflorine， chlorogenic acid， 5- O -feruloylquinic acid， 4- O -feruloylquinic acid， 3- O -feruloylquinic acid， jatrorrhizine hydrochloride，and berberine hydrochloride. The similarity of 30 batches of samples ranged from 0.989 to 1.000. The results of content determination showed that the contents of eight components such as phellodendrine hydrochloride were 10.108-22.745，2.598-5.529，6.250-13.942，4.956-12.433，10.895-27.384，43.248-78.924， 0.428-1.154， 81.827-153.066 mg/g， respectively. The HCA results showed that 30 batches of samples were clustered into 3 categories：S1-S10 as one category， S11-S23 as one category，and S24-S30 as one category. The PCA results showed that the comprehensive scores of 10 batches of samples from the Sichuan production area （S1-S10） ranked among the top 10. The PLS-DA results showed that the variable importance in projection values of berberine hydrochloride， 3- O -feruloylquinic acid， chlorogenic acid and phellodendrine hydrochloride were greater than 1.CONCLUSIONS The HPLC fingerprint and content determination methods established in this study are stable and reliable. Chemometric analysis based on multi-component content data can be used for the quality evaluation of P. chinensis . Berberine hydrochloride， phellodendrine hydrochloride，and 3- O -feruloylquinic acid may be the differential markers affecting the overall quality of P. chinensis .

Objective To analyze the short-term clinical efficacy and prognosis of one-stop transcatheter aortic valve replacement (TAVR)+percutaneous coronary intervention (PCI) in the treatment of aortic valve disease with coronary heart disease. Methods The clinical data of patients with aortic valve disease complicated with coronary heart disease who underwent one-stop TAVR+PCI treatment at the Department of Cardiovascular Surgery, the Second Hospital of Hebei Medical University from January 2018 to June 2023 were retrospective analyzed. The preoperative and postoperative clinical data were compared, and 1-month follow-up results were recorded. Results A total of 37 patients were enrolled, including 22 males and 15 females, with an average age of 69.14±6.47 years. Thirty-six patients recovered and were discharged after the surgery, and 1 (2.7%) patient died during the surgery. Self-expanding TAVR valves were implanted through the femoral artery in all patients. One coronary artery was opened by PCI in 35 (94.6%) patients, and two coronary arteries were opened by PCI in 2 (5.4%) patients. All PCI opened arteries had a stenosis>70%. During the postoperative hospitalization, the complications included pulmonary infection in 11 (30.6%) patients, severe pneumonia in 10 (27.8%) patients, liver function injury in 14 (38.9%) patients, renal function injury in 5 (13.9%) patients, cerebral infarction in 1 (2.8%) patient, atrial fibrillation in 1 (2.8%) patient, ventricular premature beats in 2 (5.6%) patients, atrioventricular block in 2 (5.6%) patients, and complete left bundle branch block in 5 (13.9%) patients. The median postoperative ventilation assistance time was 12.0 (0.0, 17.0) h, the ICU monitoring time was 1.0 (0.0, 2.0) d, and the postoperative hospitalization time was 5.0 (4.0, 7.0) d. There was a significant improvement in the New York Heart Association cardiac function grading after surgery (P<0.001). After surgery, there were 21 (58.3%) patients had minor perivalve leakage, 6 (16.7%) patients had minor to moderate perivalve leakage, and no moderate or above degree of perivalve leakage. After one month of postoperative follow-up, 36 patients showed significant improvement in heart function. There were no patients with recurrent acute coronary syndrome, re-PCI, or cardiovascular system disease related re-hospitalization. Conclusion The one-stop TAVR+PCI treatment for patients with aortic valve disease and coronary heart disease can obtain satisfactory short-term clinical efficacy, which is worth further trying and studying.

OBJECTIVE To explore the clinical features and surgical treatment methods of primary parapharyngeal space tumors.METHODS The clinical data of 38 patients with primary parapharyngeal space tumors who were treated in the Affiliated Hospital of Southwest Medical University from June 2005 to April 2023 were retrospectively analyzed,and their clinical manifestations,imaging data,surgical methods,postoperative complications,postoperative pathological types and prognosis were analyzed.RESULTS The clinical manifestations of 38 patients were mainly pharyngeal discomfort and pharyngeal distension.CT and MRI can determine the size,location,and relationship of the tumor to surrounding tissue.The surgical methods included transcervical approach in 17 cases,transoral approach in 15 cases,transcervical-transparotid approach in 4 cases,and transcervical-transmandibular approach in 2 cases.Postoperative pathology diagnosed 28 cases(73.7%)of benign tumors and 10 cases(26.3%)of malignant tumors.After the postoperative follow-up of 2 months-17 years,there was no recurrence of benign tumor patients,and patients with malignant tumors survived well after comprehensive treatment with surgery and radiotherapy and chemotherapy.CONCLUSION Most patients with primary parapharyngeal space tumors have no specific clinical symptoms,and the types of pathologies are diverse.Choosing the appropriate surgical approach to remove the tumor is key to treatment.Neurologic deficits are the most important postoperative complication.Patients with benign tumors have good long-term effects after surgery,and patients with malignant tumors have better efficacy in the combination of surgery and chemoradiotherapy.

Anti-seizure medications (ASMs) are the main therapy for epilepsy.There are many kinds of ASMs with complex mechanism of action, so it is difficult for pharmacists to examine prescriptions.This paper put forward some suggestions on the indications, dosage forms/routes of administration, appropriateness of usage and dosage, combined medication and drug interaction, long-term prescription review, individual differences in pathophysiology of children, and drug selection when complicated with common epilepsy, for the reference of doctors and pharmacists.

Recombinant HLA-Ⅰ molecules/antigenic peptide complexes (pHLA complexes) are applied in the research of human T cell-specific immune responses. The preparation of pHLA complex is based on genetic engineering and protein in vitro dilution and folding-refolding technology. In an in vitro refolding system, recombinant HLA-Ⅰ molecules correctly fold and bind with antigenic peptides to form complexes. In this study, ultrafiltration-high performance liquid chromatography (ultrafiltration-HPLC) was used for quantitative determination of the antigenic peptides in recombinant pHLA complexes, especially for those in a small amount of prepared products. By adding the recombinant HLA-Ⅰ molecules and antigenic peptides into the refolding buffer, the heavy chain (HC) and light chain (β2m) of recombinant HLA-Ⅰ molecules were refolded and bond with the VYF antigenic peptide containing anchor residues to form a pHLA complex. The unbound free antigenic peptide VYF was removed by ultrafiltration to retain the complex. Finally, the pHLA complex was treated by acid to destroy its interaction, thus releasing the antigenic peptide. The results showed that the prepared recombinant pHLA complex was recognized by HLA-Ⅰ molecule specific antibody W6/32, which indicated that the recombinant HLA-Ⅰ class molecule had correct folding and was identified as pHLA complex. The antigen peptide VYF contained in the pHLA complex was also detected by ultrafiltration-HPLC, so it is feasible to apply ultrafiltration-HPLC for determination of pHLA complex. Compared with Western blotting, the concentration of antigenic peptides detected by ultrafiltration-HPLC was 0-9 μg/mL. The binding conditions can be optimized according to the amount of antigenic peptides bound in the complex in order to improve the folding efficiency of HLA-Ⅰ molecules and promote the binding of HLA-Ⅰ molecules to antigenic peptides. The production rate of pHLA complexes in the refolding system can also be calculated according to the content of antigenic peptides bound by pHLA complexes. Therefore, ultrafiltration-HPLC in this study can be used for the quality control of the preparation process of pHLA complexes, and may facilitate the research of T cell-specific immunity, artificial antigen-presenting cells, and development of specific tetramer probe applications.
Amino Acid Sequence ; Antigens ; Chromatography, High Pressure Liquid ; Humans ; Peptides/chemistry* ; Ultrafiltration

Objective:To explore the relationship between high performance liquid chromatography (HPLC) fingerprint and toxicity of Gelsemium elegans fermented by Ganoderma lucidum ( Ganoderma lucidum- Gelsemium elegans) for different time. Methods:Gelsemium elegans was processed by biphasic solid-state fermentation with Ganoderma lucidum. A total of 10 samples of Ganoderma lucidum- Gelsemium elegans were collected after fermentation for 9, 11, 13, 15, 17, 19, 21, 23, 25, 27 days (sampling twice on day 27, number: S1-S10) and the fingerprints were determined by self-established HPLC. One hundred specific pathogen-free ICR mice were randomly divided into 10 groups (each group comprised 10 mice, half were male and half were female). The median lethal dose of Ganoderma lucidum- Gelsemium elegans collected after fermentation for 11 days in mice was used as the final concentration in toxicity test. S1-S10 Ganoderma lucidum- Gelsemium elegans solutions were prepared and given to 10 groups of mice respectively by gavage administration and the death of mice was observed. According to the calculation formula of grey correlation analysis, the correlation coefficients between the common peaks of S1-S10 Ganoderma lucidum- Gelsemium elegans in chromatographic fingerprint and their toxicity test results (death rate in mice) was calculated and the main components contributing to the toxicity of Ganoderma lucidum- Gelsemium elegans were analyzed. Results:A total of 17 common peaks were identified in the chromatographic fingerprint spectrum of S1-S10 Ganoderma lucidum- Gelsemium elegans. The mortalities in mice caused by S1-S10 of Ganoderma lucidum- Gelsemium elegans were 1.00, 1.00, 0.80, 0.70, 0.60, 0.60, 0.50, 0.40, 0, and 0, respectively. The grey correlation analysis showed that the correlation coefficients of common peak 7, 3, 6, 9, 1, 8, 17, and 12 to toxicity were 0.868, 0.838, 0.830, 0.828, 0.824, 0.820, 0.818, and 0.802, respectively. The chemical components represented by these 8 chromatographic peaks had more contribution to the toxicity of Ganoderma lucidum- Gelsemium elegans. Conclusions:With the extension of fermentation time, the toxicity of Ganoderma lucidum- Gelsemium elegans decreased gradually, and toxicity was the lowest at 27 days of fermentation. The toxicity of Gelsemium elegans after fermentation was the result of a join action from multiple components. The identification of the main toxicity components can provide a reference for the quality control of Ganoderma lucidum- Gelsemium elegans and the fermentation process optimization.

Objective:To explore the relationship between high performance liquid chromatography (HPLC) fingerprint and toxicity of Gelsemium elegans fermented by Ganoderma lucidum ( Ganoderma lucidum- Gelsemium elegans) for different time. Methods:Gelsemium elegans was processed by biphasic solid-state fermentation with Ganoderma lucidum. A total of 10 samples of Ganoderma lucidum- Gelsemium elegans were collected after fermentation for 9, 11, 13, 15, 17, 19, 21, 23, 25, 27 days (sampling twice on day 27, number: S1-S10) and the fingerprints were determined by self-established HPLC. One hundred specific pathogen-free ICR mice were randomly divided into 10 groups (each group comprised 10 mice, half were male and half were female). The median lethal dose of Ganoderma lucidum- Gelsemium elegans collected after fermentation for 11 days in mice was used as the final concentration in toxicity test. S1-S10 Ganoderma lucidum- Gelsemium elegans solutions were prepared and given to 10 groups of mice respectively by gavage administration and the death of mice was observed. According to the calculation formula of grey correlation analysis, the correlation coefficients between the common peaks of S1-S10 Ganoderma lucidum- Gelsemium elegans in chromatographic fingerprint and their toxicity test results (death rate in mice) was calculated and the main components contributing to the toxicity of Ganoderma lucidum- Gelsemium elegans were analyzed. Results:A total of 17 common peaks were identified in the chromatographic fingerprint spectrum of S1-S10 Ganoderma lucidum- Gelsemium elegans. The mortalities in mice caused by S1-S10 of Ganoderma lucidum- Gelsemium elegans were 1.00, 1.00, 0.80, 0.70, 0.60, 0.60, 0.50, 0.40, 0, and 0, respectively. The grey correlation analysis showed that the correlation coefficients of common peak 7, 3, 6, 9, 1, 8, 17, and 12 to toxicity were 0.868, 0.838, 0.830, 0.828, 0.824, 0.820, 0.818, and 0.802, respectively. The chemical components represented by these 8 chromatographic peaks had more contribution to the toxicity of Ganoderma lucidum- Gelsemium elegans. Conclusions:With the extension of fermentation time, the toxicity of Ganoderma lucidum- Gelsemium elegans decreased gradually, and toxicity was the lowest at 27 days of fermentation. The toxicity of Gelsemium elegans after fermentation was the result of a join action from multiple components. The identification of the main toxicity components can provide a reference for the quality control of Ganoderma lucidum- Gelsemium elegans and the fermentation process optimization.

For multicellular organisms, cell-cell communication is essential to numerous biological processes. Drawing upon the latest development of single-cell RNA-sequencing (scRNA-seq), high-resolution transcriptomic data have deepened our understanding of cellular phenotype heterogeneity and composition of complex tissues, which enables systematic cell-cell communication studies at a single-cell level. We first summarize a common workflow of cell-cell communication study using scRNA-seq data, which often includes data preparation, construction of communication networks, and result validation. Two common strategies taken to uncover cell-cell communications are reviewed, e.g., physically vicinal structure-based and ligand-receptor interaction-based one. To conclude, challenges and current applications of cell-cell communication studies at a single-cell resolution are discussed in details and future perspectives are proposed.
Animals ; Cell Communication ; Humans ; RNA-Seq ; Single-Cell Analysis ; Transcriptome

Objective:To investigate the availability, prices and affordability of essential medicines in pediatric population across China, in the hope of improving rational use of medicines.Methods:A multicenter cross-sectional survey of medicine prices, availability and affordability was conducted in 17 provinces, municipalities and autonomous region across east, south-central part, west and north of China. Data on 42 medicines used in pediatric population, both original and generic, were collected in 55 public hospitals from May 26 to June 2, 2017. Availability was expressed as the percentage of hospitals with stock of the target medicine on the day of data collection,and median price ratio (MPR) was the ratio of price upon investigation to international reference. Based on national minimum daily wage, affordability represents the number of working days needed to earn the expense which covers a standard course using the target medicine. Statistical software SPSS 13.0 was applied for descriptive analysis of availability, MPR and affordability.Results:Mean Availability of original and generic medicine was 33% and 32%, with median MPR being 5.43 and 1.55. Among the 19 medicines with price information for both original and generic product, the median MPR was 7.73 and 2.04 respectively. Regarding the five medicines used to treat four common pediatric diseases (pneumonia,peptic ulcer, congenital hypothyroidism, refractory nephrotic syndrome), the affordability was 0.63 (0.16-6.17) d for generic medicine, and 1.03 (0.16-11.53) d for its original counterpart.Conclusions:The availability to both original and generic products of the 42 medicines used in pediatric population was low in China. The prices of generic medicines seem to be lower and affordability higher than those of original medicines. There is an urgent need to improve the availability and affordability of pediatric medicines.

Objective To explore the effect of functional electrical stimulation combined with treadmill training and botulinum toxin type A injection on foot-drop and strephenopodia among stroke survivors.Methods Sixty-seven stroke survivors with foot-drop and strephenopodia were randomly divided into an electrical stimulation group (n=23),a conventional treatment group (n =22) and a combined treatment group (n =22).All 3 groups received a 400 U injection of BTX-A and electrical stimulation.After 24 hours,the patients in the conventional treatment group received conventional treatment including a brain protection agent,limb function exercises,gait training,balance training and training in the activities of daily living.The patients in the combined treatment group received that conventional treatment,plus functional electrical stimulation and weight loss training on a treadmill.The patients in the electrical stimulation group received functional electrical stimulation treatment supplementing the conventional treatment.Therapeutic effects were evaluated before and after six weeks of treatment using integral electromyography (iEMS) of the anterior tibial muscle and the lateral head of the gastrocnemius muscle,the co-contraction ratio (CR) during ankle dorsiflexion,the modified Ashworth Scale (MAS),the Berg balance scale (BBS),a functional walking score (FAC),and the active range of motion (AROM) of the ankle in dorsiflexion and eversion.Results After the treatment,significant improvement was observed in all three groups in the average iEMS value of the anterior tibial muscle and the lateral head of the gastrocnemius muscle,the CR in ankle dorsiflexion,and in their MAS,BBS,FAC and AROM results.There was no significant difference among the three groups after treatment in their average iEMS values at the lateral head of the gastrocnemius.The average values of the other indicators were,however,significantly better in the combined treatment group than in the other 2 groups.Conclusion Functional electrical stimulation combined with treadmill training and botulinum toxin type A injection can significantly improve foot-drop,strephenopodia and the walking function of stroke survivors.This combined treatment deserves popularization and application in clinical practice.