OBJECTIVE To establish a quality control method for Jinbei oral liquid based on multi-wavelength switching high performance liquid chromatography （HPLC） fingerprint， chemical pattern recognition and multi-index content determination. METHODS A total of 15 batches of Jinbei oral liquid were used as test samples. The Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine （ Version 2012 ） was adopted to establish multi-wavelength switching HPLC fingerprints， followed by chromatographic peak identification and similarity evaluation. Cluster analysis， principal component analysis and orthogonal partial least squares-discriminant analysis were applied to conduct chemical pattern recognition on the 15 batches of samples. The multi-wavelength switching HPLC method was used to simultaneously determine the contents of neochlorogenic acid， chlorogenic acid， cryptochlorogenic acid， forsythoside A， baicalin， salvianoli c acid B and wogonoside in samples. RESULTS A total of 25 common peaks were calibrated in the fingerprints of 15 batches of Jinbei oral liquid， among which 7 common peaks were unambiguously identified. The similarity of all samples was higher than 0.960. Chemical pattern recognition results showed that samples S1-S8 were clustered into group 1， and samples S9-S15 were clustered into group 2. Baicalin， wogonoside， neochlorogenic acid， chlorogenic acid and salvianolic acid B were identified as the differential quality markers. The average contents of neochlorogenic acid， chlorogenic acid， cryptochlorogenic acid， forsythoside A， baicalin， salvianolic acid B and wogonoside in 15 batches of Jinbei oral liquid were 0.213 7， 0.085 3， 0.104 9， 0.287 3， 0.420 1， 0.062 1， 0.176 0 mg/mL， respectively. CONCLUSIONS The established quality control method for Jinbei oral liquid combining multi-wavelength switching HPLC fingerprint， chemical pattern recognition and multi-index content determination is stable and reliable， which can provide a reference for the formulation of quality standards of this preparation.

OBJECTIVE To establish a quality control method for Jinbei oral liquid based on multi-wavelength switching high performance liquid chromatography （HPLC） fingerprint， chemical pattern recognition and multi-index content determination. METHODS A total of 15 batches of Jinbei oral liquid were used as test samples. The Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine （ Version 2012 ） was adopted to establish multi-wavelength switching HPLC fingerprints， followed by chromatographic peak identification and similarity evaluation. Cluster analysis， principal component analysis and orthogonal partial least squares-discriminant analysis were applied to conduct chemical pattern recognition on the 15 batches of samples. The multi-wavelength switching HPLC method was used to simultaneously determine the contents of neochlorogenic acid， chlorogenic acid， cryptochlorogenic acid， forsythoside A， baicalin， salvianoli c acid B and wogonoside in samples. RESULTS A total of 25 common peaks were calibrated in the fingerprints of 15 batches of Jinbei oral liquid， among which 7 common peaks were unambiguously identified. The similarity of all samples was higher than 0.960. Chemical pattern recognition results showed that samples S1-S8 were clustered into group 1， and samples S9-S15 were clustered into group 2. Baicalin， wogonoside， neochlorogenic acid， chlorogenic acid and salvianolic acid B were identified as the differential quality markers. The average contents of neochlorogenic acid， chlorogenic acid， cryptochlorogenic acid， forsythoside A， baicalin， salvianolic acid B and wogonoside in 15 batches of Jinbei oral liquid were 0.213 7， 0.085 3， 0.104 9， 0.287 3， 0.420 1， 0.062 1， 0.176 0 mg/mL， respectively. CONCLUSIONS The established quality control method for Jinbei oral liquid combining multi-wavelength switching HPLC fingerprint， chemical pattern recognition and multi-index content determination is stable and reliable， which can provide a reference for the formulation of quality standards of this preparation.

OBJECTIVES: To explore the active components that mediate the therapeutic effect of Centella asiatica on psoriasis and their therapeutic mechanisms. METHODS: TCMSP, TCMIP, PharmMapper, Swiss Target Prediction, GeneCards, OMIM and TTD databases were searched for the compounds in Centella asiatica and their targets and the disease targets of psoriasis. A drug-active component-target network and the protein-protein interaction network were constructed, and DAVID database was used for pathway enrichment analysis. In a RAW264.7 macrophage model of LPS-induced inflammation, the anti-inflammatory effect of 7.5, 15, 30, and 60 μmol/L quercetin, asiaticoside, and asiatic acid, which were identified as the main active components in Centella asiatica, were tested by measuring cellular production of NO, TNF‑α and IL-6 using Griess method and ELISA and by detecting mRNA expressions of IL-23, IL-17A, TNF-α and IL-6 and protein expressions of p-STAT3 (Tyr705) and p-STAT3 (Ser727) with RT-qPCR and Western blotting. RESULTS: A total of 139 targets of Centella asiatica and 4604 targets of psoriasis were obtained, and among them CASP3, EGFR, PTGS2, and ESR1 were identified as the core targets. KEGG analysis suggested that quercetin, asiaticoside, and asiatic acid in Centella asiatica were involved in cancer and IL-17 and MAPK signaling pathways. In the RAW264.7 macrophage model of inflammation, treatment with quercetin significantly reduced cellular production of NO, TNF‑α and IL-6, and lowered mRNA expressions of IL-23, IL-17A, TNF‑α and IL-6 and protein expressions of p-STAT3 (Tyr705) and p-STAT3 (Ser727). CONCLUSIONS Quercetin, asiaticoside and asiatic acid are the main active components in Centella asiatica to mediate the therapeutic effect against psoriasis, and quercetin in particular is capable of suppressing cellular production of NO, TNF‑α and IL-6 and regulating the IL-23/IL-17A inflammatory axis by mediating STAT3 phosphorylation to inhibit inflammatory response.
Quercetin/pharmacology* ; Psoriasis/metabolism* ; STAT3 Transcription Factor/metabolism* ; Mice ; Animals ; Centella/chemistry* ; Triterpenes/pharmacology* ; Phosphorylation ; Interleukin-17/metabolism* ; Interleukin-23/metabolism* ; RAW 264.7 Cells ; Pentacyclic Triterpenes/pharmacology* ; Macrophages/drug effects* ; Signal Transduction ; Plant Extracts

Objective:To investigate the G/P genotypes of group A rotavirus（RVA）in the 2023 sentinel surveillance in Jiangsu Province，and to conduct a molecular characterization analysis of the whole-genome sequences of four G9P［4］genotype RVA strains identified during surveillance.Methods:A total of 212 RVA-positive specimens collected from the surveillance system in 2023 were subjected to G/P genotyping using multiplex nested RT-PCR. Whole-genome sequencing was performed on six G9P［4］strains. The resulting complete genome sequences were preliminarily genotyped using BLAST，followed by comprehensive molecular characterization analyses utilizing BioEdit 7.0.5，MAFFT，MEGA 7.0，and iTOL software.Results:The overall RVA positivity rate was 6.22%. The predominant G/P combination in both outpatient and inpatient settings was G8P［8］. Among the six G9P［4］strains，four were successfully sequenced. All four exhibited the genotype constellation G9-P［4］-I2-R2-C2-M2-A2-N1-T2-E2-H2. While the NSP2 gene belonged to the N1 genotype，all other genes corresponded to the DS-1-like genogroup. Phylogenetically，the four Jiangsu G9P［4］strains clustered within Lineage V of the VP7 gene and formed a distinct minor subclade within the N1 branch of the NSP2 gene. Unique amino acid substitutions were identified at multiple VP7 neutralization antigenic epitope sites when compared to vaccine strains.Conclusions:The predominant circulating RVA strain in Jiangsu province during 2023 was G8P［8］. Concurrently，the relatively uncommon G9P［4］-N1 strain was detected. This strain exhibited significant amino acid differences at key epitopes compared to vaccine strains. Enhancing the proportion of whole-genome sequencing in RVA surveillance is warranted to obtain more detailed genetic information，thereby providing crucial data to support future vaccine development and optimization strategies.

Objective:To investigate the G/P genotypes of group A rotavirus（RVA）in the 2023 sentinel surveillance in Jiangsu Province，and to conduct a molecular characterization analysis of the whole-genome sequences of four G9P［4］genotype RVA strains identified during surveillance.Methods:A total of 212 RVA-positive specimens collected from the surveillance system in 2023 were subjected to G/P genotyping using multiplex nested RT-PCR. Whole-genome sequencing was performed on six G9P［4］strains. The resulting complete genome sequences were preliminarily genotyped using BLAST，followed by comprehensive molecular characterization analyses utilizing BioEdit 7.0.5，MAFFT，MEGA 7.0，and iTOL software.Results:The overall RVA positivity rate was 6.22%. The predominant G/P combination in both outpatient and inpatient settings was G8P［8］. Among the six G9P［4］strains，four were successfully sequenced. All four exhibited the genotype constellation G9-P［4］-I2-R2-C2-M2-A2-N1-T2-E2-H2. While the NSP2 gene belonged to the N1 genotype，all other genes corresponded to the DS-1-like genogroup. Phylogenetically，the four Jiangsu G9P［4］strains clustered within Lineage V of the VP7 gene and formed a distinct minor subclade within the N1 branch of the NSP2 gene. Unique amino acid substitutions were identified at multiple VP7 neutralization antigenic epitope sites when compared to vaccine strains.Conclusions:The predominant circulating RVA strain in Jiangsu province during 2023 was G8P［8］. Concurrently，the relatively uncommon G9P［4］-N1 strain was detected. This strain exhibited significant amino acid differences at key epitopes compared to vaccine strains. Enhancing the proportion of whole-genome sequencing in RVA surveillance is warranted to obtain more detailed genetic information，thereby providing crucial data to support future vaccine development and optimization strategies.

Objective To explore the approach to establish a temporal lobe epilepsy model via intraperitoneal pilocarpine injection in C57BL/6J mice,and to summarize behavioral indicators predicting successful modeling during the acute phase of epileptic seizures after pilocarpine administration,aiming to offer a practical mice model for future epilepsy research.Methods Thirty C57BL/6J substrain mice(primary subjects)and forty C57BL/6N substrain mice(control subjects)were selected to establish a temporal lobe epilepsy model by inducing seizures through a single intraperitoneal injection of pilocarpine.The mice from the two substrains were each divided into 3 groups,and were injected intraperitoneally with 300 mg/kg,330 mg/kg,or 360 mg/kg of pilocarpine,respectively.Motor seizure behaviors were observed and compared between the two substrains of C57BL/6 mice post pilocarpine injection,and the spontaneous recurrent seizures(SRS)were continuously monitored from the 7th day after injection.On the 28th day post-injection,the mice were euthanized and the histopathological changes in their hippocampi were examined.Results After pilocarpine administration,C57BL/6N mice displayed characteristic motor seizures followed by the onset of status epilepticus(SE).Conversely,C57BL/6J mice showed fewer instances of typical motor seizure behavior and the subsequent SE.Instead,they more often exhibited systemic tremors lasting several seconds to tens of seconds following limb twitching.This behavior is classified as"severe seizure(SS)"in current study.Following intraperitoneal injection of 330 mg/kg and 360 mg/kg pilocarpine,C57BL/6J mice displaying SS during the acute phase of seizure might exhibit SRS after a latency period.The percentage of spontaneous seizures observed in C57BL/6J mice post-modeling(70% )was comparable to that seen in C57BL/6N mice(75% )which developed SRS subsequent to SE.C57BL/6J mice displayed characteristic pathological alterations associated with temporal lobe epilepsy in the hippocampi after 28 d following pilocarpine injection,including increased mossy fiber sprouting and neuronal death.Conclusions When inducing an epilepsy model via intraperitoneal pilocarpine injection in C57BL/6J mice,the behavioral criteria to predict the successful establishment of the model could be either the occurrence of SE or the manifestation of SS.

Objective:To evaluate the efficacy of allogeneic hematopoietic cell transplantation(allo-HSCT) using unrelated cord blood or haploidentical donors in the treatment of children with primary immunodeficiency diseases (PID).Methods:The clinical data of 60 children with PID admitted to Chinese People′s Liberation Army General Hospital-Sixth Medical Center from April 2014 to October 2019 were retrospectively analyzed, including 56 cases of chronic granulomatous disease, 2 cases of severe combined immunodeficiency disease, 1 case of high-IgM syndrome and 1 case of severe congenital neutropenia.All patients underwent allo-HSCT, including 12 cases receiving the transplantation from unrelated cord blood (UCB group) and 48 cases from haploidentical donors combined with a third party unrelated cord blood (haploid group). Among these patients, there were 59 males and 1 female, with a median age of 3.4 years.All patients received a myeloablative conditioning regimen based on Busulfan.The prophylaxis of acute graft versus host disease (aGVHD) was performed based on Cyclosporine.In the UCB group, the median dose of mononuclear cells and CD 34+ cells was 0.67×10 8/kg and 0.51×10 6/kg recipient body weight, respectively; In the haploid group, bone marrow and peripheral stem cells from haploid donors were infused on day 01 and day 02, respectively.The third party cord blood was infused 4 hours before bone marrow infusion.The median dose of mononuclear cells and CD 34+ cells of bone marrow and peripheral stem cells from haploid donors was 9.97×10 8/kg and 5.12×10 6/kg recipient body weight, respectively.Kaplan-Meier method was used to analyze the overall survival rate. Results:The median day to neutrophil and platelet engraftment was 13.0 days and 23.5 days, respectively.The rate of complete donor chime-rism was shown 30.0 days after transplantation.There was no case with primary engraftment failure, and 1 case with secondary engraftment failure.The incidence of grade Ⅰ-Ⅱ and grade Ⅲ-Ⅳ aGVHD was 43.3% and 15.5%, respectively.The incidence of chronic graft versus host disease with limited skin type was 6.7%, while that with extensive type was 1.1%.The median follow-up period was 818 days.There were 6 death cases, among which, 5 cases died from infection and 1 case died from heart failure.The total mortality related to transplantation was 11.9%.A total of 53 cases survived without diseases.The estimated 5-year failure free survival and overall survival rate was 83.9% and 88.1%, respectively.Conclusion:The efficacy of allo-HSCT in the treatment of children with PID using unrelated cord blood and haploidentical donors is favorable.

Porcine reproductive and respiratory syndrome (PRRS) is a highly contagious disease caused by porcine reproductive and respiratory syndrome virus (PRRSV), which causes great economic losses. At the moment, no effective neutralizing antibody is available for scientific research and treatment. Therefore, developing a method for screening the neutralizing monoclonal antibodies is of great significance for the prevention and treatment of PRRSV and the screening of antigen sites. Monoclonal antibodies have been widely used in the treatment and diagnosis of many human and animal diseases. Therefore, screening effective neutralizing antibodies for different pathogens is an urgent task. Among the methods for monoclonal antibody screening, B cell immortalization is an effective method to obtain neutralizing monoclonal antibody. Specifically, in this study, the bcl-6 and bcl-xl genes were connected by f2a and then the yielded product was ligated to a vector for retrovirus packaging. The swine lymphocytes immunized with PRRSV were infected the yielded mature viruses and cultured in the complete medium containing CD40L and IL21 cytokines. Then, CD21 was used as the marker to screen B cells with the magnetic bead method. Finally, monoclonal B cells were obtained and the secretion of antibodies was tested. The results showed that the plasmid, either being transfected alone or with the packaged plasmids, could be expressed, and that the packaged retrovirus could infect the cells. Moreover, the infected lymphocytes secreted antibodies, so did the screened B cells. Therefore, the method for screening monoclonal antibody against PRRSV was successfully established.
Animals ; Antibodies, Monoclonal ; Antibodies, Neutralizing ; Antibodies, Viral ; Humans ; Porcine Reproductive and Respiratory Syndrome/prevention & control* ; Porcine respiratory and reproductive syndrome virus/genetics* ; Swine

Objective:To explore the efficacy of haploidentical hematopoietic stem cell transplantation (haplo-HSC) combined with tpCB in the treatment of children with bone marrow failure syndromes (BMFs).Methods:The clinical chara-cteristics of 78 BMFs pediatric patients, including inherited BMFs (4 cases) and acquired BMFs (74 cases) under-went haplo-HSC combined with the third-party cord blood (tpCB) in Chinese People′s Liberation Army General Hospital-Sixth Medical Center between July 2012 and July 2019 and were retrospectively analyzed, with 41 males and 37 females.Among them, 73 cases experienced first transplantation and 5 cases accepted second transplantation, with the median age of 5.6 years.The conditioning regimen was based on Busulfan, with 74 acquired BMFs cases using non-myeloablative and the remaining 4 cases using myeloablative.The prophylaxis of acute graft versus host disease (aGVHD) includes Cyclosporine, Mycophenolate mofetil (MMF) and Methotrexate.All patients received bone marrow from haploid donor and tpCB on day 1 and peripheral stem cell from haploid donor on day 2.The median dose of the total donor nucleated cells was 12.19×10 8/kg of recipient weight and CD 34+ cell dose was 6.13×10 6/kg of recipient weight. Results:The median time of granulocytes over 0.5×10 9/L and platelets over 20×10 9/L were + 13 d and + 17 d, respectively.All patients displayed complete donor-type chimerism at + 30 d. No primary graft failure occurred in any patient and second graft failure occurred in two cases.The incidence rate of grade ⅡtoⅣ and grade Ⅲ to Ⅳ aGVHD were 39.0% and 13.9%, respectively.The incidence of chronic GVHD with limited type and extensive type were 7.8% (95% CI: 7.1%-8.5%) and 2.6% (95% CI: 2.1%-3.1%), respectively.the median follow-up was 1 550 days, and 76 patients survived with free disease.The rate of transplant related mortality was 2.8%, and both of the estimated 5-year overall survival and failure-free survival rate were 97.2%(95% CI: 96.8%-97.6%). Conclusions:Haplo-HSC and umbilical cord blood can quickly provide hematopoietic stem cells.The results of haplo-HSC combined with the tpCB in pediatric patients with life-threatening BMFs are promising.

Objective: To understand the knowledge, attitude and behavior of HIV/AIDS prevention and treatment of freshmen in Jiangsu Colleges, so as to provide evidence for carrying out HIV/AIDS prevention and treatment propaganda among young college students. Methods: A total of 2 538 freshmen in 8 colleges in Jiangsu province were surveyed by cluster sampling, and anonymous self-reported questionnaire survey was carried out. Valid questionnaires were collected and analyzed. Results: Among 2 538 students valid, the total awareness rate of the eight items of new HIV/AIDS basic knowledge which provided to the public by the State (referred to the new eight items) was 85.74%, the awareness rate of boys was higher than that of girls’(88.21%, 84.34%)(P<0.05), the rate of college students of science was higher that of those of arts(87.31%, 83.67%)(P<0.05). the general awareness rate of other knowledge was 72.14%, and the rate of boys was higher than that of girls(75.00%, 70.53%)(P<0.05). As for "whether eating with the AIDS-affected patients may be affected with AIDS/HIV", boys and students of science were more rational(58.3%, 55.1%; 50.4%, 50.9%)(P<0.05). 50.08% students approved of having sexual behavior before marriage, and the rate of boys was higher than that of girls(69.00%, 39.40%)(P<0.01). 71.24% of the students thought that condoms should be used consistently in sex behaviors, and the proportion of female students (80.76%) was much higher than that of male students(54.37%), and the rate of the students of science was higher than that of students of arts(73.36%, 69.63%)(P<0.05). 6.03% students had received consult and test of AIDS. 40.34% of the untested students were willing to receive the test but didn’t know where to go. 1.34% students were afraid of receiving the test because of fear. The most popular ways of HIV/AIDS publicity among students are micro films(76.32%), lectures(66.67%) and peer education(60.80%). Conclusion It is suggested to publicize HIV/AIDS knowledge among students according to different gender and subject characteristics and combining multimedia, with emphasis on strengthening the publicity of HIV/AIDS epidemiology,treatment and related information, and attaching importance to sexual health education among young students.