Objective To establish an ion chromatography method to determine the content of galactosamine in heparin sodium sample. Methods The content of galactosamine was determined by the ratio of response value of galactosamine and glucosamine.The determination was performed on an Dionex ICS, and the separation was carried out on a Amino acids capture column (30 mm ×3 mm), Series protect column (30 mm × 3 mm)and analytical column CarboPac PA20 (150 mm ×3 mm).The mobile phase was 14 mM potassium hydroxide solution at a flow rate of 0.4 mL/min; the column tempertature was at 30℃; the injection volume was 10μL.Results Glucosamine hydrochloride had good linearity within the range of 1.013 -16.211μg/mL(Y=2.303 4X+0.824 2,r=0.998 3), the average accuracy was 92.7%, and RSD was 3.2%(n=9), the limit of detection was 0.101 3μg/mL, and the limit of quantitation was 0.337 7μg/mL.D-Galactosamine hydrochloride had good linearity within the range of 0.010 2 -0.162 5 g/mL, (Y=31.157X-0.114 4,r=0.999 3).The accuracy was 102.1%, RSD was 2.4%(n=9).The limit of detection was 0.001 0μg/mL, and the limit of quantitation was 0.003 4μg/mL.The determination of galactosamine in 3 batches of heparin sodium raw material was not detected, (0.02 ±2.1)%, (0.03 ±1.5)%, respectively, which were all lower than the limit value (1%) of United States Pharmacopeia regulation.Conclusion The method for the determination of galactosamine in total hexose amine is successfully developed , which could be used as reference for improvement of the quality standard of heparin sodium.

OBJECTIVE To explore the effect of aqueous extract of Zheng Chaihu Yin(ZCH)on paracetamol(acetaminophen,APAP)-induced hepatotoxicity. METHODS Male ICR mice were divided into three scenarios randomly:the single treatment dose of ZCH,multiple treatment or pretreatment dose of ZCH. Each scenario had a up control group and an APAP model group,while single treatment dose of ZCH group had a ZCH group at the same time. The dose of APAP and ZCH was 500 mg·kg-1 and 36 g · kg- 1,respectively. 24 h after the last administration,plasma and liver samples were prepared. Ultra- performance liquid chromatography/quadrupole- time- of- flight mass spectrometry (UPLC-Q-TOF-MS)based metabolomics profiling was used to examine changes in plasma after expo?sure to ZCH,APAP or co-exposure to ZCH and APAP. Glutamic pyruvic transaminase (GPT) and glutamic oxaloacetic transaminas (GOT) values were determined by a biochemical auto analyzer in plasma. Histopathologic changes in the liver were observed and the area was calculated after HE staining. The data were analyzed with SPSS16.0 statistical software and the results were compared with the test between the two groups to find biomarkers. Also,SIMCA software was used for partial least squares-discriminant analysis (PLS-DA) pattern recognition. RESULTS Compared to control group, APAP dosing alone caused an increase in plasma transaminases and alterations in multiple metabolic pathways. Compared to APAP group,decrease in plasma transaminases was noted when ZCH was administered after or prior to APAP. Histopathologic results showed that in the single treatment group, multiple treatment group and pretreatment group,ZCH could alleviate the liver damage induced by APAP from (32.3 ± 12.0)% to (14.2 ± 9.9)%,(8.6 ± 7.9)% to (5.2 ± 1.7)% and (32.5 ± 10.0)% to (5.2 ± 6.4)%(P<0.05). Similarly,the PLS-DA of the LC-MS data showed that the groups dosed with APAP alone were the most distinct from controls,while animals dosed with ZCH prior to or after APAP treatment were located near control group. Metabolic spectrum results showed that ZCH could restore the changes in endogenous substances including lipid metabolism,amino acid metabolism,sugar metabolism and energy metabolism induced by APAP to normal. CONCLUSION ZCH water-extraction plays major roles in the regulation of metabolism on APAP-induced liver injury. These studies demonstrate that UPLC-Q-TOF-MS-based metabolomic analysis can be sensitively and accurately predict the initiation and progres?sion of liver injury and greatly contribute to a better understanding of the hepatoprotective effects of ZCH in a clinical environment.

Objective:To analyze the genetic stability of two recombinant rotaviruses (rLLR/NSP3 NLuc) and (rLLR/NSP3 CoV2/RBD) that are inserted and express exogenous genes for continuous passage and proliferation on MA104 cells.Methods:After measuring the titers of two recombinant rotavirus strains, they were transferred to the P2 generation according to MOI0.01. Subsequently, the previous generation of virus lysate was diluted and activated at 1∶100, and MA104 cells were continuously infected for 18 generations (P20). The virus titers of the P1, P5, P10, P15, and P20 generation of cell lysate were measured using indirect immunofluorescence, and RT-PCR identification and dsRNA PAGE silver staining were performed. The luciferase activity of rLLR/NSP3-NLUc was also detected.Results:No inserted fragment loss was found in the recombinant rotavirus rLLR/NSP3 NLuc within 20 generations, with recombinant virus titers ranging from 3.85~5.16 × 10 6 FFU/ml, with strong luciferase signals in each generation. The recombinant rotavirus rLLR/NSP3 CoV2/RBD showed loss of inserted fragments in the 6th generation, with infectivity titers ranging from 2.6 to 3.36 in the first 5 generations of the recombinant virus × 10 6 FFU/ml. Conclusions:The recombinant rotavirus with 582 bp NLuc inserted at the end of the NSP3 gene has good genetic stability, while the recombinant rotavirus with 885 bp RBD inserted at the end of the NSP3 gene was only stable in the first 5 generations, indicating that foreign genes can be inserted at the end of the NSP3 gene of the recombinant rotavirus, and the insert can express, but its stability requires more in-depth research.

Objective:To investigate the clinical and pathological features of congenital hepatic fibrosis (CHF).Methods:The clinical and pathological findings of 20 patients diagnosed with CHF from 2017 to 2023 were retrospectively analyzed.Results:Among the 20 patients, 8 were males and 12 were females with a median age of 21.5 years. Mostly patients were admitted to the hospital with cirrhosis, portal hypertension and upper gastrointestinal bleeding. Pathological features were diffuse fibrosis in the portal area, formation of fibrous septa of varying width, segmentation of the liver parenchyma, with hyperplasia of small bile ducts. Among them, 1 case (5%) was complicated with Caroli's disease, and 1 case (5%) was HNF1α hepatocellular adenoma. IHC GS showed that was positively expressed in acinar region 3 in 75% cases.Conclusion:CHF is mainly manifested by portal hypertension and its complications. Histopathology is the gold standard for diagnosis. The possibility of CHF should be considered first in children and adolescents with portal hypertension but no history of hepatitis, and complicated kidney disease. The positive pattern of acinus-3 region of GS in IHC is helpful for the diagnosis of CHF.

Objective To understand the infected strains and prevalence of brucellosis in occupational population in Tacheng and Kashgar regions,Xinjiang Uygur Autonomous Region.Methods In September 2015,blood samples from occupational population (including herders,semi-agricultural and semi-pastoral,veterinarians) and non-occupational population (including students and cadres) were collected to detect Brucellaspecific antibody and bacterial nucleic acids by rose bengal plate test (RBPT),serological standard tube agglutination test (SAT) and PCR methods,respectively.The positive products of PCR were sent to Shanghai Sangon Biotechnology Co.,LTD.Then the sequence results were retrieved online using the basic alignment search tool (BLAST) in GenBank web page and uploaded to NCBI (National Center for Biotechnology Information).Results A total of 546 blood samples were tested,including 300 males,aged (55-± 15) years old,and 246 females,aged (54 ± 12) years old.The positive rates were 17.58％ (96/546) and 6.78％ (37/546) in 546 blood samples by serological method and genetic markers targeting omp22 and omp2,respectively.The positive rates were statistically significant (x2 =29.8,P ＜ 0.05).Additionally,based on BLAST analysis of outer membrane protein omp22 and omp2 genes,the positive products were identified as Brucella abortus,and the sequence similarity was 100.00％ (253/253,863/863 bp) to Brucella abortus strain Wisconsin genome assembly,chromosome (LT651712).Conclusions Brucellosis has a high infection rate in the occupational population of some animal husbandry-based groups in Xinjiang.The infection strain is abortive species Brucella,and health education for the occupational population and prevention of brucellosis should be strengthened to reduce the infection rate.

Cystic echinococcosis (CE) caused by E. granulosus is a serious helminthic zoonosis in humans, livestock and wildlife. Xinjiang is one of high endemic province for CE in China. A total of 55 sheep and cattle livers containing echinococcal cysts were collected from slaughterhouses in Changji and Yining City, northern region of Xinjiang. PCR was employed for cloning 2 gene fragments, 12S rRNA and CO1 for analysis of phylogenetic diversity of E. granulosus. The results showed that all the samples collected were identified as G1 genotype of E. granulosus. Interestingly, YL5 and CJ75 strains were the older branches compared to those strains from France, Argentina, Australia. CO1 gene fragment showed 20 new genotype haploids and 5 new genotype haplogroups (H1-H5) by the analysis of Network 5.0 software, and the YLY17 strain was identified as the most ancestral haplotype. The major haplotypes, such as CJ75 and YL5 strains, showed identical to the isolates from Middle East. The international and domestic trade of livestock might contribute to the dispersal of different haplotypes for E. granulosus evolution.
Abattoirs ; Animals ; Argentina ; Australia ; Cattle ; China ; Clone Cells ; Cloning, Organism ; Echinococcosis ; Echinococcus granulosus ; Echinococcus ; France ; Genetic Variation ; Genotype ; Haploidy ; Haplotypes ; Helminths ; Humans ; Liver ; Livestock ; Middle East ; Polymerase Chain Reaction ; Sheep

Background::Triple-negative breast cancer (TNBC) is a type of highly invasive breast cancer with a poor prognosis. According to new research, long noncoding RNAs (lncRNAs) play a significant role in the progression of cancer. Although the role of lncRNAs in breast cancer has been well reported, few studies have focused on TNBC. This study aimed to explore the biological function and clinical significance of forkhead box C1 promoter upstream transcript (FOXCUT) in triple-negative breast cancer.Methods::Based on a bioinformatic analysis of the cancer genome atlas (TCGA) database, we detected that the lncRNA FOXCUT was overexpressed in TNBC tissues, which was further validated in an external cohort of tissues from the General Surgery Department of the First Affiliated Hospital of Nanjing Medical University. The functions of FOXCUT in proliferation, migration, and invasion were detected in vitro or in vivo. Luciferase assays and RNA immunoprecipitation (RIP) were performed to reveal that FOXCUT acted as a competitive endogenous RNA (ceRNA) for the microRNA miR-24-3p and consequently inhibited the degradation of p38. Results::lncRNA FOXCUT was markedly highly expressed in breast cancer, which was associated with poor prognosis in some cases. Knockdown of FOXCUT significantly inhibited cancer growth and metastasis in vitro or in vivo. Mechanistically, FOXCUT competitively bounded to miR-24-3p to prevent the degradation of p38, which might act as an oncogene in breast cancer. Conclusion::Collectively, this research revealed a novel FOXCUT/miR-24-3p/p38 axis that affected breast cancer progression and suggested that the lncRNA FOXCUT could be a diagnostic marker and therapeutic target for breast cancer.

With the increasing incidence and mortality of malignant tumors year by year， the prevention and treatment of malignant tumors has become a hot field of research. Although the existing anti-tumor treatment methods such as surgery， radiotherapy， chemotherapy， immunity and targeting have achieved some results， there are also obvious shortcomings， which are not conducive to the improvement of patients' quality of life and long-term treatment. Therefore， seeking a new efficient and safe treatment has become a hot topic of concern for many oncologists. adenosine monophosphate activated protein kinase（AMPK）/mammalian target of rapamycin（mTOR） signaling pathway can participate in intracellular energy metabolism and affect the pathogenesis and outcome of tumors through multiple mechanisms. It is widely mentioned in the occurrence and development of many tumors. The activity level of this pathway is closely related to the abnormal proliferation and invasion of tumor cells， and is considered as an important pathway for anti-tumor therapy research. The anti-tumor treatment of traditional Chinese medicine（TCM） has significant advantages such as syndrome differentiation and treatment and sample consideration. The study found that flavonoids， alkaloids， polyphenols， saponins and other active components of traditional Chinese medicine， as well as the traditional Chinese medicine compound Xiangbei San， Yiqi Fuzheng Jiedu decoction， Yiqi Fuzheng prescription， etc can activate the AMPK/ mTOR signaling pathway by regulating the expression of AMPK/mTOR signaling pathway and upstream and downstream proteins， thereby inducing tumor cell autophagy and apoptosis， inhibiting tumor angiogenesis and epithelial mesenchymal transformation， regulating aerobic glycolysis and energy metabolism， and reversing drug resistance to exert anti-tumor effects. In this paper， by summarizing the research achievements of TCM anti-tumor in recent years， the specific mechanism of action of TCM regulating AMPK/ mTOR signaling pathway against tumor was discussed， aiming to provide ideas and references for the research and development of anti-tumor new drugs.

Development of a vaccine that can simultaneously induce effective mucosal immunity and systemic immunity is an ideal goal to prevent mucosal pathogenic infections. The digestive tract has many sites for inducing mucosal immunity, including the mouth, stomach and small intestine. An ideal oral viral vaccine can not only induce better local and distal mucosal immunity, but also produce better systemic immunity. The oral viral vaccine has also attracted much attention because of its painless vaccination, self-administration and other advantages. Due to the complexity of human digestive tract environment and mucosal immunity, only three oral attenuated live vaccines have been successfully marketed for human use. This review summarizes the characteristics of gastrointestinal mucosal immunity, the current types and research status of oral viral vaccines, and the challenges faced by oral viral vaccines, with the hope to facilitate the research and development of oral viral vaccines for human use in China.
Humans ; Viral Vaccines ; Vaccination ; Immunity, Mucosal ; Vaccines, Attenuated ; Vaccine Development

:To investigate the effect of transient receptor potential melastatin 2 （TRPM2） inhibitor A10 on oxygen glucose deprivation/reperfusion （OGD/R） injury in SH-SY5Y cells．:Human neuroblastoma SH-SY5Y cells were subject to OGD/R injury，and then were divided into blank control group，model control group and A10 group randomly. The cell survival rate was detected by cell counting kit 8 （CCK-8）; the level of cellular reactive oxygen species （ROS） was detected by reactive oxygen detection kit; the mitochondrial membrane potential was detected by tetramethylrhodamine （TMRM） method; the number of apoptotic cells was detected by TUNEL apoptosis assay kit; the protein expression level of cleaved caspase 3 was detected by Western blot．:Compared with 3，20，30，50， has lower cytotoxicity and better inhibition effect on channel activity. Compared with the model control group，ROS level was reduced，the mitochondrial membrane potential was improved，the number of apoptosis cells was reduced ，and the expression of cleaved caspase 3 was significantly reduced in the A10 group（all <0.05）. : A10 can alleviate cell damage after OGD/R by inhibiting TRPM2 channel function，reducing extracellular calcium influx，reducing cell ROS levels，stabilizing mitochondrial membrane potential levels，and reducing apoptosis.
Apoptosis ; Benzeneacetamides ; Cell Survival ; Glucose ; Humans ; Oxygen/metabolism* ; Piperidones ; Reactive Oxygen Species/metabolism* ; Reperfusion ; TRPM Cation Channels