OBJECTIVE:To establish a RP-HPLC method for simultaneous determination of chlorapheniramine maleate,furacilin and ephedrine hydrochloride in Puma nose drops.METHODS:The analysis was carried on a XDB C 8 column;the mo?bile phase was composed of methanol(A),acetonitrile(B)and0.02mol/L potassium dihydrogen phosphate solutions(containing0.2%triethylamine and adjusted to pH3.0with phosphoric acid,C)with linear gradient elution(0min～3.5min,A∶B∶C=6∶13∶18,8.5min,A∶B∶C=6∶30∶64)and the flow rate was1.0ml/min;the detection wavelength was254nm and the column temperature was30℃.Chloramphenicol was used as the internal standard.RESULTS:The linear ranges were0.04～0.20mg/ml for chlorapheniramine maleate,0.02～0.10mg/ml for furacilin,0.50～2.50mg/ml for ephedrine hydrochlo?ride.The average recoveries were99.44%(RSD=0.48%,n=3)for chlorapheniramine maleate,101.36%(RSD=0.41%,n=3)for furacilin and99.43%(RSD=0.59%,n=3)for ephedrine hydrochloride.CONCLUSION:The method is reliable,accurate and suitable for quality control of Puma nose drops.

AIM:To improve accuracy of the diagnosis of primary conjunctival masses(PCM) through analyze the clinicopathological characteristics and pathogenic causes of 422 cases of PCM.METHODS:Clinical manifestations, histological characteristics and immunopheno types were reviewed.Relevant literatures were reviewed.RESULTS:Benign conjunctival masses were 403 cases (95.5%).Those included masses from squamous cell papilloma (71 cases, 17.6%), conjunctival cyst (68 cases, 16.9%), inflammatory masses (62 cases, 15.4%), pigmented nevus (60 cases, 14.9%), lipoma (58 cases, 14.4%).Malignant conjunctival neoplasms were 19 cases (4.5%).Those included tumors from B Cell Lymphoma (6 cases,31.6%), melanoma (6 cases,31.6%), and squamous cell carcinoma (5 cases, 26.3%).Different types of primary conjunctival masses were induced by the same cause.CONCLUSION:The pathological types of primary conjunctival masses are various and the majority of which is benign.For the diagnosis of primary conjunctival masses, morphological analysis, immunohistochemical examination shoud be combined to increase the diagnostic accuracy and decrease misdiagnosis rate.

Mitochondrial dysfunction has been implicated in the aetiology of sporadic Parkinson's disease but its role in the disease mechanism remains unclear.To investigate the effect of synuclein on mitochondrial dysfunction induced by rotenone.The human dopaminergic SH-SY5Y cells were used as a cell model.The cells over-expressed the wild-type ?-synuclein were treated with complex I inhibitor rotenone.The cell viability,complex I activity,Mitochondrial swelling and O2-content were tested at different time point-1w,2w,4w after rotenone treated.CCK-8 test results showed that the cell viability of overexpressed ?-synuclein(SH-SY5Y-Syn)was much lower than the control group(SH-SY5Y-Ctr).After administrating with rotenone about 1w or 2w the cell viability of SH-SY5Y-Syn became higher than that of SH-SY5Y-Ctr.On the 4th week the results were contrary to the first 2 weeks.Similar results were got when test the mitochondrial function.In the first 2 weeks after roteoone administrating,the mitochondrial function of SH-SY5Y-Syn was better than that of SH-SY5Y-Ctr.This suggest that the ?-synuclein could protect the mitochondrial against the injury induced by rotenone in the early stage-1w,2w,while this effect disappeared in the final stage-4w.

Objective:To identify the effect of ?-synuclein overexpression on mitochondrial membrane structure with atomic force microscopy. Methods:?-syn expression was mediated by AAV (adeno-associated viral vector) and Recombinant AAV/?-syn and AAV/LacZ viral particles were stereotaxically injected in the left side of rat substantia nigra (SN) for rat model of ?-synuclein overexpression. Mitochondria were isolated from rats SN of Brain. Mitochondria were analysis with JC-1 staining,atomic force microscopy and Western blot. Results:By 16 weeks post-infection of AAV-?-syn,the level of ?-syn increased about 2 times in mitochondrial fraction with Western blot and mitochondrial membrane potential (??) decreased with JC-1 staining. Furthermore,mitochondria swelling and porous like structure formed on the mitochondrial membrane with atomic force microscopy. Conclusion:The data suggested that ?-syn could accumulate in mitochondria,might form mitochondrial membrane pores and lead to ?? decreases. ?-syn might lead to mitochondrial dysfunction in Parkinson's disease.

OBJECTIVE The aim of the present study was to investigate the anti-tumor effect and mechanism of a novel compound, C3C12PPD, a bioactive unnatural ginsenoside by metabolically engi-neered yeasts based on a new UDP-glycosyl- transferase from Bacillus subtilis. METHODS MTT assay was used to analyze the anti-proliferation activity of C3C12PPD in vitro. The effect of anti-tumor activity was observed by mouse Lewis xenograft model in vivo.The effects of C3C12PPD on suppressing the angio-genesis and invasion of A549 cells were investigated in vitro using Transwell and tube formation assays. RNAseq was used to find tagets of C3C12PPD. Western blotting was performed to investigate the expres-sion level of proteins in tumor tissues treated with C3C12PPD. RESULTS C3C12PPD could inhibit the growth of lung cancer in vitro and in viv o. At the dosage of 10.0 mg·kg-1, C3C12PPD inhibited tumor growth by 40.0% (P<0.05) in tumor weight in mouse Lewis xenograft. The inhibition of tube formation was 77.5%(P<0.01)and 80.3%(P<0.01)following treatment with 1×10-4and 2×10-4mol·L-1C3C12PPD for 5 h, whereas the proliferation of EA.hy926 cells was not influenced under the above concentrations. Under the concentrations of 1×10-4mol·L-1,C3C12PPD inhibited invasive ability of A549 cells(P<0.05).The results of RNAseq susgested that antitumor activity of C3C12PPD were associated with epithelial-mesenchymal transition (EMT) and angiogenesis. Moreover, the proteins related to EMT, Raf/MEK/ERK and AKT/mTOR signal pathways were effected by C3C12PPD analysed by western blotting. CONCLUSION These data suggested that C3C12PPD was able to supress lung cancer through inhibit EMT, invision and angiogenesis.

Abstract: Objective　To analyze the clinical features of hemophagocytic syndrome (HPS) associated with Orientia tsutsugamushi disease in children. Methods　The case data of patients with scrub typhus in Kunming Children's Hospital from January 1st 2019 to December 31st 2021 was retrospectively analyzed. The patients were divided into the HPS group and the non-HPS group according to whether associated with HPS. The clinical data of the two groups were analyzed using SPSS 25.0. Results　Eighty-five cases of scrub typhus in children were collected, 15 cases (17.6%) had HPS. The mean age of patients with HPS was (5.10±3.82) years, included 9 males and 6 female, there was no significant difference in gender and age between the HPS and the non-HPS group (P>0.05). Comparison of the two groups indicted that the incidence of cough, lung rales, edema, and hepatomegaly were significantly increased in the HPS group (P<0.05). The data showed that compared to the non-HPS group, the HPS group showed significant decreases in the levels of hemoglobin (HGB), platelet (PLT), albumin (ALB), fibrinogen (Fib) (P<0.05), and significant decreases in the levels of C-reactive protein (CRP), lactic dehydrogenase (LDH), triglyceride (TG), serum ferritin (SF) (P<0.05). The proportion of CD4+ T lymphocytes, CD4+/CD8+ were significantly decreased (P<0.05); the proportion of CD3+, CD8+ T lymphocytes were significantly increased (P<0.05). The proportion of pulmonary exudation or consolidation in the HPS group was higher than the non-HPS group, which was statistically significant (P<0.05). All the patients with scrub typhus associated with HPS were treated with oral doxycycline, and intravenous immunoglobulin was given in 13 cases (86.7%). There was one case of death and 14 cases discharged from hospital after treatment in HPS group. Conclusion　HPS in scrub typhus infected children is a nonnegligible complication. Prolonged fever, lung rales, hepatomegaly，HGB decreased, thrombocytopenia, hyperferritinemia, and abnormal lymphocyte subsets may associate with HPS. It should be alerted to scrub typhus when presenting with HPS in endemic areas. The scrub typhus associated with HPS can be successfully treated with appropriate antibiotic and immunomodulator treatment.

Animals ; Animals, Newborn ; Astragalus Plant ; Doxorubicin ; adverse effects ; Drugs, Chinese Herbal ; pharmacology ; Mice ; Mice, Inbred BALB C ; Myocardium ; pathology ; ultrastructure

In this study, 31 Notopterygium incisum populations were analyzed using ITS sequences to investigate the genetic structure. The results showed that: the ITS region ranged in size from 634 to 635 bp and base composition was with high G + C content of 57.8%. Thirty-one polymorphic sites were detected from 402 sequences of 31 populations of N. incisum, and the proportion of polymorphic sites was 4.88%, in which parsimony informative sites were up to 12. And 31 haplotypes were identified based on these polymorphic sites. Molecular variance analysis (AMOVA) indicated that high genetic differentiation (57%) existed among population, and gene flow was low (N(m) = 0.38) among populations. Phylogenetic relationships of 31 haplotypes were analyzed using NJ method with N. forbesiias an out-group. Phylogenetic analysis showed that 31 haplotypes from different populations mixed together and did not form distinct geographically separated clades.
Apiaceae ; classification ; genetics ; Base Sequence ; China ; DNA, Intergenic ; genetics ; Gene Flow ; Genetic Variation ; Molecular Sequence Data ; Phylogeny

Objectives To analyze the diagnosis and management of steroid-resistant nephrotic syndrome (SRNS) accompanied with irreversible leukoencephalopathy in children. Methods The clinical, laboratory and imaging data were retrospectively analyzed in a SRNS child accompanied with irreversible leukoencephalopathy. A literature review was performed. Results After clinical diagnosis of SRNS, glucocorticoid, immunosuppressant, and hemodialysis were administrated for 10 months. During the course of treatment, the seizures, visual problems, and hypertension were repeatedly occured. The cranial MRI showed bilateral occipital parietal lobe hyperintensity and right frontotemporal lobe hyperintensity on T2-weighted imaging and bilateral occipital parietal lobe hypointensity on T2-Flair imaging, which indicated that encephalomalacia was accompanied with gliosis. Conclusions A variety of reasons may induce leukoencephalopathy in children. The accompanied irreversible leu-koencephalopathy should be strongly considered in management of SRNS.

Objective:To clone and express Mycobacterium tuberculosis(Mtb) Rv2460c gene(encoding ClpP2 protein),and evaluate the immunogenicity of its coding product. Methods: The recombinant plasmid of pET32a (+) vector-ClpP2 that H37Rv Rv2460c gene was cloned into the plasmid pET32a(+)vector,was transformed into E. coli BL21(DE3)and induced expression by IPTG,then purified by affinity chromatography. The recombinant protein was confirmed by SDS-PAGE and Western blot. The analysis of immunogenicity of Mtb ClpP2 and its epitope prediction were performed by bioinformatic methods. The antibody levels of polyclonal antibody titer against ClpP2 protein in rabbits and TB patients′ serum were detected by ELISA. Results: The recombinant ClpP2 protease was expressed as inclusion bodies in E. coli. The purity of purified protein was 93% by bandscan software analysis. The bioin-formatics analysis shows Mtb ClpP2 protein has multiple preponderant B cell and T cell epitopes. Rabbit antiserum titer was 1∶64 000;Serum anti-ClpP2 antibody levels in TB patients was higher than that in healthy control subjects. Conclusion:The recombinant ClpP2 protein was purified, and specific Rabbit anti-ClpP2 polyclonal antibody was prepared successfully. Experiment and bioinformatic information studies showed that Mtb ClpP2 protease has strong immunogenicity.