OBJECTIVE: To observe the clinical effect of needle cauterization on vitiligo with deficiency cold and blood stasis. METHODS: A total of 62 patients of vitiligo with deficiency cold and blood stasis were randomly divided into an observation group and a control group, 31 cases each group.On the basis of 308 nm excimer light irradiation combined with ironing with Chinese medicine, the control group was treated with tacrolimus ointment for external use, twice a day; the observation group was treated with needle cauterization at vitiligo spots, once a week.Both groups were treated for 10 weeks. Before and after treatment, the area of vitiligo spot, TCM syndrome score, serum levels of inflammatory indexes (interleukin[IL]-6 and IL-17) were observed in the two groups, and the clinical effect was evaluated. RESULTS: After treatment, the areas of vitiligo spot, TCM syndrome scores and serum levels of IL-6, IL-17 were decreased compared with those before treatment in both groups (P<0.05), and the above indexes in the observation group were lower than those in the control group (P<0.05). The total effective rate in the observation group was 93.5% (29/31), which was higher than 77.4% (24/31) in the control group (P<0.05). CONCLUSION Needle cauterization could reduce the areas of vitiligo spot in patients of vitiligo with deficiency cold and blood stasis, improve the clinical symptoms, its mechanism may be related to the reduction of serum levels of inflammatory indexes.
Humans ; Vitiligo/physiopathology* ; Male ; Female ; Adult ; Young Adult ; Middle Aged ; Adolescent ; Interleukin-6/blood* ; Interleukin-17/blood* ; Cautery ; Acupuncture Therapy/instrumentation* ; Needles ; Cold Temperature ; Treatment Outcome

Objective To investigate the serological and molecular characteristics of twenty blood samples carrying ABO?AW.37 allele and to analyze ABO allelic enhancement.Methods The ABO phenotype of the twenty samples was de-termined by serological methods and the genotype of 1-7 ABO exons was analyzed by Sanger sequencing.Results Sequen-cing analysis showed that all twenty samples contained a c.940A>G(p.Lys314Glu)mutation of A allele,which was defined as ABO?AW.37.When ABO?AW.37 and B alleles were inherited simultaneously in 9 cases,in forward typing anti-A anti-bodies all agglutinated and the serological phenotype was Aw B.Among the 11 cases with ABO?AW.37 and O alleles inherited simultaneously,there was no agglutination of anti-A in forward typing.For absorption and elution tests,5 cases were weakly positive and the serological phenotype was Ael,while 6 cases were negative for absorption and elution tests and the serologi-cal phenotype was O type.Conclusion Allelic enhancement occured when both ABO?AW.37 allele and B allele were in-herited simultaneously.When ABO? AW.37 was inherited simultaneously with O allele,the serological phenotype was Aelor O type and attention should be paid to blood type identification.

ObjectiveTo investigate the inhibitory effect of Mahuang Xixin Fuzitang on the migration of dendritic cells （DCs） in mice and its underlying mechanism. MethodMouse bone marrow-derived DCs were isolated and cultured. The morphological changes of the cells at different stages were observed under a microscope， and the CD11c⁺ proportion was detected by flow cytometry to identify DC purity. Cells were treated with Mahuang Xixin Fuzitang （1， 2， 5， 10， 20， 40， 50， 100 g·L^-1） for 24 hours， and the effect of Mahuang Xixin Fuzitang on cell proliferation was assessed using the cell counting kit-8 （CCK-8） assay to determine the appropriate concentrations for treatment. After modeling by lipopolysaccharide （LPS） induction， DCs were divided into a blank group， a model group， and Mahuang Xixin Fuzitang groups （2， 4， 8 g·L^-1）. The expression of surface molecules CD80， CD86， and major histocompatibility complex-Ⅱ （MHC-Ⅱ） were detected by flow cytometry. Transwell chamber assay was used to observe cell migration. The levels of chemokine C-C-primitive receptor 7 （CCR7） and chemokine C-X-C-primitive receptor 4 （CXCR4） on the cell surface were detected by enzyme-linked immunosorbent assay （ELISA）. The expression of filamentous actin （F-actin） in the cell microfilament cytoskeleton was detected by immunofluorescence （IF） staining. Real-time quantitative polymerase chain reaction （Real-time PCR） was employed to determine the mRNA expression levels of Ras homolog family member A （RhoA） and Rho-associated coiled-coil containing protein kinase 1 （ROCK1）. Western blot analysis was performed to detect the protein expression of RhoA and ROCK1. ResultCompared with the blank group， the model group exhibited significantly higher expression levels of CD80， CD86， and MHC-Ⅱ （P<0.01）， a significantly increased number of cells migrating to the lower chamber （P<0.01）， and significantly elevated levels of CCR7 and CXCR4 （P<0.05， P<0.01）. Additionally， F-actin expression was significantly increased （P<0.01）， and both RhoA and ROCK1 mRNA and protein expression levels were significantly higher （P<0.05， P<0.01）. Compared with the model group， treatment with Mahuang Xixin Fuzitang （2， 4， 8 g·L^-1） for 24 hours resulted in significantly lower expression levels of CD80， CD86， and MHC-Ⅱ （P<0.01）， a significantly reduced number of cells migrating to the lower chamber （P<0.05）， and significantly decreased levels of CCR7 and CXCR4 （P<0.05， P<0.01）. Furthermore， F-actin expression was significantly reduced （P<0.01）， and both RhoA and ROCK1 mRNA and protein expression levels were significantly decreased （P<0.05， P<0.01）. ConclusionMahuang Xixin Fuzitang can inhibit the migration of DCs in mice， and its mechanism of action may be related to reducing the activity of the Rho/ROCK signaling pathway， thereby affecting changes in the cell cytoskeleton.

ObjectiveTo investigate the effects of thyme herbal tea （BLX） on the proliferation of human coronavirus OC43 （HCoV-OC43） in vitro and in vivo. MethodThe chemical composition of BLX was analyzed by UPLC-MS. The cytotoxicity of BLX in HRT-18 cells and the effect of BLX treatment on the proliferation of HCoV-OC43 in cells were analyzed. Copies of viral gene were detected by real-time PCR. The effect of BLX treatment on the life cycle of HCoV-OC43 was detected by time-of-addition assay. The maximum tolerated dose of BLX and the influences of BLX on the body weight and survival time of suckling mice infected with HCoV-OC43 were determined. The expression of viral protein in the brain and lung tissue was analyzed by immunohistochemistry. ResultThere were 11 chemical components identified in BLX by UPLC-MS. BLX showed the 50% cytotoxic concentration （CC₅₀） of （13 859.56±319） mg·L^-1， the median inhibitory concentration （IC₅₀） of （1 439.09±200） mg·L^-1， and the selection index of 8.26-11.44 for HCoV-OC43 in HRT-18 cells. Compared with the cells infected with HCoV-OC43， BLX at the concentrations of 1 500， 1 000， 500 mg·L^-1 inhibited the proliferation of this virus （P<0.05， P<0.01）. BLX exhibited antiviral effect in the early stage of virus infection， and the inhibition role in the attachment stage was more significant than that in the entry stage （P<0.05）. In the suckling mice infected with HCoV-OC43， BLX at 1200 and 600 mg·kg^-1·d^-1 alleviated the symptoms， prolonged the survival period， reduced the death rate， and down-regulated the mRNA level of nucleocapsid protein in the mice. Moreover， BLX at 1 200 mg·kg^-1·d^-1 down-regulated the expression of nucleocapsid protein in the brain （P<0.01） and the lung （P<0.01）. ConclusionBLX contained multiple antiviral ingredients. It inhibited the proliferation of HCoV-OC43 both in vitro and in vivo by interference with viral attachment. This study provides theoretical reference for the treatment of acute respiratory tract infection with HCoV-OC43 and for further development and application of BLX.

Objective:To explore the relationship between anhedonia and suicidal ideation and the effects of anhedonia on suicidal ideation in patients with first-episode untreated depressive disorder.Methods:Totally 106 in-patients who met the diagnostic criteria for depressive disorder of the Diagnostic and Statistical Manual of Mental Disorders,Fifth Edition(DSM-5)were invited to participate in the study.Anhedonia,depressive symptoms and sui-cidal ideation were assessed with the Snaith-Hamilton Pleasure Scale(SHAPS),Hamilton Depression Scale(HAMD)and Beck Scale for Suicide Ideation(BSI),respectively.If both items 4 and 5 of BSI were 0,it indicated non-suicidal ideation,the subjects were divided into a group of suicidal ideation(n=66)and a group non-suicidal ideation(n=40).Spearman correlation analysis and multivariate binary logistic regression analysis were used for a-nalysis.Results:The correlation between the scores of SHAPS and HAMD was not statistically significant(P＞0.05).The SHAPS scores were positively correlated with the BSI scores(r=0.70,P＜0.01).Meanwhile,binary logistic regression analysis showed that anhedonia was one of a risk factor for suicidal ideation(OR=2.34,95％CI:1.58-3.47,P＜0.001).Conclusion:It suggests that anhedonia maybe partially independent of other depres-sive symptoms and it is one of the risk factors for suicidal ideation in first-episode untreated patients with depressive disorder.

Objective To construct recombinant human butyrylcholinesterase(rhBChE)knock-in goat fetal fibroblast cell lines(GFFs)by using clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated protein 9(Cas9)-mediated homology-directed repair mechanism for subsequent production of goat expressing rhBChE.Methods The efficient sgRNA sites targeting goat[3-casein(CSN2)gene were designed and screened,and the targeting efficiency of the sgRNA in goat mammary epithelial cells(GMECs)was confirmed by electro-transfection,flow sorting,and sequencing of PCR products.The red fluorescent reporter gene homology repair vector(P2A-mCherry)targeting the sgRNA was constructed,and then the integration and expression efficiency was detected by flow cytometry.The rhBChE homology repair vector(P2A-rhBChE)targeting the sgRNA of CSN2 gene was constructed in GFFs,the rhBChE positive cell clones were obtained via electro-transfection and flow sorting,and the rhBChE knock-in cell lines was identified by sequencing of PCR products.Results The sgRNA4 was identified as an efficient target of goat CSN2 gene,which could be also used for targeted integration of other genes.Three rhBChE knock-in cell lines were successfully constructed.Conclusion The rhBChE knock-in GFFs targeting goat CSN2 gene lays the foundation for the production of mammary bioreactors expressing rhBChE.

Objective To investigate the role and underlying mechanism of cathepsin B in myocar-dial injury in mice with diabetic cardiomyopathy(DCM).Methods Twenty 8-week-old male SPF C57BL/6 mice were randomly divided into wild-type(WT)group and WT DCM group,with 10 mice in each group.Another 20 8-week-old male SPF-grade mice with cathepsin B knockout(KO)were randomly and equally assigned to KO group and KO DCM group.HE staining was used to observe morphological changes,Prussian blue staining was employed to detect iron deposition,while immunohistochemical staining with 4-hydroxynonenal(4-HNE)was used to assess lipid peroxidation level in the myocardial tissues.Western blotting was performed to detect the expression of heme oxygenase-1(HO-1),superoxide dismutase 2(SOD2),and nuclear factor E2-related factor 2(Nrf2),while RT-PCR was applied to evaluate the expressions of Nrf-2,HO-1,and phospholipid hydroperoxide glutathione peroxidase 4(GPX4).Results Compared to the WT DCM group,the KO DCM group presented improved cell arrangement in cardiac tissues and sig-nificant reduction in inflammatory cell infiltration.Furthermore,the KO DCM group displayed a significant decrease in iron deposition compared to the WT DCM group.Additionally,the KO DCM group exhibited a significant reduction in 4-HNE expression compared to the WT DCM group.The protein levels of Nrf2,SOD2,and HO-1 were significant increased in the KO DCM group than the WT DCM group(0.68±0.21 vs 0.39±0.13,0.59±0.10 vs 0.28±0.09,1.03±0.10 vs 0.48±0.04,P＜0.05).Moreover,elevated mRNA levels of GPX4,Nrf2 and HO-1 were also observed in the KO DCM group than the WT DCM group(0.65±0.09 vs 0.40±0.10,0.61±0.11 vs 0.34±0.11,0.62±0.12 vs 0.39±0.09,P＜0.05).Conclusion Cathepsin B exacerbates myocardial injury in DCM mice through ferroptosis.

Mycoplasma capricolum subsp. capripneumoniae (Mccp) is the cause of contagious caprine pleuropneumonia (CCPP) in goats. Inactivated vaccines and capsular polysaccharide (CPS) indirect hemagglutination reagents are available for prevention and serological detection, but high culture costs and complex antigen quantification have been plagued by production staff. In order to solve these problems in production practice, a sugar fermentation medium with an initial pH value of 7.8, which could improve the production of two antigens simultaneously, was screened out by changing the initial pH value based on previous Mccp metabolomics analysis. Since phenol red can be identified by UV absorption spectrum and cetyltrimethylammonium bromide (CTAB) can bind to anionic capsular polysaccharide, a UV spectrum measurement method for analyzing the culture stage reached by Mccp and a CTAB precipitation test for relative quantification of capsular polysaccharide antigen content in the fermentation broth were established. The UV spectrum observation method can guide the production of Mccp according to the growth curve of Mccp, which greatly reduces the monitoring time of the traditional CCU method and improves the accuracy of the original eye-observation method. The established CTAB precipitation test can complete the monitoring of CPS content within 5 hours, which greatly reduces the time required compared with the traditional differential technique, and its accuracy was verified by the phenol-sulfuric acid method. The optimized culture medium and the two correlation comparison methods established in this study can effectively reduce the production cost of Mccp and improve the production efficiency. The two assays have been used in the research at our laboratory, which provides experimental data for further improvement of the production process of CCPP inactivated vaccine and capsular polysaccharide as well as rapid quantification.
Humans ; Animals ; Goats ; Cetrimonium ; Mycoplasma ; Polysaccharides

Objective:To investigate the therapeutic efficacy of venetoclax combined with avapritinib in treatment of refractory/relapsed acute myeloid leukemia (AML) with KIT gene mutation.Methods:The clinical data of 2 AML patients with KIT gene mutation who received venetoclax combined with avapritinib admitted to Canglang Hospital of Suzhou in October 2022 and November 2022 were retrospectively analyzed, and the relevant literature was reviewed.Results:Both patients with high-risk relapsed/refractory AML and KIT gene mutation were females; the one was 53 years and the other was 17 years. Case 1 was diagnosed with AML-M 2, and genetic testing revealed positive mutations in ASXL1, KIT, and RUNX1. The patient relapsed after transplantation and then was treated with venetoclax combined with avapritinib achieving morphologic leukemia-free status (MLFS). Case 2 was diagnosed with AML, and RUNX1-RUNX1T1 (AML1-ETO) fusion gene and KIT and DX15 gene mutations were detected. The patient was treated with venetoclax combined with avapritinib regimen after relapse, and the treatment regimen significantly reduced the tumor load. Complete remission was achieved after bridging to allogeneic hematopoietic stem cell transplantation. Conclusions:AML with KIT gene mutation is heterogeneous and some patients are difficult to treat with very poor prognosis. Bridging (secondary) hematopoietic stem cell transplantation can be the better treatment choice for relapsed patients achieving MLFS or complete remission after venetoclax combined with avapritinib treatment regimen.

The ABRACL protein, the regulator of actin and cell motility, belongs to the HSPC280 family, and its conserved hydrophobic groove can interact with other proteins to facilitate actin motility and cellular activity. ABRACL is upregulated in tumor tissues and is closely linked with the proliferation and migration of tumor cells. A deeper understanding of the role of ABRACL in tumorigenesis and development may provide new ideas and insights for ABRACL to prevent or reverse tumor progression.