Metachromatic leukodystrophy (MLD) is an inherited disease caused by a deficiency of the enzyme arylsulfatase A (ARSA). Lentivirus-modified autologous hematopoietic stem cell gene therapy (HSCGT) has recently been approved for clinical use in pre and early symptomatic children with MLD to increase ARSA activity. Unfortunately, this advanced therapy is not available for most patients with MLD who have progressed to more advanced symptomatic stages at diagnosis. Patients with late-onset juvenile MLD typically present with a slower neurological progression of symptoms and represent a significant burden to the economy and healthcare system, whereas those with early onset infantile MLD die within a few years of symptom onset. We conducted a pilot study to determine the safety and benefit of HSCGT in patients with postsymptomatic juvenile MLD and report preliminary results. The safety profile of HSCGT was favorable in this long-term follow-up over 9 years. The most common adverse events (AEs) within 2 months of HSCGT were related to busulfan conditioning, and all AEs resolved. No HSCGT-related AEs and no evidence of distorted hematopoietic differentiation during long-term follow-up for up to 9.6 years. Importantly, to date, patients have maintained remarkably improved ARSA activity with a stable disease state, including increased Functional Independence Measure (FIM) score and decreased magnetic resonance imaging (MRI) lesion score. This long-term follow-up pilot study suggests that HSCGT is safe and provides clinical benefit to patients with postsymptomatic juvenile MLD.
Humans ; Leukodystrophy, Metachromatic/genetics* ; Pilot Projects ; Genetic Therapy/methods* ; Hematopoietic Stem Cell Transplantation ; Male ; Follow-Up Studies ; Female ; Lentivirus/genetics* ; Child ; Child, Preschool ; Hematopoietic Stem Cells/metabolism* ; Cerebroside-Sulfatase/metabolism* ; Adolescent

Objective:To construct the sphingosine kinase 1(SPHK1)overexpression lentiviral vector,and to establish the SKOV3 lentiviral stable transfection cell line.Methods:According to the SPHK1 data information provided by the National Center for Biotechnology Information(NCBI)database,the primers were designed and synthesized,the target gene was amplified,and connected to the GV492 plasmid treated with Bam HⅠ and AgeⅠ restriction enzymes to construct the SPHK1 overexpression lentiviral vector;the positive clones were selected for PCR and sequencing identification;the lentiviral plasmid and the lentiviral packaging auxiliary plasmid were co-transfected into the HEK-293T cells for packaging and titer determination;according to the measured optimal multiplicity of infection(MOI)of 10,the corresponding lentiviral amounts in various groups were transfected into the SKOV3 cells,and the SKOV3 cells were divided into blank group(without treatment),GV492 control group(GV492 control lentivirus infected SKOV3 cells),and GV492-SPHK1 overexpression group(GV492-SPHK1 overexpression lentivirus infected SKOV3 cells,ov-SPHK1 group);the optimal concentration of 2 mg·L-1 puromycin was used to screen the stably transfected SKOV3 cell line;after 48 h,the medium was changed and replaced with 1 mg·L-1 puromycin for screening for 14 d;the morphology and fluorescence expression of the cells were observed under fluorescence microscope;real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of SPHK1 mRNA in the SKOV3 cells in various groups;Western blotting method was used to detect the expression level of SPHK1 protein in the SKOV3 cells in various groups.Results:The PCR sequencing results showed that the gene sequence of the SPHK1 overexpression lentiviral vector was completely consistent with the target sequence,and the SPHK1 overexpression lentiviral vector was successfully constructed;the titer determination results showed that the lentiviral titers in GV492 control group and ov-SPHK1 group were 5×1011 and 8×1011 TU·L?1,respectively;the SKOV3 cells in GV492 control group and ov-SPHK1 group were in good state and showed strong fluorescence expression,suggesting that the SKOV3 stable transfection cell line overexpressing SPHK1 was successfully established;the RT-qPCR results showed that compared with blank group and GV492 control group,the expression level of SPHK1 mRNA in the SKOV3 cells in ov-SPHK1 group was significantly increased(P<0.01);the Western blotting results showed that compared with blank group and GV492 control group,the expression level of SPHK1 protein in the SKOV3 cells in ov-SPHK1 group was significantly increased(P<0.01).Conclusion:The SPHK1 overexpression lentiviral vector is successfully constructed,and the SKOV3 stable transfection cell line is established.

This study was aimed at understanding the prevalence and influencing factors of food-borne parasitic diseases in ethnic minority areas of Guizhou Province,to provide a scientific basis for the development of appropriate intervention measures based on the human-animal-environment One Health concept.In 2023,the infection status of the human population,reservoir hosts,intermediate hosts,food-borne parasitic diseases,and related social and environmental factors were investigated in Congjiang County in Qidongnan Miao and Dong Autonomous Prefecture;Luodian County in Qiannan Buyi and Miao Autonomous Prefecture;and Ceheng County in Qianxinan Buyi and Miao Autonomous Prefecture.At least 1 000 individuals were sampled from each county,along with at least 50 insect-protected host samples from each location.Food-borne parasite infections were detected with the modified Kato thick smear method.A questionnaire survey was administered to the population.Detection of food-borne parasitic metacercariae was performed in intermediate host fish through the flaking and digestion method,and in crabs through the pounding and sedimentation method.The chi-square test was used to compare rates,and logistic regression was applied for multivariate analysis.A total of 3 023 questionnaires and fecal samples were collected.Males accounted for 47.50%,females accounted for 52.50%,and members of ethnic minorities accounted for 96.06%.A total of 186 food-borne parasitic infections were identified,and the infection rate was 6.15%.Five insect species were detected,which showed an infection rate of 5.39%.The infection rate of Clonorchis sinensis was 0.33%,that of Taenia was 0.40%,that of Heteroceles was 0.17%,that of Acanthus was 0.17%,and that of Echinostoma was 0.03%.Human infections with Echinostomus colloides and Echinostomia transferoris had not previously been reported in China.Single-factor analysis revealed statistically significant differences in food-borne parasite infections according to various factors,including the consumption of untreated water,raw fish and shrimp,raw pig blood,raw cow gastric juice,and raw pork and beef,as well as raw pig and cow viscera(P<0.05).Multivariate analysis indicated that the risk factors for food-borne parasite infections among residents in minority areas of Guizhou Province included the consumption of raw pig blood(OR=2.841,95%CI:1.809-4.463),raw cow gastric juice(OR=2.122,95%CI:1.297-3.469),and raw fish and shrimp(OR=1.779,95%CI:1.049-3.018).A total of 173 fecal samples of the reservoir host were examined,which showed a rate of food-borne parasite infection of 5.2%.A total of 510 intermediate host fish were examined,which showed a 4.51%positivity rate of encysted metacercaria of Clonorchis sinensis.The crab,pig,and beef samples were not positive.In conclusion,food-borne parasitic infections were prevalent in ethnic minority regions of Guizhou Province,and consumption of raw food were influencing factors.A focus on populations with raw food consumption habits,including raw pig blood,cow gastric juice,fish and shrimp,is essential.Concurrently,monitoring of animal hosts must be strengthened to perform key interventions according to the One Health concept.

This study was aimed at understanding the prevalence and influencing factors of food-borne parasitic diseases in ethnic minority areas of Guizhou Province,to provide a scientific basis for the development of appropriate intervention measures based on the human-animal-environment One Health concept.In 2023,the infection status of the human population,reservoir hosts,intermediate hosts,food-borne parasitic diseases,and related social and environmental factors were investigated in Congjiang County in Qidongnan Miao and Dong Autonomous Prefecture;Luodian County in Qiannan Buyi and Miao Autonomous Prefecture;and Ceheng County in Qianxinan Buyi and Miao Autonomous Prefecture.At least 1 000 individuals were sampled from each county,along with at least 50 insect-protected host samples from each location.Food-borne parasite infections were detected with the modified Kato thick smear method.A questionnaire survey was administered to the population.Detection of food-borne parasitic metacercariae was performed in intermediate host fish through the flaking and digestion method,and in crabs through the pounding and sedimentation method.The chi-square test was used to compare rates,and logistic regression was applied for multivariate analysis.A total of 3 023 questionnaires and fecal samples were collected.Males accounted for 47.50%,females accounted for 52.50%,and members of ethnic minorities accounted for 96.06%.A total of 186 food-borne parasitic infections were identified,and the infection rate was 6.15%.Five insect species were detected,which showed an infection rate of 5.39%.The infection rate of Clonorchis sinensis was 0.33%,that of Taenia was 0.40%,that of Heteroceles was 0.17%,that of Acanthus was 0.17%,and that of Echinostoma was 0.03%.Human infections with Echinostomus colloides and Echinostomia transferoris had not previously been reported in China.Single-factor analysis revealed statistically significant differences in food-borne parasite infections according to various factors,including the consumption of untreated water,raw fish and shrimp,raw pig blood,raw cow gastric juice,and raw pork and beef,as well as raw pig and cow viscera(P<0.05).Multivariate analysis indicated that the risk factors for food-borne parasite infections among residents in minority areas of Guizhou Province included the consumption of raw pig blood(OR=2.841,95%CI:1.809-4.463),raw cow gastric juice(OR=2.122,95%CI:1.297-3.469),and raw fish and shrimp(OR=1.779,95%CI:1.049-3.018).A total of 173 fecal samples of the reservoir host were examined,which showed a rate of food-borne parasite infection of 5.2%.A total of 510 intermediate host fish were examined,which showed a 4.51%positivity rate of encysted metacercaria of Clonorchis sinensis.The crab,pig,and beef samples were not positive.In conclusion,food-borne parasitic infections were prevalent in ethnic minority regions of Guizhou Province,and consumption of raw food were influencing factors.A focus on populations with raw food consumption habits,including raw pig blood,cow gastric juice,fish and shrimp,is essential.Concurrently,monitoring of animal hosts must be strengthened to perform key interventions according to the One Health concept.

Psoriasis is a refractory disease mainly co-acted by immune,genetic and environment.Tumor necrosis factor α (TNF-α)-related biologics have brought the landmark advances in the treatment of psoriasis;however,the anti-TNF-α therapy has the adverse response,its limitation may be related to the different bio-logical functions exerted by activation of TNF-α different receptors.Tumor necrosis factor receptor 2 (TNFR2) is one of the key receptors for TNF-α,and after binding to TNF-α,it can activate multiple signaling pathways such as NF-κB,PI3K/Akt,MAPK,STAT3,etc.,which are involved in the regulation of inflamma-tion,epidermal homeostasis,cellular apoptosis,cellular proliferation,cellular autophagy and other biological processes.It is suggested that TNFR2 is closely related to the occurrence and development of psoriasis.Previ-ous studies have often overlooked the role of TNFR2 in anti-TNF-α therapies;therefore,this article reviews the structure and signaling pathways of TNFR2,research advances in the disease,and its relationship with psoriasis to provide new references for exploring the pathogenesis and treatment of psoriasis.

Colorectal cancer (CRC) is a prevalent malignant tumor often leading to liver metastasis and mortality. Despite some success with PD-1/PD-L1 immunotherapy, the response rate for colon cancer patients remains relatively low. This is closely related to the immunosuppressive tumor microenvironment mediated by tumor-associated macrophages (TAMs). Our previous work identified that a phosphoglycerate mutase 1 (PGAM1) allosteric inhibitor, HKB99, exerts a range of anti-tumor activities in lung cancer. Here, we found that upregulation of PGAM1 correlates with increased levels of M2-like tumor-associated macrophages (TAMs) in human colon cancer samples, particularly in liver metastatic tissues. HKB99 suppressed tumor growth and metastasis in cell culture and syngeneic tumor models. M2-polarization, induced by colon cancer cell co-culture, was reversed by HKB99. Conversely, the increased migration of colon cancer cells by M2-TAMs was remarkably restrained by HKB99. Notably, a decrease in TAM infiltration was required for the HKB99-mediated anti-tumor effect, along with an increase in CD8⁺ T cell infiltration. Moreover, HKB99 improved the efficacy of anti-PD-1 treatment in syngeneic tumors. Overall, this study highlights HKB99's inhibitory activity in TAM-mediated colon cancer progression. Targeting PGAM1 could lead to novel therapeutic strategies and enhance the effectiveness of existing immunotherapies for colon cancer.

Objective:To compare the effectiveness and safety profile of centrifugal and membrane plasma separation model in artificial liver therapy with a dual plasma molecular adsorption system (DPMAS).Method:A retrospective study was conducted. Data of inpatients with liver failure who were treated with DPMAS therapy in the Liver Disease Center of Nanfang Hospital, Southern Medical University, from October 2022 to June 2024 were included. Clinical data such as demographic characteristics, etiology, DPMAS treatment-related indicators (including plasma separation method, vascular access, frequency of treatment, treatment duration, type of anticoagulant drugs, and membrane rupture condition), and laboratory test indicators before and after DPMAS treatment were collected. Categorical variables were compared by the χ2 test. Continuous variables were compared using a t-test or a non-parametric test between groups. Result:Data of 232 cases with liver failure who received artificial liver therapy with DPMAS were included. A total of 473 times DPMAS treatment was given. The average age was 50 years old, and males accounted for 82.3%. Centrifugal plasma separation was the initial DPMAS treatment in 176 (75.9%) cases, while membrane plasma separation was used in 56 cases (24.1%). The most common vascular access for DPMAS treatment was the internal jugular vein. The most commonly used anticoagulant was unfractionated heparin. The treatment duration of DPMAS was significantly higher with centrifugal separation than that with membrane separation ( P<0.001). Hemoglobin levels (mean before and after treatment in the centrifugal: 112.8 g/L vs. 106.3 g/L, P<0.001; mean before and after treatment in the membrane group: 108.4 g/L vs. 103.3 g/L, P<0.001), red blood cell count (mean before and after treatment in the centrifugal group: 3.7×10 9/L vs. 3.5×10 9/L, P<0.001; mean before and after treatment in the membrane group: 3.5×10 9/L vs. 3.3×10 9/L, P<0.001) and platelet count (mean before and after treatment in the centrifugal group: 134.5×10 9/L vs. 119.6×10 9/L, P<0.001; mean before and after treatment in the membrane group: 120.7 ×10 9/L vs. 97.3 ×10 9/L, P<0.001) were slightly decreased following initial DPMAS treatment in both groups. The decrease in platelets was significantly lower in centrifugal separation than that in membrane separation (median: 10.4% vs. 17.0%; P=0.003). There was no statistically significant difference observed in the proportion of puncture site bleeding in terms of plasma separation-related adverse events between the two groups, but plasma separator membrane rupture occurred two times in the DPMAS treatment. Conclusion:Centrifugal and membrane separation, both with DPMAS therapy, can cause a slight decrease in hemoglobin, red blood cell count, and platelets in patients with liver failure. Membrane separation causes a larger drop in platelets than centrifugal plasma separation. The operational convenience of medical personnel, the risk of membrane rupture, the coagulation markers, the patient's vascular condition, and other factors should be comprehensively considered when choosing the plasma separation model.

Objective:To compare the effectiveness and safety profile of centrifugal and membrane plasma separation model in artificial liver therapy with a dual plasma molecular adsorption system (DPMAS).Method:A retrospective study was conducted. Data of inpatients with liver failure who were treated with DPMAS therapy in the Liver Disease Center of Nanfang Hospital, Southern Medical University, from October 2022 to June 2024 were included. Clinical data such as demographic characteristics, etiology, DPMAS treatment-related indicators (including plasma separation method, vascular access, frequency of treatment, treatment duration, type of anticoagulant drugs, and membrane rupture condition), and laboratory test indicators before and after DPMAS treatment were collected. Categorical variables were compared by the χ2 test. Continuous variables were compared using a t-test or a non-parametric test between groups. Result:Data of 232 cases with liver failure who received artificial liver therapy with DPMAS were included. A total of 473 times DPMAS treatment was given. The average age was 50 years old, and males accounted for 82.3%. Centrifugal plasma separation was the initial DPMAS treatment in 176 (75.9%) cases, while membrane plasma separation was used in 56 cases (24.1%). The most common vascular access for DPMAS treatment was the internal jugular vein. The most commonly used anticoagulant was unfractionated heparin. The treatment duration of DPMAS was significantly higher with centrifugal separation than that with membrane separation ( P<0.001). Hemoglobin levels (mean before and after treatment in the centrifugal: 112.8 g/L vs. 106.3 g/L, P<0.001; mean before and after treatment in the membrane group: 108.4 g/L vs. 103.3 g/L, P<0.001), red blood cell count (mean before and after treatment in the centrifugal group: 3.7×10 9/L vs. 3.5×10 9/L, P<0.001; mean before and after treatment in the membrane group: 3.5×10 9/L vs. 3.3×10 9/L, P<0.001) and platelet count (mean before and after treatment in the centrifugal group: 134.5×10 9/L vs. 119.6×10 9/L, P<0.001; mean before and after treatment in the membrane group: 120.7 ×10 9/L vs. 97.3 ×10 9/L, P<0.001) were slightly decreased following initial DPMAS treatment in both groups. The decrease in platelets was significantly lower in centrifugal separation than that in membrane separation (median: 10.4% vs. 17.0%; P=0.003). There was no statistically significant difference observed in the proportion of puncture site bleeding in terms of plasma separation-related adverse events between the two groups, but plasma separator membrane rupture occurred two times in the DPMAS treatment. Conclusion:Centrifugal and membrane separation, both with DPMAS therapy, can cause a slight decrease in hemoglobin, red blood cell count, and platelets in patients with liver failure. Membrane separation causes a larger drop in platelets than centrifugal plasma separation. The operational convenience of medical personnel, the risk of membrane rupture, the coagulation markers, the patient's vascular condition, and other factors should be comprehensively considered when choosing the plasma separation model.

Objective To analyze the genetic subtypes of human immunodeficiency virus (HIV) and the prevalence of pretreatment drug resistance in the newly reported HIV-infected men in Guangxi. Methods The stratified random sampling method was employed to select the newly reported HIV-infected men aged≥50 years old in 14 cities of Guangxi from January to June in 2020.The pol gene of HIV-1 was amplified by nested reverse transcription polymerase chain reaction and then sequenced.The mutation sites associated with drug resistance and the degree of drug resistance were then analyzed. Results A total of 615 HIV-infected men were included in the study.The genetic subtypes of CRF01_AE,CRF07_BC,and CRF08_BC accounted for 57.4% (353/615),17.1% (105/615),and 22.4% (138/615),respectively.The mutations associated with the resistance to nucleoside reverse transcriptase inhibitors (NRTI),non-nucleoside reverse transcriptase inhibitors (NNRTI),and protease inhibitors occurred in 8 (1.3%),18 (2.9%),and 0 patients,respectively.M184V (0.7%) and K103N (1.8%) were the mutations with the highest occurrence rates for the resistance to NRTIs and NNRTIs,respectively.Twenty-two (3.6%) patients were resistant to at least one type of inhibitors.Specifically,4 (0.7%),14 (2.3%),4 (0.7%),and 0 patients were resistant to NRTIs,NNRTIs,both NRTIs and NNRTIs,and protease inhibitors,respectively.The pretreatment resistance to NNRTIs had much higher frequency than that to NRTIs (2.9% vs.1.3%;χ²=3.929,P=0.047).The prevalence of pretreatment resistance to lamivudine,zidovudine,tenofovir,abacavir,rilpivirine,efavirenz,nevirapine,and lopinavir/ritonavir was 0.8%, 0.3%, 0.7%, 1.0%, 1.3%, 2.8%, 2.9%, and 0, respectively. Conclusions CRF01_AE,CRF07_BC,and CRF08_BC are the three major strains of HIV-infected men≥50 years old newly reported in Guangxi,2020,and the pretreatment drug resistance demonstrates low prevalence.
Male ; Humans ; Middle Aged ; Reverse Transcriptase Inhibitors/therapeutic use* ; HIV Infections/drug therapy* ; Drug Resistance, Viral/genetics* ; China/epidemiology* ; Mutation ; HIV-1/genetics* ; Protease Inhibitors/therapeutic use* ; Genotype