Objective:To establish ferroptosis-related risk characteristics, to evaluate the prognostic correlation of ferroptosis-related genes in hepatocellular carcinoma, and to explore the complex relationship between hepatocellular carcinoma, ferroptosis and immune microenvironment.Methods:The bioinformatics analysis involved obtaining ferroptosis-related differentially expressed genes (DEGs) from the GeneCards database and the cancer genome atlas database. The biological functions of ferroptosis-related DEGs were analyzed using gene ontology and Kyoto encyclopedia of genes and genomes pathway enrichment. Ferroptosis-related DEGs clusters were identified using univariate Cox regression analysis and cluster analysis, etc. The correlation between ferroptosis-related DEGs clusters and tumor immune microenvironment and tumor occurrence score was evaluated using immunopanoramic analysis and tumor-related score analysis. Based on ferroptosis-related characteristics, a ferroptosis-related characteristic spectrum and nomogram were constructed using multivariate Cox regression and correlation analysis, etc. The correlation between the risk characteristics and tumor immune microenvironment, tumor occurrence score and gene mutation were evaluated using immune panoramic analysis, tumor-related score analysis and gene mutation analysis. In the experimental verification stage, the mRNA expression levels of aurora kinase A ( Aurka), acetyl-CoA carboxylase alpha ( Acaca) and arrestin domain containing 3 ( Arrdc3) in mouse primary hepatocytes and mouse hepatoma Hepa1-6 cells were verified by real-time reverse transcription-PCR (RT-qPCR). The mRNA expression levels of AURKA, ACACA and ARRDC3 in adjacent normal tissues and tumor tissues of patients with hepatocellular carcinoma were verified by RT-qPCR. A heat map was used to show the correlation between clustering and clinical parameters, and this was analyzed using a chi-square test. Significance analysis was performed using a two-sided unpaired t test. Results:A total of 35 up-regulated genes and 19 down-regulated genes were identified. These genes were mainly involved in biological processes and signaling pathways related to ferroptosis, oxidative stress and fatty acid metabolism. A total of 14 ferroptosis-related DEGs were identified to be associated with prognosis. The clusterring effect was best when hepatocellular carcinoma patients were divided into two subgroups. The survival rate of cluster 2 was lower than that of cluster 1 ( P<0.05). There was no significant difference in the tumor immune dysfunction and exclusion (TIDE) score between cluster 2 and cluster 1 ( P=0.43). Cluster 1 exhibited higher levels of immune cell infiltration, particularly CD4 + T cells ( P<0.01). The expression levels of 10 major histocompatibility complex (MHC) molecule-related genes were higher in cluster 1. The angiogenesis activity score ( P=0.048) and stemness score ( P=0.038) of cluster 2 were increased, and the expression levels of programmed death-1 ( PDCD1) and cytotoxic T lymphocyte-associated antigen-4 ( CTLA-4) in cluster 2 (5.924±0.013 and 5.475±0.042) were higher than those in cluster 1 (4.539±0.143 and 4.372±0.176) (both P<0.05). The expression levels of AURKA, glucose-6-phosphate dehydrogenease ( G6PD), ACACA, GABA type A receptor associated protein like 1 ( GABARAPL1) and ARRDC3 were correlated with the T stage, clinical stage and survival status of hepatocellular carcinoma. The survival rate of the high-risk group was lower than that of the low-risk group with time ( P<0.01). The area under the curve of the risk characteristics at 1, 3 and 5 years was 0.797, 0.717 and 0.639, respectively. The actual survival time 1, 3, and 5 years was highly consistent with the corresponding predicted survival time. The levels of memory B cell infiltration, angiogenesis activity score and cell stemness score, programmed death-ligand 1, CTLA-4, hepatitis A virus cell receptor 2, lymphocyte activation gene 3 and PDCD1 gene expression (0.013 8±0.036 0, 0.884±0.212, 0.387±0.135, 6.273±0.228, 5.847±0.331, 8.179±0.259, 6.859±0.263 and 5.142±0.326) in the high-risk group were higher than those in the low-risk group (0.001 5±0.021 0, 0.874±0.132, 0.298±0.125, 5.866±0.132, 3.742±0.237, 7.236±0.321, 6.324±0.242 and 4.513±0.211) ( P<0.05, 0.01). The expression levels of MHC molecule-related genes in the high-risk group were also higher than those in the low-risk group ( P<0.05, 0.01), while the infiltration levels of resting mast cells, activated natural killer cells, and resting natural killer cells (0.043 2±0.135 0, 0.032 1±0.143 0 and 0.016 3±0.001 9) and the TIDE score (0.072 0±0.018 0) in the high-risk group were lower than those in the low-risk group (0.054 9±0.023 0, 0.042 7±0.017 0, 0.024 6±0.021 2 and 0.094 0±0.013 5) ( P<0.05, 0.01). The top five genes with the highest mutation frequency in the high-risk group were tumor protein P53 ( TP53, 43%), titin ( TTN, 21%), catenin beta 1 ( CTNNB1, 20%), mucin 16 ( MUC16, 18%) and piccolo presynaptic cytomatrix protein ( PCLO, 11%). The top five genes with the highest mutation frequency in the low-risk group were CTNNB1 (30%), TTN (24%), albumin ( ALB, 16%), MUC16 (15%) and PCLO (11%). The cube protein and PCLO showed the co-occurrence of gene mutations in the high-risk group, while MUC16 and axis 1 protein showed the co-occurrence of gene mutations in the low-risk group. There was no significant difference in tumor mutation burden (TMB) between the high-risk group (1.374±0.026) and the low-risk group (1.303±0.081) ( P=0.073). There was no significant difference in survival time between the high-TMB group (2.3 years) and the low-TMB group (3.8 years) ( P=0.293). The mutation rates of AURKA, G6PD, ACACA, GABARAPL1 and ARRDC3 genes (2.0%, 2.0%, 4.0%, 0.3% and 0.6%) were relatively low. The relative expression levels of Aurka, Acaca and Arrdc3 mRNA in Hepa1-6 cells (13.331±0.000, 6.619±0.000 and 1.209±0.002) were higher than those in mouse primary hepatocytes (1.000±0.000, 1.000±0.000 and 1.000±0.000) (all P<0.01). The relative expression levels of AURKA, ACACA and ARRDC3 mRNA in tumor tissues of patients with hepatocellular carcinoma (2.102±0.365, 2.476±0.351 and 11.460±9.189) were higher than those in adjacent normal tissues of patients with hepatocellular carcinoma (1.122±0.648, 0.831±0.935 and 0.852±0.171) ( P<0.05, 0.01). Conclusions:This study constructed a prognostic signature comprising five ferroptosis-related genes ( AURKA, G6PD, ACACA, GABARAPL1, and ARRDC3) that is highly correlated with clinical hepatocellular carcinoma data. This study highlights the significance of ferroptosis-related genes as prognostic markers for hepatocellular carcinoma and provides insights into the complex relationship between hepatocellular carcinoma, ferroptosis, and the immune microenvironment.

Objective:To investigate the role of umbilical cord mesenchymal stem cell (MSC) -derived nanovesicles in hair regeneration.Methods:(1) Nanovesicles were prepared by continuously extruding umbilical cord MSCs through polycarbonate membranes, and were identified using transmission electron microscopy and nanoparticle tracking analysis. (2) Six C57BL/6 female mice with full-thickness skin wounds were randomly divided into a nanovesicle group (subcutaneously injected with nanovesicles once at the wound margin) and a control group (subcutaneously injected with an equal volume of phosphate-buffered saline [PBS] at the wound margin) ; skin samples were collected on day 16 for hematoxylin-eosin (HE) staining to assess wound healing and hair follicle regeneration. (3) Human hair follicle dermal papilla cells (DPCs) were isolated using a two-step enzyme method; the uptake of PKH26-pre-labeled nanovesicles by DPCs was observed by fluorescence microscopy; the proliferative activity of DPCs co-cultured with nanovesicles was evaluated using cell counting kit-8 (CCK8) and 5-ethynyl-2'-deoxyuridine (EdU) assays. (4) Six healthy C57BL/6 female mice were randomly divided into two groups after anesthesia, and subcutaneously injected with either fluorescent dye DIR-pre-labeled nanovesicles or PBS; an in vivo imaging system was used to observe the uptake and metabolism of nanovesicles in the mouse skin. (5) Twenty-four C57BL/6 female mice with depilated backs were randomly divided into a nanovesicle group (subcutaneously injected with nanovesicles on days 0, 8, and 15) and a control group (subcutaneously injected with an equal volume of PBS at the same time points) ; skin samples were collected on days 4, 18, and 21 for HE staining to analyze differences in hair follicle cycling; transcriptome sequencing was performed on skin samples collected on day 4. Statistical analyses were conducted using the t test. Results:(1) Transmission electron microscopy showed that nanovesicles exhibited a spherical membranous structure with diameters of 141.3 ± 60.0 nm. (2) In 6 C57BL/6 female mice with full-thickness skin wounds, the wound area on day 12 was significantly smaller in the nanovesicle group (1.27 ± 0.50 mm 2) than in the control group (4.13 ± 1.03 mm 2, t = 4.34, P = 0.012). (3) Fluorescence microscopy revealed that nanovesicles were taken up by DPCs within 20 hours; the absorbance of DPCs was significantly higher in the nanovesicle group than in the control group ( t = 20.23, P < 0.001), and the percentage of EdU-positive cells was also significantly higher in the nanovesicle group (49.62% ± 6.45%) than in the control group (37.58% ± 3.42%, t = 3.69, P = 0.006). (4) In vivo imaging of the 6 C57BL/6 female mice showed strong fluorescence in the back of mice in the nanovesicle group on day 0, which markedly decreased by day 8, while no fluorescence was observed in the control group throughout the experiment. (5) Hair follicle cycle experiments on the 24 C57BL/6 female mice with depilated backs showed that the hair follicle length on day 4 after depilation was significantly longer in the nanovesicle group (368.00 ± 63.17 μm) than in the control group (266.90 ± 34.41 μm, t = 9.87, P < 0.001), and the hair bulb diameter was also significantly longer in the nanovesicle group (54.83 ± 10.32 μm) than in the control group (39.12 ± 7.54 μm, t = 16.02, P < 0.001) ; on day 18, the nanovesicle group showed a significantly higher hair follicle density (19.12 ± 0.90) compared with the control group (11.07 ± 1.51, t = 7.92, P = 0.001) ; on day 21, 46.13% ± 8.64% of hair follicles in the nanovesicle group remained in the anagen phase Ⅵ to the catagen phase Ⅱ, and 46.24% ± 3.29% were in the catagen phases Ⅲ to Ⅳ, while 78.89% ± 18.36% of hair follicles in the control group were in the telogen phases Ⅶ to Ⅷ. Transcriptome sequencing showed that differentially expressed genes in the nanovesicle group were significantly positively enriched in the keratinization process (NES = 2.23, P < 0.001) . Conclusion:Umbilical cord MSC-derived nanovesicles could promote the proliferation of DPCs, advance the entry of hair follicles into the anagen phase, delay their entry into the catagen phase, and induce hair regeneration.

Objective:To investigate the role of umbilical cord mesenchymal stem cell (MSC) -derived nanovesicles in hair regeneration.Methods:(1) Nanovesicles were prepared by continuously extruding umbilical cord MSCs through polycarbonate membranes, and were identified using transmission electron microscopy and nanoparticle tracking analysis. (2) Six C57BL/6 female mice with full-thickness skin wounds were randomly divided into a nanovesicle group (subcutaneously injected with nanovesicles once at the wound margin) and a control group (subcutaneously injected with an equal volume of phosphate-buffered saline [PBS] at the wound margin) ; skin samples were collected on day 16 for hematoxylin-eosin (HE) staining to assess wound healing and hair follicle regeneration. (3) Human hair follicle dermal papilla cells (DPCs) were isolated using a two-step enzyme method; the uptake of PKH26-pre-labeled nanovesicles by DPCs was observed by fluorescence microscopy; the proliferative activity of DPCs co-cultured with nanovesicles was evaluated using cell counting kit-8 (CCK8) and 5-ethynyl-2'-deoxyuridine (EdU) assays. (4) Six healthy C57BL/6 female mice were randomly divided into two groups after anesthesia, and subcutaneously injected with either fluorescent dye DIR-pre-labeled nanovesicles or PBS; an in vivo imaging system was used to observe the uptake and metabolism of nanovesicles in the mouse skin. (5) Twenty-four C57BL/6 female mice with depilated backs were randomly divided into a nanovesicle group (subcutaneously injected with nanovesicles on days 0, 8, and 15) and a control group (subcutaneously injected with an equal volume of PBS at the same time points) ; skin samples were collected on days 4, 18, and 21 for HE staining to analyze differences in hair follicle cycling; transcriptome sequencing was performed on skin samples collected on day 4. Statistical analyses were conducted using the t test. Results:(1) Transmission electron microscopy showed that nanovesicles exhibited a spherical membranous structure with diameters of 141.3 ± 60.0 nm. (2) In 6 C57BL/6 female mice with full-thickness skin wounds, the wound area on day 12 was significantly smaller in the nanovesicle group (1.27 ± 0.50 mm 2) than in the control group (4.13 ± 1.03 mm 2, t = 4.34, P = 0.012). (3) Fluorescence microscopy revealed that nanovesicles were taken up by DPCs within 20 hours; the absorbance of DPCs was significantly higher in the nanovesicle group than in the control group ( t = 20.23, P < 0.001), and the percentage of EdU-positive cells was also significantly higher in the nanovesicle group (49.62% ± 6.45%) than in the control group (37.58% ± 3.42%, t = 3.69, P = 0.006). (4) In vivo imaging of the 6 C57BL/6 female mice showed strong fluorescence in the back of mice in the nanovesicle group on day 0, which markedly decreased by day 8, while no fluorescence was observed in the control group throughout the experiment. (5) Hair follicle cycle experiments on the 24 C57BL/6 female mice with depilated backs showed that the hair follicle length on day 4 after depilation was significantly longer in the nanovesicle group (368.00 ± 63.17 μm) than in the control group (266.90 ± 34.41 μm, t = 9.87, P < 0.001), and the hair bulb diameter was also significantly longer in the nanovesicle group (54.83 ± 10.32 μm) than in the control group (39.12 ± 7.54 μm, t = 16.02, P < 0.001) ; on day 18, the nanovesicle group showed a significantly higher hair follicle density (19.12 ± 0.90) compared with the control group (11.07 ± 1.51, t = 7.92, P = 0.001) ; on day 21, 46.13% ± 8.64% of hair follicles in the nanovesicle group remained in the anagen phase Ⅵ to the catagen phase Ⅱ, and 46.24% ± 3.29% were in the catagen phases Ⅲ to Ⅳ, while 78.89% ± 18.36% of hair follicles in the control group were in the telogen phases Ⅶ to Ⅷ. Transcriptome sequencing showed that differentially expressed genes in the nanovesicle group were significantly positively enriched in the keratinization process (NES = 2.23, P < 0.001) . Conclusion:Umbilical cord MSC-derived nanovesicles could promote the proliferation of DPCs, advance the entry of hair follicles into the anagen phase, delay their entry into the catagen phase, and induce hair regeneration.

[Objective]To explore the medication patterns of WU's tumor school for anti-metastasis through"enhancing health and promoting detoxification"based on data mining.[Methods]Systematically review the literature related to the prescription and medication patterns of WU Liangcun in the treatment of tumor metastasis,and summarize his clinical experience in the use of the"enhancing health and promoting detoxification"method.Complete case records of lung cancer,gastric cancer and colorectal cancer metastasis treated by SHEN Minhe,WANG Binbin and RUAN Shanming from January 1,2020 to August 31,2023 were collected.Data mining analysis was performed on the effective prescriptions by using the Traditional Chinese Medicine Inheritance Computer System(V3.5).[Results]A total of 3 023 prescriptions from SHEN Minhe were included,involving 344 types of Chinese herbs.Codonopsis Radix was frequently used,and common herb pairs included Codonopsis Radix,Poria cocos-Atractylodis Macrocephalae Rhizoma.WANG Binbin's 2 654 prescriptions involved 370 types of Chinese herbs.Paeoniae Radix Alba was frequently used,and common pairs included Coicis Semen,Fagopyri Dibotryis Rhizoma-Hedyotis diffusa Willd.RUAN Shanming's 1 182 prescriptions included 257 types of herbs.Astragali Radix was frequently used,and common pairs included Astragali Radix,Alismatis Rhizoma-Scutellariae Radix.WU's tumor school prefers to use herbs with sweet,bitter and acrid flavors,as well as neutral,cold and warm properties,avoiding excessive purgation or tonification and extreme cold or heat.They advocated for gentle tonification and gradual adjustment,as well as a balanced combination of cold and heat.In terms of meridian tropism,they focused on using herbs that entered the spleen and lung meridians,targeting the pathogenesis of spleen and stomach deficiency,which led to the accumulation of dampness and phlegm,emphasized the simultaneous regulation of the spleen and lung,to strengthen the spleen and stomach,eliminate dampness and phlegm,regulate the flow of Qi and clear cancerous toxins.[Conclusion]WU's tumor school has been passed down for three generations,following the principle of"enhancing health and promoting detoxification".The overall medication is balanced and tailored to different types of malignant tumors,demonstrating significant clinical efficacy.

[Objective]To explore the medication patterns of WU's tumor school for anti-metastasis through"enhancing health and promoting detoxification"based on data mining.[Methods]Systematically review the literature related to the prescription and medication patterns of WU Liangcun in the treatment of tumor metastasis,and summarize his clinical experience in the use of the"enhancing health and promoting detoxification"method.Complete case records of lung cancer,gastric cancer and colorectal cancer metastasis treated by SHEN Minhe,WANG Binbin and RUAN Shanming from January 1,2020 to August 31,2023 were collected.Data mining analysis was performed on the effective prescriptions by using the Traditional Chinese Medicine Inheritance Computer System(V3.5).[Results]A total of 3 023 prescriptions from SHEN Minhe were included,involving 344 types of Chinese herbs.Codonopsis Radix was frequently used,and common herb pairs included Codonopsis Radix,Poria cocos-Atractylodis Macrocephalae Rhizoma.WANG Binbin's 2 654 prescriptions involved 370 types of Chinese herbs.Paeoniae Radix Alba was frequently used,and common pairs included Coicis Semen,Fagopyri Dibotryis Rhizoma-Hedyotis diffusa Willd.RUAN Shanming's 1 182 prescriptions included 257 types of herbs.Astragali Radix was frequently used,and common pairs included Astragali Radix,Alismatis Rhizoma-Scutellariae Radix.WU's tumor school prefers to use herbs with sweet,bitter and acrid flavors,as well as neutral,cold and warm properties,avoiding excessive purgation or tonification and extreme cold or heat.They advocated for gentle tonification and gradual adjustment,as well as a balanced combination of cold and heat.In terms of meridian tropism,they focused on using herbs that entered the spleen and lung meridians,targeting the pathogenesis of spleen and stomach deficiency,which led to the accumulation of dampness and phlegm,emphasized the simultaneous regulation of the spleen and lung,to strengthen the spleen and stomach,eliminate dampness and phlegm,regulate the flow of Qi and clear cancerous toxins.[Conclusion]WU's tumor school has been passed down for three generations,following the principle of"enhancing health and promoting detoxification".The overall medication is balanced and tailored to different types of malignant tumors,demonstrating significant clinical efficacy.

OBJECTIVE To investigate the factors influencing the changes in purchasing quantity in the procurement varieties of the first batch of volume-based drug centralized procurement （hereinafter referred to as centralized procurement）. METHODS Using 25 procurement varieties of the “4+7” policy as research objects， the changes in purchasing quantity of procurement varieties were analyzed before and after the implementation of the “4+7” pilot， renewal and expansion policies. The influential factors were determined from the three levels of drugs， medical institutions and regions； and the multiple linear regression model was used to analyze the influential factors for the changes in the purchasing quantity of procurement varieties. RESULTS Before and after the implementation of the “4+7” pilot， renewal and expansion policies， the purchasing quantity increased by 52.1， －0.2， 85.8 ten thousand DDDs on average， compared with base period. During pilot， renewal and expansion period， DDDc decrease in procurement varieties was positively correlated with the increase in purchasing quantity （P＜0.01）. During the pilot and renewal period， the number of absolutely alternative varieties was positively correlated with the increase in purchasing quantity （P＜0.1）. During the pilot and expansion period， the number of alternative varieties to a certain extent was negatively correlated with the increase in purchasing quantity （P＜0.05）. During the renewal period， the increment of purchasing quantity in tertiary hospitals was smaller than that of primary medical institutions （P＜0.05）. CONCLUSIONS There is a relationship between the decline of DDDc and the changes in the purchasing quantity， that is， the more the drug price dropped， the more the purchasing quantity increased. The number of alternative varieties for centralized procurement will affect the changes in their purchasing quantity， but it is not always stable. With the implementation of the policy， the volume of primary medical institutions gradually exceeds that of tertiary institutions， indicating that the consumption of centralized purchased varieties is transferred to the primary medical institutions， and centralized procurement has promoted the implementation of the hierarchical diagnosis and treatment system.

Background In the Global Burden of Disease research, it has been found that atmospheric fine particulate matter (PM_2.5) pollution significantly harms human health. Currently, there is limited research on polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) that exhibit high toxicity effects in PM_2.5 . Objective By studying the spatiotemporal distribution and variation characteristics of PCDD/Fs in PM_2.5 in Pudong area of Shanghai, to assess the associated population health risk. Methods This study set up 28 sampling points in Pudong area. One sample of PM_2.5 was collected during winter (February 2022) and summer (July 2022) at each site, with a sampling period lasting 24 h. The concentration of PM_2.5 was measured by membrane filter method, and the content of 17 kinds of 2,3,7,8-substituted chlorinated PCDD/Fs in the samples was analyzed using isotope dilution. Seasonal variations (winter and summer) in the concentrations of PM_2.5 and PCDD/Fs were evaluated, sources of PCDD/Fs pollution were tracing by principal component analysis, and health risks to the population from respiratory exposure to PCDD/Fs were estimated by VLIER-HUMAAN model. Results The PM_2.5 concentrations in the 28 samples ranged from 10 to 126 μg·m⁻³, while the concentrations of PCDD/Fs in PM_2.5 ranged from 58 to 2625 fg·m⁻³. The concentration of PM_2.5 during winter (11-126 μg·m⁻³) was higher than that during summer (10-60 μg·m⁻³). The concentration range of PCDD/Fs in winter was from 58 to 2625 fg·m⁻³, which corresponded to a range of toxic equivalent quantity (WHO-TEQ) concentration from 2.99 to 40.97 fg·m⁻³ when taking World Health Organization's toxic equivalency factor (WHO-TEQ); the concentration range of PCDD/Fs in summer was from 72 to 446 fg·m⁻³, which corresponded to a range of WHO-TEQ concentration from 2.66 to 16.61 fg·m⁻³. This range in summer was significantly lower than that observed in winter. The results of principal component analysis revealed that waste incineration was the primary source of PCDD/Fs in winter PM_2.5 in the area, whereas traffic emissions emerged as the main source in summer. The assessment of Pudong residents' respiratory exposure to PCDD/Fs in PM_2.5 showed significantly higher exposure of children in summer and winter than that of adults, indicating higher susceptibility of children to air pollutants. Both the hazard ratios (HR) for children and adults were below 1, while the cancer risks (CR) ranged from 8.41×10⁻⁸ to 2.35×10⁻⁶. Notably, during winter, the CR at 4 locations slightly exceeds 1×10⁻⁶, indicating a potential carcinogenic risk. Conclusion The overall pollution level of PCDD/Fs in PM_2.5 in Pudong area is relatively low, but it shows clear seasonal patterns. Waste incineration and traffic are the main sources of PCDD/Fs in PM_2.5 in the area. Although the cancer risk of exposure to PCDD/Fs in PM_2.5 for children or adults is relatively low, there is a certain risk at some locations in winter, necessitating additional monitoring and control.

Objective:Based on the specific cleavage and non-specific "trans-cleavage" activities of the clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein(CRISPR/Cas13), we established a visually amplification-free rapid detection technique of 2019-nCoV nucleic acid. This technique is easily processed with a low detection limit and good specificity.Methods:According to the 2019-nCoV gene sequence, specific CRISPR RNAs were screened and designed by bioinformatics analysis, and then synthesized as universal signal-strained RNA transcription targets in vitro to establish and optimize the reaction system. Moreover, the 2019-nCoV pseudoviral nucleic acid was used as a standard substance to evaluate the detection limit. A total of 65 positive samples were collected from various 2019-nCoV variants, while 48 negative samples included other clinically common respiratory pathogens, such as influenza A virus, influenza B virus, human parainfluenza virus, Klebsiella pneumonia, etc. All samples were tested by quantitative PCR (qPCR), digital PCR, and the method established in this study. The sensitivity and specificity of the newly established method were analyzed and evaluated. Results:With the newly established technique, the detection time for 2019-nCoV nucleic acid could be minimized to 6 minutes. In addition, the detection limit was 14 copies/μl when assisted by the displaying instrument, whereas it increased to 28 copies/μl with the naked eye. This technique had a sensitivity and specificity of 98.5% (66/67) and 100% (46/46) respectively, showing no statistically significant difference compared to the gold standard qPCR( P=1). Conclusions:This study has successfully established a CRISPR/Cas13a-based visually rapid detection technique for 2019-nCoV nucleic acid. This technique offers the advantages of a simple process, convenient operation, low environmental operating requirements, a detection limit close to qPCR, and a strong potential for on-site testing applications.

Objective:To describe the clinical characteristics of patients with autoimmune gastritis (AIG), and to further explore the clinical differences between AIG patients with positive Helicobacter pylori ( H. pylori) and with negative H. pylori, and to reveal the significance of H. pylori in AIG patients. Methods:From January 1, 2020 to December 31, 2023, 112 patients visited Taizhou Hospital of Zhejiang Province who underwent endoscopy examinations and AIG-related antibody tests were retrospectively enrolled. Among them, 34 cases were complicated with H. pylori infection ( H. pylori-positive group) and 78 cases were not complicated with H. pylori infection ( H. pylori-negative group). Anemia status, the positive rates of AIG antibodies including anti-parietal cell antibody (PCA) and intrinsic factor antibody (IFA), gastric function markers such as gastrin-17, pepsinogen Ⅰ (PGⅠ) and the ratio of PGⅠ to PG Ⅱ, vitamin B12 and folic acid levels, as well as the manifestations under gastroscopy, thyroid function indicators, and results of thyroid ultrasound examination were comparatively analyzed between H. pylori-positive group and H. pylori-negative group. Independent samples t-test, Mann-Whitney U test, and chi-square test were used for statistical analysis. Results:Among the 112 patients with AIG, 30 cases were males and 82 cases were females, with the age of (59.3±10.1) years old. Twenty-three (20.5%) AIG patients were complicated with iron deficiency anemia and 13 (11.6%) AIG patients were complicated with megaloblastic anemia. The proportion of patients complicated with megaloblastic anemia of H. pylori-positive group was higher than that of H. pylori-negative group (23.5%, 8/34 vs. 6.4%, 5/78), and the difference was statistically significant ( χ2=5.20, P=0.023). The positive rates of PCA, IFA, thyroid peroxidase antibody, and thyroglobulin antibody were 98.2% (110/112), 27.5% (28/102), 75.0% (24/32), and 62.5% (20/32), respectively. The gastrin-17 level of 94.2% (97/103) AIG patients was more than 5 times the normal upper limit; and the vitamin B12 level of 24.4% (22/90) AIG patients decreased. There were 84.0% (42/50) of AIG patients complicated with thyroid nodules or echo changes under ultrasound, and 18.8% (12/64) of AIG patients had thyroid function changes. In addition to reverse atrophy under endoscopy, yellow-white turbid mucus was found in 51.8% (58/112) AIG patients, 51 cases (45.5%) combined with proliferative polyps, 8 cases (7.1%) combined with gastric neuroendocrine tumors and 7 cases (6.2%) combined with gastric adenoma or adenocarcinoma. The proportion of patients with adenoma or adenocarcinoma in H. pylori-positive group was higher than that of H. pylori-negative group (14.7%, 5/34 vs. 2.6%, 2/78), and the difference was statistically significant ( χ2=4.07, P=0.044). Conclusions:When unexplained anemia occurs clinically, inverse atrophy or gastric neuroendocrine tumors presented under endoscopy, positive gastric autoantibodies detected, the diagnosis of AIG should be considered. The eradication of H. pylori still remains as the key to the treatment of AIG patients.

Objective:To describe the clinical characteristics of patients with autoimmune gastritis (AIG), and to further explore the clinical differences between AIG patients with positive Helicobacter pylori ( H. pylori) and with negative H. pylori, and to reveal the significance of H. pylori in AIG patients. Methods:From January 1, 2020 to December 31, 2023, 112 patients visited Taizhou Hospital of Zhejiang Province who underwent endoscopy examinations and AIG-related antibody tests were retrospectively enrolled. Among them, 34 cases were complicated with H. pylori infection ( H. pylori-positive group) and 78 cases were not complicated with H. pylori infection ( H. pylori-negative group). Anemia status, the positive rates of AIG antibodies including anti-parietal cell antibody (PCA) and intrinsic factor antibody (IFA), gastric function markers such as gastrin-17, pepsinogen Ⅰ (PGⅠ) and the ratio of PGⅠ to PG Ⅱ, vitamin B12 and folic acid levels, as well as the manifestations under gastroscopy, thyroid function indicators, and results of thyroid ultrasound examination were comparatively analyzed between H. pylori-positive group and H. pylori-negative group. Independent samples t-test, Mann-Whitney U test, and chi-square test were used for statistical analysis. Results:Among the 112 patients with AIG, 30 cases were males and 82 cases were females, with the age of (59.3±10.1) years old. Twenty-three (20.5%) AIG patients were complicated with iron deficiency anemia and 13 (11.6%) AIG patients were complicated with megaloblastic anemia. The proportion of patients complicated with megaloblastic anemia of H. pylori-positive group was higher than that of H. pylori-negative group (23.5%, 8/34 vs. 6.4%, 5/78), and the difference was statistically significant ( χ2=5.20, P=0.023). The positive rates of PCA, IFA, thyroid peroxidase antibody, and thyroglobulin antibody were 98.2% (110/112), 27.5% (28/102), 75.0% (24/32), and 62.5% (20/32), respectively. The gastrin-17 level of 94.2% (97/103) AIG patients was more than 5 times the normal upper limit; and the vitamin B12 level of 24.4% (22/90) AIG patients decreased. There were 84.0% (42/50) of AIG patients complicated with thyroid nodules or echo changes under ultrasound, and 18.8% (12/64) of AIG patients had thyroid function changes. In addition to reverse atrophy under endoscopy, yellow-white turbid mucus was found in 51.8% (58/112) AIG patients, 51 cases (45.5%) combined with proliferative polyps, 8 cases (7.1%) combined with gastric neuroendocrine tumors and 7 cases (6.2%) combined with gastric adenoma or adenocarcinoma. The proportion of patients with adenoma or adenocarcinoma in H. pylori-positive group was higher than that of H. pylori-negative group (14.7%, 5/34 vs. 2.6%, 2/78), and the difference was statistically significant ( χ2=4.07, P=0.044). Conclusions:When unexplained anemia occurs clinically, inverse atrophy or gastric neuroendocrine tumors presented under endoscopy, positive gastric autoantibodies detected, the diagnosis of AIG should be considered. The eradication of H. pylori still remains as the key to the treatment of AIG patients.