ObjectiveTo investigate the mechanism of Qinggan Jianpi Houxue prescription against hepatic fibrosis in mice using metabolomics. MethodsA hepatic fibrosis mouse model was established by intraperitoneal injection of 20% carbon tetrachloride （CCl₄， diluted with olive oil at a ratio of 4∶1） at a dose of 2.5 mL·kg^-1 for 4 weeks. A total of 54 C57BL/6J mice were randomly divided into the blank control group， model group， Qinggan Jianpi Houxue prescription high-dose group （47.32 g·kg^-1）， medium-dose group （23.66 g·kg^-1）， low-dose group （11.83 g·kg^-1）， and Biejiajian pill group （1.365 g·kg^-1）， with 9 mice in each group. Concurrently with modeling， each administration group received the corresponding dose continuously for 4 weeks， once daily. After 4 weeks， liver tissues and blood samples were collected for liver function and hepatic pathological examinations. Non-targeted metabolomics was employed to detect hepatic metabolites. Principal component analysis （PCA）， partial least squares discriminant analysis （PLS-DA） and orthogonal partial least squares discriminant analysis （OPLS-DA） were performed to explore the metabolites and their involved metabolic pathways in the liver tissues of mice in the blank control group， model group and Qinggan Jianpi Houxue prescription high-dose group. Western blot validation was conducted at both the cellular and animal levels. ResultsCompared with the blank control group， the model group showed significantly increased levels of alanine aminotransferase （ALT） and aspartate aminotransferase （AST） （P<0.01）， along with increased inflammatory cell infiltration， and formation of fibrous septa. The predominantly enriched pathway was the forkhead box O （FoxO） signaling pathway. Compared with the model group， the Qinggan Jianpi Huoxue prescription high-dose group exhibited significantly decreased ALT and AST levels （P<0.01）. In all administration groups of Qinggan Jianpi Huoxue prescription， inflammatory cell infiltration was reduced， fibrous septa were narrowed and diminished， collagen fibers were reduced， and the collagen area was significantly decreased. Non-targeted metabolomics identified 79 differential metabolites， among which 49 were up-regulated and 30 down-regulated， primarily enriched in the tricarboxylic acid （TCA） cycle and pyruvate metabolism pathways. Western blot results demonstrated that， compared with the blank control group， the model group exhibited significantly upregulated protein expressions of α-smooth muscle actin （α-SMA）， lactate dehydrogenase A （LDHA）， FoxO1， pyruvate kinase M2 （PKM2）， hexokinase 2 （HK2）， and phosphofructokinase 1 （PFK1） in liver tissues （P<0.05， P<0.01）. In hepatic stellate cells， the glucose concentration increased， and the protein expression of α-SMA， LDHA， FoxO1， PKM2， glucose transporter 1 （GLUT1）， and PFK1 were significantly up-regulated （P<0.05， P<0.01）. Conversely， compared with the model group， the Qinggan Jianpi Huoxue prescription high-dose group showed significantly downregulated protein expression of α-SMA， LDHA， FoxO1， PKM2， HK2， and PFK1 in liver tissues （P<0.05， P<0.01）. In hepatic stellate cells， the glucose concentration decreased， and the protein expression of α-SMA， LDHA， FoxO1， PKM2， GLUT1， and PFK1 were significantly down-regulated （P<0.05， P<0.01）. ConclusionQinggan Jianpi Huoxue prescription can alleviate the pathological progression of hepatic fibrosis in mice， potentially by regulating the TCA cycle and pyruvate metabolism through FoxO1， and by modulating glycolytic metabolic reprogramming in hepatic stellate cells， thereby promoting liver tissue repair.

Tuberculosis is a infectious disease caused by Mycobacterium tuberculosis( Mtb), which seriously threatens human health. Innate immune cells can synergistically resist the invasion of Mtb by phagocytosing pathogens, releasing inflammatory cytokines, and activating other immune cells, playing a crucial role in the host′s defense against Mtb infection. A comprehensive and in-depth understanding of the role of innate immune cells in Mtb infection will help reveal the complexity of immune response and understand the mechanism of early elimination of Mtb and immune escape of Mtb, and provide new ideas and strategies for the prevention and treatment of tuberculosis. This article will provide an overview of the research progress on the mechanisms of several important innate immune cells, including macrophages, neutrophils, natural killer cells, dendritic cells, and constant natural killer T cells, in Mtb infection.

Objective:To analyze the incidence and spatiotemporal clustering of hepatitis B in China from 2006 to 2020 and provide reference for hepatitis B prevention and control.Methods:The incidence data of hepatitis B in 31 provinces (autonomous regions and municipalities) from 2006 to 2020 were collected from National Notifiable Infectious Disease Reporting System of China Information System for Disease Control and Prevention. The incidence trend analysis was conducted by using software Joinpoint 5.0.2, and the spatiotemporal scan analysis was performed by using software SaTScan 10.1.2.Results:From 2006 to 2020, a total of 1 049 546 cases of acute hepatitis B were reported in China. The average annual reported incidence rate was 5.17/100 000. The reported incidence rate showed a decreasing trend during this period. The incidence decreased from 3.00/100 000 to 0.41/100 000 in age group 0-14 years, from 14.15/100 000 to 3.44/100 000 in age group 15-34 years, and from 6.87/100 000 to 3.72/100 000 in age group ≥35 years, the differences were all significant (all P<0.001). From 2006 to 2020, a total of 10 732 017 cases of chronic hepatitis B were reported in China. The average annual reported incidence rate was 52.85/100 000. The reported incidence of chronic hepatitis B varied in different age groups, which decreased from 11.38/100 000 to 2.18/100 000 in age group 0-14 years, and from 73.17/100 000 to 61.40/100 000 in age group 15-34 years, while increased from 48.07/100 000 to 90.75/100 000 in age group ≥35 years, the differences were all significant (all P<0.05). Spatiotemporal scan analysis indicated that the age of reported acute hepatitis B cases became older over time, and the regions with high-incidence gradually shifted from western China to southwestern China. The overall reported incidence of chronic hepatitis B in those aged ≥35 years showed an upward trend, and the regions with high-incidence were mainly found in coastal area in southeastern China and in southwestern China. Conclusions:From 2006 to 2020, the overall reported incidence of acute hepatitis B in China showed a continuous downward trend, while the reported incidence of chronic hepatitis B in those aged ≥35 years showed an upward trend. It indicated that the need to improve the diagnosis and treatment of chronic hepatitis B in adults in coastal area in southeastern China and southwestern China.

Objective:To evaluate the role of docosahexaenoic acid (DHA) in long-term cognitive impairment after multiple sevoflurane anesthesia in newborn mice.Methods:Clean-grade healthy male C57BL/6 mice, aged 6 days, were used in this study. Ten mice were divided into 2 groups ( n=5 each) by the random number table method: control group (group C) and sevoflurane group (group S). The animals inhaled 3% sevoflurane for 2 h at 6, 7 and 8 days after birth. The DHA content was detected by ultra-high performance liquid chromatography-mass spectrometry at 9 days of age. Fifty-two mice were selected and divided into 4 groups ( n=13 each) by a random number table method: control+ normal saline group (group C+ S), sevoflurane anesthesia + normal saline group (group S+ S), control+ DHA group (group C+ D), and sevoflurane anesthesia+ DHA group (group S+ D). The sevoflurane anesthesia method was the same as the one mentioned above. DHA 50 mg/kg was administered by intragastric gavage from postnatal days 6-19 (at 6, 7 and 8 days after birth, 2 h before anesthesia) in C+ D and S+ D groups. The equal volume of normal saline was given instead in C+ S group and S+ S group. The novel object recognition test was conducted at 37 days of age, and the Morris water maze test was performed at 42 days of age. The corpus callosum and hippocampal tissues were isolated at 47 days of age for examination of the ultrastructure of myelin (with a transmission electron microscope) and for determination of the expression of myelin basic protein (MBP) in hippocampal tissues (by Western blot). The G-ratio was calculated. Results:Compared with group C, the content of DHA in hippocampal tissues was significantly decreased in group S ( P<0.05). Compared with group C+ S, the discrimination index was significantly decreased, the percentage of duration of staying at the target platform quadrant and the number of crossing the original platform were decreased, the expression of MBP was down-regulated, and the G-ratio in the original platform and hippocampus was increased in S+ S group ( P<0.05). Compared with group S+ S, the discrimination index was significantly increased, the percentage of duration of staying at the target platform quadrant and the number of crossing the original platform were increased, the expression of MBP was up-regulated, and the G-ratio in the original platform and hippocampus was decreased in S+ D group ( P<0.05). Conclusions:The mechanism of long-term cognitive impairment following multiple sevoflurane anesthesia may be related to a decrease in the content of DHA, which subsequently leads to myelin structural damage in neonatal mice.

Objective:To analyze the genetic characteristics and evolutionary origin of hepatitis B virus (HBV) subgenotype D3 in China in 2020.Methods:Serum samples and demographic details from patients infected with HBV D3 subgenotype were collected. HBV genomic sequences were obtained by nested PCR amplification and subsequent sequencing. Phylogenetic analysis, nucleotide homology, amino acid mutation and evolution rate of the S protein were conducted by comparing with reference sequence using bioinformatics tools.Results:The complete HBV gene sequences of 14 samples of D3 subtype HBV were obtained. Compared with 97 reference sequences, it was found that the sequences with the highest homology were from India, Mongolia, Iran and China, with the homology ranging from 96.0% to 97.9%. Mutations of 24 amino acids were found in 14 strains of D3 subtype. Among them, T131A, Y134F and T140I were associated with immune escape-related mutations. The genetic diversity of HBV D3 subtype increased slowly before 1975, remained relatively constant from 1975 to 2000, and began to decline after 2000. Evolutionary rate analysis showed that samples QGLD D3-02 and 03 originated from a common ancestor with the Iranian reference strain in 1872, and the other 12 samples QGLD D3-04-17 originated from a common ancestor with the Mongolian reference strain in 1843.Conclusions:The gene sequence of HBV D3 subtype in China had the highest homology with reference sequences from India, Iran, Mongolia and China. Evolutionary rate analysis revealed that 14 cases of HBV D3 subtype originated from a common ancestor with reference strains from Mongolia and Iran, which enriched the sequence and evolution information of HBV D3 subtype and provided a reference basis for the molecular epidemiological study of HBV.

Objective:To analyze the incidence and spatiotemporal clustering of hepatitis B in China from 2006 to 2020 and provide reference for hepatitis B prevention and control.Methods:The incidence data of hepatitis B in 31 provinces (autonomous regions and municipalities) from 2006 to 2020 were collected from National Notifiable Infectious Disease Reporting System of China Information System for Disease Control and Prevention. The incidence trend analysis was conducted by using software Joinpoint 5.0.2, and the spatiotemporal scan analysis was performed by using software SaTScan 10.1.2.Results:From 2006 to 2020, a total of 1 049 546 cases of acute hepatitis B were reported in China. The average annual reported incidence rate was 5.17/100 000. The reported incidence rate showed a decreasing trend during this period. The incidence decreased from 3.00/100 000 to 0.41/100 000 in age group 0-14 years, from 14.15/100 000 to 3.44/100 000 in age group 15-34 years, and from 6.87/100 000 to 3.72/100 000 in age group ≥35 years, the differences were all significant (all P<0.001). From 2006 to 2020, a total of 10 732 017 cases of chronic hepatitis B were reported in China. The average annual reported incidence rate was 52.85/100 000. The reported incidence of chronic hepatitis B varied in different age groups, which decreased from 11.38/100 000 to 2.18/100 000 in age group 0-14 years, and from 73.17/100 000 to 61.40/100 000 in age group 15-34 years, while increased from 48.07/100 000 to 90.75/100 000 in age group ≥35 years, the differences were all significant (all P<0.05). Spatiotemporal scan analysis indicated that the age of reported acute hepatitis B cases became older over time, and the regions with high-incidence gradually shifted from western China to southwestern China. The overall reported incidence of chronic hepatitis B in those aged ≥35 years showed an upward trend, and the regions with high-incidence were mainly found in coastal area in southeastern China and in southwestern China. Conclusions:From 2006 to 2020, the overall reported incidence of acute hepatitis B in China showed a continuous downward trend, while the reported incidence of chronic hepatitis B in those aged ≥35 years showed an upward trend. It indicated that the need to improve the diagnosis and treatment of chronic hepatitis B in adults in coastal area in southeastern China and southwestern China.

Objective:To evaluate the role of docosahexaenoic acid (DHA) in long-term cognitive impairment after multiple sevoflurane anesthesia in newborn mice.Methods:Clean-grade healthy male C57BL/6 mice, aged 6 days, were used in this study. Ten mice were divided into 2 groups ( n=5 each) by the random number table method: control group (group C) and sevoflurane group (group S). The animals inhaled 3% sevoflurane for 2 h at 6, 7 and 8 days after birth. The DHA content was detected by ultra-high performance liquid chromatography-mass spectrometry at 9 days of age. Fifty-two mice were selected and divided into 4 groups ( n=13 each) by a random number table method: control+ normal saline group (group C+ S), sevoflurane anesthesia + normal saline group (group S+ S), control+ DHA group (group C+ D), and sevoflurane anesthesia+ DHA group (group S+ D). The sevoflurane anesthesia method was the same as the one mentioned above. DHA 50 mg/kg was administered by intragastric gavage from postnatal days 6-19 (at 6, 7 and 8 days after birth, 2 h before anesthesia) in C+ D and S+ D groups. The equal volume of normal saline was given instead in C+ S group and S+ S group. The novel object recognition test was conducted at 37 days of age, and the Morris water maze test was performed at 42 days of age. The corpus callosum and hippocampal tissues were isolated at 47 days of age for examination of the ultrastructure of myelin (with a transmission electron microscope) and for determination of the expression of myelin basic protein (MBP) in hippocampal tissues (by Western blot). The G-ratio was calculated. Results:Compared with group C, the content of DHA in hippocampal tissues was significantly decreased in group S ( P<0.05). Compared with group C+ S, the discrimination index was significantly decreased, the percentage of duration of staying at the target platform quadrant and the number of crossing the original platform were decreased, the expression of MBP was down-regulated, and the G-ratio in the original platform and hippocampus was increased in S+ S group ( P<0.05). Compared with group S+ S, the discrimination index was significantly increased, the percentage of duration of staying at the target platform quadrant and the number of crossing the original platform were increased, the expression of MBP was up-regulated, and the G-ratio in the original platform and hippocampus was decreased in S+ D group ( P<0.05). Conclusions:The mechanism of long-term cognitive impairment following multiple sevoflurane anesthesia may be related to a decrease in the content of DHA, which subsequently leads to myelin structural damage in neonatal mice.

Tuberculosis is a infectious disease caused by Mycobacterium tuberculosis( Mtb), which seriously threatens human health. Innate immune cells can synergistically resist the invasion of Mtb by phagocytosing pathogens, releasing inflammatory cytokines, and activating other immune cells, playing a crucial role in the host′s defense against Mtb infection. A comprehensive and in-depth understanding of the role of innate immune cells in Mtb infection will help reveal the complexity of immune response and understand the mechanism of early elimination of Mtb and immune escape of Mtb, and provide new ideas and strategies for the prevention and treatment of tuberculosis. This article will provide an overview of the research progress on the mechanisms of several important innate immune cells, including macrophages, neutrophils, natural killer cells, dendritic cells, and constant natural killer T cells, in Mtb infection.

Objective:To analyze the genetic characteristics and evolutionary origin of hepatitis B virus (HBV) subgenotype D3 in China in 2020.Methods:Serum samples and demographic details from patients infected with HBV D3 subgenotype were collected. HBV genomic sequences were obtained by nested PCR amplification and subsequent sequencing. Phylogenetic analysis, nucleotide homology, amino acid mutation and evolution rate of the S protein were conducted by comparing with reference sequence using bioinformatics tools.Results:The complete HBV gene sequences of 14 samples of D3 subtype HBV were obtained. Compared with 97 reference sequences, it was found that the sequences with the highest homology were from India, Mongolia, Iran and China, with the homology ranging from 96.0% to 97.9%. Mutations of 24 amino acids were found in 14 strains of D3 subtype. Among them, T131A, Y134F and T140I were associated with immune escape-related mutations. The genetic diversity of HBV D3 subtype increased slowly before 1975, remained relatively constant from 1975 to 2000, and began to decline after 2000. Evolutionary rate analysis showed that samples QGLD D3-02 and 03 originated from a common ancestor with the Iranian reference strain in 1872, and the other 12 samples QGLD D3-04-17 originated from a common ancestor with the Mongolian reference strain in 1843.Conclusions:The gene sequence of HBV D3 subtype in China had the highest homology with reference sequences from India, Iran, Mongolia and China. Evolutionary rate analysis revealed that 14 cases of HBV D3 subtype originated from a common ancestor with reference strains from Mongolia and Iran, which enriched the sequence and evolution information of HBV D3 subtype and provided a reference basis for the molecular epidemiological study of HBV.

Objective:To analyze the whole genome sequence and key site mutations of hepatitis B virus (HBV) in patients with different stages of disease progression, and to understand the relationship between HBV genetic characteristics and disease progression.Methods:Serum samples and basic information of hepatitis B patients with asymptomatic HBV carrier, chronic hepatitis B patients, cirrhosis patients and primary hepatocellular carcinoma patients were collected. Nested PCR was used to amplify the samples to obtain HBV whole gene sequences. Phylogenetic trees were constructed to determine the genotype of the samples, and gene mutations of the samples were analyzed combined with reference sequences of each type.Results:A total of 256 samples were successfully amplified, including 68 asymptomatic HBV carrier patients, 118 CHB patients, 15 LC patients and 55 HCC patients, and five genotypes (B, C, D, I and C/D) were detected. The result of comparative analysis showed that the mutation rate of 56 nucleotide sites was significantly different among the four groups ( P<0.05). In addition to the discovery of C105T, A1762T/G1764A and G1899A and other previously reported key site mutations, the mutation rates of T53A, C1485T and C1628T in newly diagnosed HCC group were significantly higher than those in other groups, and the mutation rates of T2150G and T2151C in asymptomatic HBV infection group were significantly higher than those in other groups. A total of 26 sequences were deleted, mainly distributed in the pre-C and pre-S regions. The deletion mutation rate in the HCC group was significantly higher than that in the other groups. Conclusions:The data of this study indicate that some nucleotide substitution mutations and deletion mutations may be closely related to the occurrence and development of HBV-related diseases, and HCC patients are more likely to have gene mutations than non-HCC patients. These result provide a reference for understanding the relationship between viral mutation and the progression of HBV infection-related diseases.