Objective To investigate the mechanism of Kuijie Ankang Decoction in regulating immune,autophagy and intestinal flora in the treatment of ulcerative colitis(UC).Methods UC mouse model was established by free drinking with sodium dextran sulfate.The mice were randomly divided into blank control group,model group,Kuijie Ankang Decoction group,salazine sulfopyridine(SASP)group and 3-methyladenine(3-MA)group,with 12 mice in each group.Each drug group was given corresponding drugs for gavage,the blank control group and model group were given the same volume of distilled water for gavage for 7 days.The general condition of mice was observed and the disease activity index(DAI)was scored,the morphology of colon tissue was observed by HE staining,the contents of TNF-α,IL-1β,IL-6,IL-8 and IL-10 in colon tissue were detected by ELISA,the mRNA and protein expressions of LC3,Beclin-1 and p62 in colon tissue were detected by RT-qPCR and Western blot,respectively.16S rDNA sequencing was used to analyze the structure of intestinal flora.Results Compared with the blank control group,the mice in the model group showed a decrease in body mass,an increase in DAI score,a decrease in colon length,serious mucosal injury and inflammatory cell infiltration,the contents of TNF-α,IL-1β,IL-6 and IL-8 in colon tissue significantly increased(P<0.05,P<0.01),the content of IL-10 decreased(P<0.01),the mRNA expressions of LC3 and Beclin-1 in colon tissue decreased(P<0.05,P<0.01),the mRNA and protein expression of p62 increased(P<0.01),while the expressions of LC3Ⅱ/LC3Ⅰ and Beclin-1 proteins decreased(P<0.05,P<0.01).Compared with the model group,the body mass of mice in Kuijie Ankang Decoction group and SASP group increased(P<0.05),DAI score decreased(P<0.05),the colon length increased(P<0.05),the pathological damage of colon mucosa was alleviated,the contents of TNF-α,IL-1β,IL-6 and IL-8 in colon tissue decreased(P<0.05,P<0.01),the content of IL-10 increased(P<0.01),the expressions of LC3 and Beclin-1 mRNA in colon tissue increased(P<0.05,P<0.01),the expression of p62 mRNA and protein decreased(P<0.05,P<0.01),the expressions of LC3Ⅱ/LC3Ⅰand Beclin-1 protein increased(P<0.05,P<0.01).16S rDNA sequencing results showed that the diversity and evenness of the intestinal flora in the model group mice decreased,with a decrease in the relative abundance of Firmicutes,Actinobacteria and Patescibacteria(P<0.05),and an increase in the relative abundance of Bacteroidota,Verrucomicrobiota and Proteobacteria(P<0.05);the relative abundance of Bacilli and Coriobacteriia decreased(P<0.05),the relative abundance of Bacteroidia,Clostridia and Verrucomicrobiae increased(P<0.05);the relative abundance of Ligilactobacillus and Dubosiella decreased(P<0.05),the relative abundance of unclassified Muribaculaceae,Lachnospiraceae NK4A136_group,Akkermansia and unclassified Lachnospiraceae increased(P<0.05).Compared with the model group,the diversity and evenness of intestinal flora increased in Kuijie Ankang Decoction group and SASP group,with an increase in the relative abundance of Firmicutes(P<0.05),a decrease in the relative abundance of Bacteroidota and Verrucomicrobiota(P<0.05),the relative abundance of Bacteroidia and Bacilli increased(P<0.05),the relative abundance of Verrucomicrobiae decreased(P<0.05);the relative abundance of unclassified Muribaculaceae and Ligilactobacillus increased(P<0.05),while the relative abundance of Lachnospiraceae NK4A136_group and Akkermansia decreased(P<0.05).Conclusion Kuijie Ankang Decoction can significantly improve the intestinal mucosal injury of UC mice,and the mechanism may be related to the regulation of colon autophagy level and intestinal flora disorder.

Objective:To explore the therapeutic effect of compound Kuijie Ankang Decoction on ulcerative colitis (UC) model mice by non targeted metabonomics; To explore its mechanism.Compound Kuijie Ankang.Methods:The mice were randomly divided into blank control group, model group, Kuijie Ankang Decoction group and sulfasalazine group, with 12 mice in each group. Except the blank control group, the other groups were given 1.5% DSS solution for free drinking to prepare UC model. After successful modeling, Kuijie Ankang Decoction group was intragastrically administered with compound Kuijie Ankang Decoction of 9.68 g/kg, sulfasalazine group was intragastrically administered with sulfasalazine capsule suspension of 320 mg/kg, model group and blank control group were intragastrically administered with equal volume of purified water, once a day, for 7 consecutive days. The body mass and disease activity index (DAI) score of mice were measured. ELISA was used to measure the levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6), and interleukin-10 (IL-10) in the colon tissue of mice; the protein expressions of Claudin-1 and Zo-1 in colon tissue were detected by immunofluorescence method. HE staining was used to observe the pathological changes in the colon, and UHPLC-OE-MS technology was used to analyze the endogenous metabolite structure of mouse colon tissue, differential metabolites and related metabolic pathways were screened.Results:Compared with the model group, the colon length in Kuijie Ankang Decoction group and sulfasalazine group increased ( P<0.01), the DAI score decreased ( P<0.01), the levels of TNF-α, IL-1β and IL-6 in colon tissue decreased ( P<0.01), the level of IL-10 increased ( P<0.01), and the average optical density of Claudin-1 and Zo-1 protein increased ( P<0.01 or P<0.05). Metabolomics analysis identified 26 potential differential metabolites, including nicotinamide adenine dinucleotide, guanine, gamma aminobutyric acid, and thiamine, affecting 26 key metabolic pathways, including lysine biosynthesis, thiamine metabolism, cysteine and methionine metabolism. Conclusion:Kuaijie Ankang Decoction may improve metabolites such as Gamma aminobutyric acid and thiamine through metabolic pathways such as lysine biosynthesis to alleviate inflammatory reactions, thereby exerting therapeutic effects on ulcerative colitis in mice.

Objective To investigate the mechanism of Kuijie Ankang Decoction in regulating immune,autophagy and intestinal flora in the treatment of ulcerative colitis(UC).Methods UC mouse model was established by free drinking with sodium dextran sulfate.The mice were randomly divided into blank control group,model group,Kuijie Ankang Decoction group,salazine sulfopyridine(SASP)group and 3-methyladenine(3-MA)group,with 12 mice in each group.Each drug group was given corresponding drugs for gavage,the blank control group and model group were given the same volume of distilled water for gavage for 7 days.The general condition of mice was observed and the disease activity index(DAI)was scored,the morphology of colon tissue was observed by HE staining,the contents of TNF-α,IL-1β,IL-6,IL-8 and IL-10 in colon tissue were detected by ELISA,the mRNA and protein expressions of LC3,Beclin-1 and p62 in colon tissue were detected by RT-qPCR and Western blot,respectively.16S rDNA sequencing was used to analyze the structure of intestinal flora.Results Compared with the blank control group,the mice in the model group showed a decrease in body mass,an increase in DAI score,a decrease in colon length,serious mucosal injury and inflammatory cell infiltration,the contents of TNF-α,IL-1β,IL-6 and IL-8 in colon tissue significantly increased(P<0.05,P<0.01),the content of IL-10 decreased(P<0.01),the mRNA expressions of LC3 and Beclin-1 in colon tissue decreased(P<0.05,P<0.01),the mRNA and protein expression of p62 increased(P<0.01),while the expressions of LC3Ⅱ/LC3Ⅰ and Beclin-1 proteins decreased(P<0.05,P<0.01).Compared with the model group,the body mass of mice in Kuijie Ankang Decoction group and SASP group increased(P<0.05),DAI score decreased(P<0.05),the colon length increased(P<0.05),the pathological damage of colon mucosa was alleviated,the contents of TNF-α,IL-1β,IL-6 and IL-8 in colon tissue decreased(P<0.05,P<0.01),the content of IL-10 increased(P<0.01),the expressions of LC3 and Beclin-1 mRNA in colon tissue increased(P<0.05,P<0.01),the expression of p62 mRNA and protein decreased(P<0.05,P<0.01),the expressions of LC3Ⅱ/LC3Ⅰand Beclin-1 protein increased(P<0.05,P<0.01).16S rDNA sequencing results showed that the diversity and evenness of the intestinal flora in the model group mice decreased,with a decrease in the relative abundance of Firmicutes,Actinobacteria and Patescibacteria(P<0.05),and an increase in the relative abundance of Bacteroidota,Verrucomicrobiota and Proteobacteria(P<0.05);the relative abundance of Bacilli and Coriobacteriia decreased(P<0.05),the relative abundance of Bacteroidia,Clostridia and Verrucomicrobiae increased(P<0.05);the relative abundance of Ligilactobacillus and Dubosiella decreased(P<0.05),the relative abundance of unclassified Muribaculaceae,Lachnospiraceae NK4A136_group,Akkermansia and unclassified Lachnospiraceae increased(P<0.05).Compared with the model group,the diversity and evenness of intestinal flora increased in Kuijie Ankang Decoction group and SASP group,with an increase in the relative abundance of Firmicutes(P<0.05),a decrease in the relative abundance of Bacteroidota and Verrucomicrobiota(P<0.05),the relative abundance of Bacteroidia and Bacilli increased(P<0.05),the relative abundance of Verrucomicrobiae decreased(P<0.05);the relative abundance of unclassified Muribaculaceae and Ligilactobacillus increased(P<0.05),while the relative abundance of Lachnospiraceae NK4A136_group and Akkermansia decreased(P<0.05).Conclusion Kuijie Ankang Decoction can significantly improve the intestinal mucosal injury of UC mice,and the mechanism may be related to the regulation of colon autophagy level and intestinal flora disorder.

Objective To investigate the effects of combined exercise training on exercise ability and inflammatory factors in elderly patients with chronic obstructive pulmonary disease(COPD)complicated with sarcopenia.Methods Sixty elderly patients with COPD complicated with sarcopenia who were treated in Hangzhou Third People's Hospital from July 2022 to May 2023 were selected as study objects,and were divided into control group and treatment group according to random number table method,with 30 cases in each group.The control group received routine treatment of COPD,while the treatment group received combined exercise training on the basis of routine treatment for 12 weeks.The appendicular skeletal muscle mass index(ASMI),grip strength,walking speed,6 min walking distance(6MWD),COPD assessment test(CAT),forced expiratory volume in one second(FEV1)/forced vital capacity(FVC)、serum interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)were compared.The correlation between serum IL-6、TNF-α and ASMI,grip strength、walking speed were analyzed.Results After treatment,6MWD in both groups was significantly longer than before treatment,and CAT score was significantly lower than before treatment(P＜0.05).The ASMI,grip strength and walking speed of patients in treatment group were significantly higher than before treatment,and the serum IL-6 and TNF-α were significantly lower than before treatment(P＜0.05).The ASMI,grip strength and walking speed of treatment group were significantly higher than those of control group,6MWD was significantly longer than that of control group,CAT score,serum IL-6 and TNF-α were significantly lower than those of control group(P＜0.05).There was no significant difference in FEV1/FVC between two groups before and after treatment(P＞0.05).Pearson correlation analysis showed that serum IL-6、TNF-α were negatively correlated with ASMI、grip strength and walking speed.Conclusion Combined exercise training can improve muscle mass and strength,improve exercise ability and clinical symptoms,and inhibit inflammatory response in elderly patients with COPD complicated with sarcopenia.

Objective:To evaluate the bone hook pulling technique in reduction of valgus-impacted femoral neck fractures.Methods:A retrospective study was conducted to analyze the clinical data of 12 patients who had been treated from September 2021 to September 2022 for valgus-impacted femoral neck fractures at Department of Orthopaedics and Traumatology, Honghui Hospital. There were 6 males and 6 females with an age of (43.8±10.1) years. By the Garden classification, the 12 cases were all type Ⅰ; by the Pauwels classification, 10 cases were type Ⅰ and 2 cases type Ⅱ. All patients were definitively diagnosed preoperatively by radiographs and CT images. The bone hook pulling technique was used to extend the bone hook from the lateral side to the medial side of the lesser trochanter, pull the bone hook to the lateral side, and reset the fracture with the help of pulling force and the action of the surrounding soft tissue hinge. The fractures were then fixed with the Femoral Neck System (FNS). Length of surgical incision, number of fluoroscopy for bone hook-related operations, operation time, quality of fracture reduction, fracture healing time, functional recovery of the hip and shortening of the femoral neck at the last follow-up were recorded.Results:The 12 valgus-impacted femoral neck fractures were successfully reset by the hook pulling technique. In this cohort, the length of surgical incision was (4.6±0.7) cm, the number of fluoroscopy for bone hook-related operations (4.3±0.7) times, and the operation time (54.3±4.2) min. The 12 patients were followed up for (11.5±4.2) months postoperatively and the fracture healing time (4.2±0.7) months. According to the Garden score, the quality of postoperative fracture reduction was assessed as grade Ⅰ in 10 cases and as grade Ⅱ in 2 cases. According to the Harris hip score, the hip function was assessed as excellent in 10 cases and as good in 2 cases at the last follow-up. The length of femoral neck shortening was (1.17±0.68) mm at the last follow-up in the 12 patients; no complications related to fracture reduction were observed.Conclusion:In the reduction of valgus-impacted femoral neck fractures, bone hook pulling technique shows advantages of operational simplicity, a high rate of successful reduction, and satisfactory clinical effects.

Objective:To evaluate the bone hook pulling technique in reduction of valgus-impacted femoral neck fractures.Methods:A retrospective study was conducted to analyze the clinical data of 12 patients who had been treated from September 2021 to September 2022 for valgus-impacted femoral neck fractures at Department of Orthopaedics and Traumatology, Honghui Hospital. There were 6 males and 6 females with an age of (43.8±10.1) years. By the Garden classification, the 12 cases were all type Ⅰ; by the Pauwels classification, 10 cases were type Ⅰ and 2 cases type Ⅱ. All patients were definitively diagnosed preoperatively by radiographs and CT images. The bone hook pulling technique was used to extend the bone hook from the lateral side to the medial side of the lesser trochanter, pull the bone hook to the lateral side, and reset the fracture with the help of pulling force and the action of the surrounding soft tissue hinge. The fractures were then fixed with the Femoral Neck System (FNS). Length of surgical incision, number of fluoroscopy for bone hook-related operations, operation time, quality of fracture reduction, fracture healing time, functional recovery of the hip and shortening of the femoral neck at the last follow-up were recorded.Results:The 12 valgus-impacted femoral neck fractures were successfully reset by the hook pulling technique. In this cohort, the length of surgical incision was (4.6±0.7) cm, the number of fluoroscopy for bone hook-related operations (4.3±0.7) times, and the operation time (54.3±4.2) min. The 12 patients were followed up for (11.5±4.2) months postoperatively and the fracture healing time (4.2±0.7) months. According to the Garden score, the quality of postoperative fracture reduction was assessed as grade Ⅰ in 10 cases and as grade Ⅱ in 2 cases. According to the Harris hip score, the hip function was assessed as excellent in 10 cases and as good in 2 cases at the last follow-up. The length of femoral neck shortening was (1.17±0.68) mm at the last follow-up in the 12 patients; no complications related to fracture reduction were observed.Conclusion:In the reduction of valgus-impacted femoral neck fractures, bone hook pulling technique shows advantages of operational simplicity, a high rate of successful reduction, and satisfactory clinical effects.

Objective:To observe any effect of fast walking on walking speed, 6-minute walking test (6MWT) time, and on serum levels of growth differentiation factor-8 (GDF-8) and insulin-like growth factor-1 (IGF-1) in patients with sarcopenia.Methods:A total of 61 sarcopenia patients were randomly divided into an observation group ( n=31) and a control group ( n=30). Both groups were given conventional drug therapy. In addition, the observation group underwent 30-minutes of walking at 100-120 steps/min (about 60% of maximum heart rate) three times a week for 12 weeks. The control group also walked, but at 70-90 steps/min (less than 50% of maximum heart rate). Grip strength, walking speed, 6MWT time, skeletal muscle index (ASMI) and serum GDF-8 and IGF-1 were compared before and after the intervention. Results:There were no significant differences in grip strength or ASMI between observation group and control group (comparing males with males and females with females) before the experiment. Afterward, grip strength and ASMI in the observation group had increased significantly on average. Both were then significantly higher than the control groups′ averages, which had not changed significantly. Average walking speed, 6MWT time and serum IGF-1 levels had improved significantly in both groups, but the observation group′s average improvement was significantly greater. A significant decrease the average serum GDF-8 level was observed in the observation group, but not in the control group.Conclusion:Fast walking can improve the walking of persons with sarcopenia, raise serum IGF-1 levels, and significantly reduce serum GDF-8.

Atherosclerosis (AS) is a chronic and progressive arterial disease. It is an important cause of the occurrence and development of cardiovascular and cerebrovascular diseases. With the development of Traditional Chinese Medicine (TCM), TCM has many advantages in the therapy of AS, with less adverse reactions. Studies have shown that TCM can resist AS, and the mechanism mainly belongs to regulating lipid metabolism, anti-lipid peroxidation, anti-inflammation, anticoagulation, and protecting the structure and function of vascular endothelial cells. The mechanism of TCM for AS is warranted to be studied systematically, and the chemical components need to be further clarified.

Objective:To construct a bivalent DNA vaccine against SARS-CoV-2 and influenza A virus H3N2 and to evaluate its immunogenicity in mice.Methods:The coding sequences for spike 1 (S1) protein of SARS-CoV-2 Beta variant and hemagglutinin (HA) of influenza A virus Cambodia (H3N2) strain were codon-optimized and synthesized. The two coding genes were ligated by the self-cleaving 2A peptide using over-lapping PCR to construct S1-2A-HA fragment, which was inserted into pVRC vector to construct the bivalent DNA vaccine, named as pVRC-S1-2A-HA. Indirect immunofluorescence assay (IFA) and Western blot were performed to detect the expression of S1 and HA proteins. BALB/c mice were immunized with pVRC-S1-2A-HA by intramuscular injection and electroporation. The humoral immune responses induced in mice were detected by indirect ELISA, pseudovirus neutralization assay and hemagglutination inhibition assay. Cellular immune responses were detected by IFN-γ ELISPOT, intracellular cytokine staining (ICS) and cytometric bead array (CBA).Results:The bivalent DNA vaccine pVRC-S1-2A-HA could express S1 and HA proteins in vitro. Specific cellular immune responses against S1 protein and specific IgG antibody against HA protein were significantly induced in mice with single-dose immunization. The antigen-specific immunity was significantly enhanced after booster immunization. The geometric mean titer (GMT) of specific IgG antibody increased to 3 251 for S1 protein and 45 407 for HA protein after two-dose immunization. Moreover, the S1-specific T cells increased to 1 238 SFC/10 6 cells. ICS results indicated that the booster vaccination induced CD4 + T and CD8 + T cells to produce IL-2, IFN-γ and TNF-α in mice. The secretion of various cytokines including IL-2, IL- 4, IL-6, IL-10 and IFN-γ in mouse splenocytes was induced after single-dose immunization. Conclusions:A bivalent DNA vaccine against SARS-CoV-2 and influenza A virus H3N2 was constructed and could induce S1- and HA-specific humoral and cellular immune responses in mice, suggesting the great potential of it for further development and application.

Objective:To explore the degradation property of novel magnesium alloy stents in Bama mini-pig models of carotid artery stenosis, and evaluate the feasibility of observing their dynamic and continuous process by high-resolution C-arm CT.Methods:Twelve Bama mini-pigs were selected; carotid artery stenosis models were established by large balloon over-dilation and high-fat and high-salt diet in Bama pigs; 24 weeks after that, self-made braided degradable magnesium alloy stents were inserted into the carotid artery stenosis models (confirmed by DSA) by balloon dilation. Degrees of stent patency and in-stent restenosis were examined by DSA immediately after procedure and on the 30 th, 60 th and 90 th d of procedure. Four experimental pigs were sacrificed on the 30 th, 60 th and 90 th d of procedure, respectively; the degradation property of the novel magnesium alloy stents was observed according to results of high-resolution C-arm CT in the inserted-stent areas and staining results of specimens in the stenosis areas, and stent imaging features during degradation were summarized. Results:Twelve Bama mini-pig models of carotid artery stenosis were established and 12 magnesium alloy stents were successfully inserted with a technical success rate of 100%. Both immediate postoperative and follow-up angiography showed patency of the vascular lumens without obvious in-stent restenosis. High-resolution C-arm CT and pathological examination showed homogeneous stent lumens and clear delineation of the stent meshes, with slightly degraded stent on the 30 th d of procedure; the stent lumen was blurred and some magnesium alloy wires were fractured with developed degradation of the stent on the 60 th d of procedure; and the stent meshes and stent strut could not be visualized due to severe degradation of the stent on the 90 th d of procedure. Conclusion:Magnesium alloy degradable stent is almost completely degraded within 90 th d of procedure in Bama mini-pig models of carotid artery stenosis, and high-resolution C-arm CT can be used to dynamically monitor the degradation of the stent in vivo.