Objective:To investigate the preventive and therapeutic effects and mechanisms of Dachaihu decoction(DCD)on dextran sodium sulfate(DSS)-induced ulcerative colitis(UC)and liver injury in mice,as well as the association between DCD benefits and gut microbiota modulation.Methods:Mice were treated with DCD(20.10 and 10.05 g/kg)for 2 weeks,with free access to drinking water containing 3%DSS in the second week to induce UC.Histopathological examination,RT-qPCR and 16S rRNA sequencing were used to investigate the effect of DCD on UC mice.Results:DCD pretreatment significantly alleviated weight loss,bloody diarrhea with mucus,histopathological abnormalities of the colon,and colon shortening in mice with DSS-induced UC.In addition,DCD pretreat-ment significantly upregulated the levels of Occludin,ZO-1,and MUC-2 in the colon and protected the intestinal barrier of mice.DCD pretreatment also alleviated inflammatory cell infiltration in the colon and the liver and significantly reduced the expression levels of the proinflammatory factors such as IL-1β,IL-6,TNF-α,iNOS,COX-2,and NLRP3,thereby exerting a protective effect against UC and liver injury.It should be noted that DCD corrected gut micro-biota imbalance in UC mice by enriching probiotic bacteria such as Lactobacillus and Bifidobacterium and reducing harmful bacteria such as Norank_f_Desulfovibrionaceae and Escherichia-Shigella.Conclusion:DCD can alleviate DSS-induced UC and exert a liver-protecting effect by protecting intestinal barrier,inhibiting inflam-mation,and regulating gut microbiota.

Arteriovenous malformations(AVMs)are a kind of high-flow vascular malformation.AVMs can be classified in many ways,including histo-embryological classification,hemodynamic classification,etc.At present,the two mainstream classification systems used to guide the embolization treatment of peripheral AVMs are proposed by Cho and Yakes respectively based on the angiographic morphology of the lesions.Interventional embolization is the first-line treatment for AVMs.Among the many embolization agents,absolute ethanol is a permanent liquid embolization agent.Absolute ethanol can directly destroy the vascular endothelial cells to achieve a good curative efficacy,therefore,it has been wildly used in the treatment of peripheral AVMs.Yakes classification combines the angiographic classification with absolute ethanol embolization therapy.During absolute ethanol treatment,close attention should be paid to the occurrence of complications such as elevated pulmonary artery pressure.Although there are challenges remaining in the treatment of AVMs,the rapid development of molecular genetics has made targeted drug adjunctive treatment for AVMs possible.Perhaps,the novel therapeutic mode of combination use of traditional therapy targeted drug may be able to make a breakthrough in the treatment of AVMs.

Objective:To investigate the effects of wza gene deletion in Klebsiella pneumoniae on capsule formation ability and bacteriophage sensitivity. Methods:The wza deletion mutant strain was constructed through a temperature-sensitive plasmid-mediated homologous recombination. The growth curves of W14 and Δ wza were detected by measuring the optical density OD 600. In order to analyze the effect of gene wza on bacterial capsule formation, wild-type strain W14 and Δ wza mutant strain were detected by transmission electron microscope, and their capsule contents were measured by quantifying the uronic acid contents. The plaque assay was used to detect bacterial sensitivity to bacteriophage in wild-type strain W14 and Δ wza mutant strain. The t test was used to compare whether there were differences in the contents of uronic acid in the capsules of wild-type strain W14 and Δ wza mutant strain. Results:The PCR results revealed that the Δ wza mutant strain was successfully constructed. Compared with wild-type strain W14, the growth curves of Δ wza on the solid plates demonstrated a slightly slower growth. However, no difference in growth was observed among wild-type strain W14 and Δ wza mutant strains in LB broth. The transmission electron microscope results showed that wza gene deletion resulted in the loss of capsule in bacteria. The uronic acid content assay suggested that the capsule content was significantly decreased in Δ wza mutant strain (45.963±2.795) μg/ml compared with wild-type strain W14 (138.800±5.201) μg/ml. There was a statistical difference between the two groups ( t=27.233, P<0.001). The plaque assay indicated that bacteria lost its sensitivity to bacteriophage when gene wza was deleted. Conclusion:Deletion of the wza gene impairs bacterial capsule formation ability and can affect bacterial sensitivity to bacteriophage phiW14.

Objective:To express and purify the phage depolymerase from hypervirulent Klebsiella pneumoniae (hv Kp) serotype K1 and validate its function. Methods:Phage that infected serotype K1-type hv Kp was isolated from hospital sewage. The biology and morphology of the phage were determined by plaque assay and transmission electron microscopy. The whole genome of the phage was sequenced by the Illumina HiSeq 2500 platform. The presence of depolymerase was determined by observing the plaque halo. Bioinformatic analysis and prokaryotic protein expression system were further used to predict and identify phage depolymerase. The depolymerase gene fragment was obtained by PCR and cloned into the pET28a expression vector, and the expression and purification of the depolymerase were completed in strain BL21. The depolymerase activities on the capsular polysaccharide of serotype K1-type hv Kp clinical isolates were detected by plaque assay and low-speed centrifugation assay. Results:A lytic phage (phiA2) that infected serotype K1-type hv Kp clinical isolate was isolated from hospital sewage. It was typical of the Caudovirales order and Autographiviridae family, and its whole genome was 43 526 bp in length and contained 51 coding domain sequences. The phage phiA2-derived depolymerase phiA2-dep was predicted, expressed and purified. The plaque assay and low-speed centrifugation assay indicated that the depolymerase phiA2-dep had good lytic activity on the capsular polysaccharide of serotype K1-type hv Kp clinical isolates. Conclusion:Depolymerase phiA2-dep can specifically degrade the capsular polysaccharide of serotype K1-type hv Kp, which has potential application value in treating bacterial infection.

Objective:To investigate the effects of intra-articular injection of exosomes derived from dental pulp stem cells from hu-man exfoliated deciduous teeth(SHED-EXO)on subchondral bone homeostasis in rat temporomandibular joint osteoarthritis(TMJ OA)process.Methods:36 male SD rats were randomly divided into 3 groups(n=12):control(CON),sodium iodoacetate(MIA)-induced TMJ OA(MIA),and SHED-EXO injection into TMJ OA(SHED-EXO)groups.At 2 and 6 weeks post-treatment,Micro-CT,Double labeling,TRAP staining,and immunohistochemical staining were employed to detect osteoclasts and osteoblasts in the subchondral bone.Additionally,the mRNA expression levels of ADAMTs5,IL-1β,OCN and OPG/RANKL were analyzed by qRT-PCR.Results:The MIA group exhibited significant bone loss and an enlarged bone marrow cavity.In comparison with the CON group,BV/TV and Tb.Th were lower(P＜0.001),while BS/BV,Tb.Sp,and Tb.N were higher(P＜0.01).Additionally,the bone formation rate within 5 days was low-er than that of the control group(P＜0.001).When compared to the MIA group,the SHED-EXO group showed a significant increase in bone morphology and bone mass.BV/TV and Tb.Th were increased(P＜0.01),while BS/BV,Tb.Sp and Tb.N were decreased(P＜0.05).The bone formation rate was higher(P＜0.01).Compared with both the control and treatment groups,the MIA group exhibited a significant increase in the number of osteoclasts in the subchondral bone(P＜0.01),along with a notable decrease in H-type blood vessels and OCN-positive areas(P＜0.01).Conclusion:Intra-articular injection of SHED-EXO can reg-ulate condylar subchondral bone homeostasis in TMJ OA of rats by promoting osteogenesis and inhibiting osteoclasts.

Objective:To evaluate the feasibility of using two-dimensional shear wave elastography (2D-SWE) based liver and spleen elastic hardness (L/S-SWE) in patients with liver cirrhosis, and to determine the exclusion and diagnostic thresholds for early identification of liver cirrhosis.Methods:A total of 574 patients with chronic hepatitis B (hepatitis B for short) were included in this study. The clinical characteristics, L-SWE and S-SWE of the patients were collected, and the differences between cirrhosis group ( n=311) and non cirrhosis group ( n=263) were analyzed. The success rate and stability of liver and spleen elastic surgery were evaluated in two groups. The receiver operating characteristic (ROC) curve was used to analyze the efficacy of L-SWE, S-SWE, aspartate aminotransferase to platelet ratio index (APRI) alone and in combination in diagnosing liver cirrhosis. By analyzing the ROC curve, the double threshold for excluding and diagnosing liver cirrhosis was determined. Results:There was a statistically significant difference in platelet count and APRI between the cirrhosis group and the non cirrhosis group (all P<0.05). In the feasibility assessment of 2D-SWE technology, the success rate and stability of liver and spleen elastic operation were relatively high (success rate: 97.2% vs 81.3%; stability: 0.92 vs 0.84), and the success rate and stability of L-SWE operation were slightly better than S-SWE. The success rate of S-SWE operation in the cirrhosis group was higher than that in the non cirrhosis group ( P<0.05). The correlation analysis results showed that L-SWE, S-SWE, APRI were positively correlated with liver tissue pathological grading ( r=0.677, 0.528, 0.149, all P<0.05). The areas under the ROC curve for identifying liver cirrhosis using L-SWE, S-SWE, and APRI were 0.959, 0.896, and 0.706, respectively. When L-SWE and S-SWE were combined, the area under the ROC curve was 0.987, the sensitivity was 92.6%, and the specificity was 96.0%. The Delong test showed that the combined diagnosis of L-SWE and S-SWE had the same diagnostic efficacy as using L-SWE alone for liver cirrhosis ( P>0.05). Further analysis of the ROC curve showed that the likelihood of liver cirrhosis was low when L-SWE was less than 9.4 kPa, and high when L-SWE was greater than 12.0 kPa. Patients between 9.4 and 12.0 kPa can undergo further S-SWE testing; If the S-SWE was between 17.5 and 29.3 kPa, it was classified as 2D-SWE, which was difficult to determine whether there was liver cirrhosis, and further liver puncture and other examinations were needed. Conclusions:2D-SWE technology has high operational feasibility in the diagnosis of liver cirrhosis, and combined with S-SWE, it helps to improve the diagnostic efficiency of early non-invasive identification of liver cirrhosis, enabling more patients to avoid unnecessary liver puncture examinations.

Objective This study aimed to investigate the factors influencing the personal burden rate incerebral ische-mic patients,compare the difference in the burden rate among the patients with varying degrees of cerebral ischemia,provide a reference for establishing a personal burden rate evaluation,and propose suggestions for control its increase.Methods The medi-cal insurance data were collected from 8164 discharged patients in a tertiary hospital in Tianjin between January and December 2022.With the data,the Generalized Linear Model was utilized to analyze the factors affecting the personal burden rate across different Diagnosis Related Groups(DRGs).Results Statistically significant differences were observed in the cost structure a-mong different DRGs.Age,length of hospital stays,total hospitalization cost,hospital admission mode,number of hospitaliza-tions,and type of medical insurance significantly impacted the personal burden rate.The personal burden rate was inversely cor-related with age and length of hospital stays,but directly correlated with the total hospitalization cost.The patients admitted from emergency,first-time hospitalization,and those covered by the basic medical insurance program for urban employees had a lower personal burden rate.Conclusion Hospitals should establish diverse personal burden rate performance evaluation standards for patients with different types of medical insurance,incorporating factors such as average length of hospital stays and average hospi-talization cost.A more equitable hospital internal assessment plan should be developed by considering patients admitted to differ-ent departments and aligning with the characteristics of clinical pathways.Medical institutions should minimize self-funded pro-jects under declared medical insurance,increase the enrollment of cases in DRGs,and promote tiered diagnosis and treatment to reduce the personal burden rate for patients.

Objective:To investigate the effects of wza gene deletion in Klebsiella pneumoniae on capsule formation ability and bacteriophage sensitivity. Methods:The wza deletion mutant strain was constructed through a temperature-sensitive plasmid-mediated homologous recombination. The growth curves of W14 and Δ wza were detected by measuring the optical density OD 600. In order to analyze the effect of gene wza on bacterial capsule formation, wild-type strain W14 and Δ wza mutant strain were detected by transmission electron microscope, and their capsule contents were measured by quantifying the uronic acid contents. The plaque assay was used to detect bacterial sensitivity to bacteriophage in wild-type strain W14 and Δ wza mutant strain. The t test was used to compare whether there were differences in the contents of uronic acid in the capsules of wild-type strain W14 and Δ wza mutant strain. Results:The PCR results revealed that the Δ wza mutant strain was successfully constructed. Compared with wild-type strain W14, the growth curves of Δ wza on the solid plates demonstrated a slightly slower growth. However, no difference in growth was observed among wild-type strain W14 and Δ wza mutant strains in LB broth. The transmission electron microscope results showed that wza gene deletion resulted in the loss of capsule in bacteria. The uronic acid content assay suggested that the capsule content was significantly decreased in Δ wza mutant strain (45.963±2.795) μg/ml compared with wild-type strain W14 (138.800±5.201) μg/ml. There was a statistical difference between the two groups ( t=27.233, P<0.001). The plaque assay indicated that bacteria lost its sensitivity to bacteriophage when gene wza was deleted. Conclusion:Deletion of the wza gene impairs bacterial capsule formation ability and can affect bacterial sensitivity to bacteriophage phiW14.

Objective:To express and purify the phage depolymerase from hypervirulent Klebsiella pneumoniae (hv Kp) serotype K1 and validate its function. Methods:Phage that infected serotype K1-type hv Kp was isolated from hospital sewage. The biology and morphology of the phage were determined by plaque assay and transmission electron microscopy. The whole genome of the phage was sequenced by the Illumina HiSeq 2500 platform. The presence of depolymerase was determined by observing the plaque halo. Bioinformatic analysis and prokaryotic protein expression system were further used to predict and identify phage depolymerase. The depolymerase gene fragment was obtained by PCR and cloned into the pET28a expression vector, and the expression and purification of the depolymerase were completed in strain BL21. The depolymerase activities on the capsular polysaccharide of serotype K1-type hv Kp clinical isolates were detected by plaque assay and low-speed centrifugation assay. Results:A lytic phage (phiA2) that infected serotype K1-type hv Kp clinical isolate was isolated from hospital sewage. It was typical of the Caudovirales order and Autographiviridae family, and its whole genome was 43 526 bp in length and contained 51 coding domain sequences. The phage phiA2-derived depolymerase phiA2-dep was predicted, expressed and purified. The plaque assay and low-speed centrifugation assay indicated that the depolymerase phiA2-dep had good lytic activity on the capsular polysaccharide of serotype K1-type hv Kp clinical isolates. Conclusion:Depolymerase phiA2-dep can specifically degrade the capsular polysaccharide of serotype K1-type hv Kp, which has potential application value in treating bacterial infection.

Objective To investigate the expression of serum Sestrin2 and Toll-like receptor 7(TLR7)in patients with severe pneumonia and sepsis and their predictive value for heart failure.Methods A total of 86 patients with severe pneumonia and sepsis complicated with heart failure(complicated with heart failure group)and 86 patients with severe pneumonia and sepsis(uncomplicated with heart failure group)who were diagnosed and treated in Shanxi Provincial People's Hospital from February 2022 to May 2023 were studied.Another 86 patients who underwent health examinations were included as the control group.Enzyme linked immunosorbent assay(ELISA)method was applied to measure serum levels of Sestrin2 and TLR7.Color doppler echocardiography was applied to measure cardiac function related indicators:left ventricular ejection fraction(LVEF),left ventricular end diastolic diameter(LVEDD),and left ventricular end systolic diameter(LESD).The expression of Sestrin2 and TLR7 levels and cardiac function in three groups was analyzed.Pearson correlation was applied to analyze the relationship between serum levels of Sestrin2 and TLR7 and cardiac function related indicators in patients with heart failure.Receiver operating characteristic(ROC)curve was applied to analyze the predictive value of serum levels of Sestrin2 and TLR7 in patients with severe pneumonia and sepsis complicated with heart failure.Results The serum levels of Sestrin2(11.59±3.31 ng/ml,16.13±3.62 ng/ml),TLR7(48.93±9.52 ng/ml,61.74±10.11 ng/ml),and cardiac function related indicators[LVEDD(53.28±5.76 mm,62.54±6.11 mm)and LESD(38.16±4.38 mm,48.15±5.02 mm)]were higher in uncomplicated heart failure group and complicated heart failure group than those in control group(7.11±2.34 ng/ml,40.12±10.16 ng/ml,44.86±5.02 mm,29.02±4.07 mm),with significant differences(qSestrin2=13.241,26.659,qTLR7=8.224,20.182,qLVEDD=13.824,29.028,qLESD=18.805,39.359,all P＜0.05),and those values in complicated with heart failure group were higher than those in uncomplicated with heart failure group,with significant differences(q=13.418,11.985,15.203,20.554,all P＜0.05).The LVEF(55.43％±6.62％,41.67％±5.84％)index in uncomplicated heart failure group and complicated heart failure group was lower than that in control group(62.75％±7.16％),with significant differences(q=10.344,29.789,all P＜0.05),and the index in complicated with heart failure group was lower than that in uncomplicated with heart failure group,with significant difference(q=19.455,P＜0.05).Pearson correlation analysis showed that serum levels of Sestrin2 and TLR7 in patients with heart failure were negatively correlated with LVEF(r=-0.419,-0.467,all P＜0.05),and positively correlated with LVEDD and LESD(r=0.456,0.419;0.402,0.437,all P＜0.05).Sestrin2 was positively correlated with TLR7(r=0.641,P＜0.05).ROC curve results showed that the area under the curve(AUC)of serum Sestrin2 combined TLR7 for predicting heart failure in severe pneumonia sepsis patients was 0.940,with sensitivity of 74.9％and specificity of 73.3％.Conclusion The serum levels of Sestrin2 and TLR7 in patients with severe pneumonia and sepsis were elevated and may have good predictive value for patients with heart failure.