Celastrol and wilforlide A are the main active triterpenoids of the traditional Chinese medicine Lei Gong Teng, which have anti-tumour, anti-inflammatory and immunosuppressive activities, and are the material basis for the clinical efficacy of Lei Gong Teng-related Chinese medicinal preparations. By analysing the biosynthetic pathway of active ingredients, optimizing genetic elements and utilizing "cell factory" to produce triterpenoids heterologously will be an effective way to obtain from Tripterygium wilfordii in the future in a "low-cost and high-efficient" manner. CYP712Ks are the first cytochrome P450s involved in the skeleton modification of friedelane-type and oleanane-type triterpenoids in T. wilfordii, and they can catalyse the generation of polpunonic acid from friedelin and 3-epi-katonic acid from β-amyrin by carboxylation at C-29 position. In this study, four multifunctional TwCYP712K1/2/3/5 were used to clarify the catalytic function and substrate selection preference using in vivo functional characterization in Saccharomyces cerevisiae. The spatial structure of the protein-substrate binding was clarified through homology modeling and molecular docking, and then the differential amino acids in the active pocket of the protein were mutated to clarify the crucial amino acids determining the catalytic function and the selection of substrate structure. A total of 63 mutant elements were constructed, and the amino acid sites affecting the carboxylation function of TwCYP712Ks were analyzed. In particular, the key amino acids affecting the substrate selectivity of TwCYP712K2 towards oleanane-type and friedelane-type triterpenoids were revealed and the TwCYP712K2^F127I and TwCYP712K2^A227T mutants would result in a reversal of the product ratio. In conclusion, four proteins of TwCYP712Ks were semi-rationally designed by homologous protein alignment and mutual mutation to elucidate multiple amino acid sites determining the catalytic function of the proteins, and a series of activity-enhancing or altering mutants were obtained, which provide abundant catalytic elements for the biosynthesis of active triterpenoids from T. wilfordii, and the mechanism of carboxylation in the C-29 position was initially elucidated.

OBJECTIVE To optimize the deproteinization process of Isatidis Radix polysaccharides and further explore its immu-nomodulatory activity,and to provide a scientific basis for the development and utilization of it.METHODS The optimum conditions of enzymatic deproteinization were optimized by a single factor combined with the Box-Behnken response surface method.The chemical composition and structural characteristics of deproteinized Isatidis Radix polysaccharides were analyzed by UV-visible spectrum,Fourier transform-infrared spectroscopy,high-performance gel permeation chromatography,high-performance liquid chromatography and scan-ning electron microscopy.The effects of deproteinized Isatidis Radix Polysaccharide on neutrophils,macrophages,IL-1β and IL-6 in zebrafish were investigated by using a zebrafish immunocompromised model.RESULTS The optimal enzymatic deproteinization process was as follows:trypsin 500 U·mL-1,pH 8.0,enzymatic hydrolysis time 5 h,enzymatic hydrolysis temperature 37℃.The deproteinization rate was(86.39±0.07)%,and the comprehensive score was(91.15±0.37)%.Ultraviolet,infrared spectroscopy scanning and scanning electron microscopy showed that the protein contained in the crude polysaccharide could be removed by enzymat-ic method.The relative molecular weight of the polysaccharides were between 5.82 and 60.26 kDa.The monosaccharide mole compo-sition was mannose ∶ rhamnose ∶ galacturonic acid ∶ glucose ∶ galactose ∶ arabinose=2.17 ∶ 0.96 ∶ 2.90 ∶ 83.25 ∶ 4.88 ∶ 5.84.The results of immune activity evaluation showed that when the concentration of deproteinized Radix Isatidis polysaccharides was 50～300 μg·mL-1,it could significantly increase the density of zebrafish immune cells,increase the number of macrophages,and reduce the content of IL-1β and IL-6 in immunocompromised zebrafish,thus exerting immunomodulatory effects.CONCLUAION The enzy-matic method can effectively remove the proteins contained in the crude polysaccharides of Isatidis Radix,and the deproteinized Isatidis Radix polysaccharides have certain immunomodulatory effects.

Objective To monitor the susceptibility of clinical isolates to antimicrobial agents in tertiary hospitals in major regions of China in 2022.Methods Clinical isolates from 58 hospitals in China were tested for antimicrobial susceptibility using a unified protocol based on disc diffusion method or automated testing systems.Results were interpreted using the 2022 Clinical &Laboratory Standards Institute(CLSI)breakpoints.Results A total of 318 013 clinical isolates were collected from January 1,2022 to December 31,2022,of which 29.5％were gram-positive and 70.5％were gram-negative.The prevalence of methicillin-resistant strains in Staphylococcus aureus,Staphylococcus epidermidis and other coagulase-negative Staphylococcus species(excluding Staphylococcus pseudintermedius and Staphylococcus schleiferi)was 28.3％,76.7％and 77.9％,respectively.Overall,94.0％of MRSA strains were susceptible to trimethoprim-sulfamethoxazole and 90.8％of MRSE strains were susceptible to rifampicin.No vancomycin-resistant strains were found.Enterococcus faecalis showed significantly lower resistance rates to most antimicrobial agents tested than Enterococcus faecium.A few vancomycin-resistant strains were identified in both E.faecalis and E.faecium.The prevalence of penicillin-susceptible Streptococcus pneumoniae was 94.2％in the isolates from children and 95.7％in the isolates from adults.The resistance rate to carbapenems was lower than 13.1％in most Enterobacterales species except for Klebsiella,21.7％-23.1％of which were resistant to carbapenems.Most Enterobacterales isolates were highly susceptible to tigecycline,colistin and polymyxin B,with resistance rates ranging from 0.1％to 13.3％.The prevalence of meropenem-resistant strains decreased from 23.5％in 2019 to 18.0％in 2022 in Pseudomonas aeruginosa,and decreased from 79.0％in 2019 to 72.5％in 2022 in Acinetobacter baumannii.Conclusions The resistance of clinical isolates to the commonly used antimicrobial agents is still increasing in tertiary hospitals.However,the prevalence of important carbapenem-resistant organisms such as carbapenem-resistant K.pneumoniae,P.aeruginosa,and A.baumannii showed a downward trend in recent years.This finding suggests that the strategy of combining antimicrobial resistance surveillance with multidisciplinary concerted action works well in curbing the spread of resistant bacteria.

Objective To investigate the changing distribution and antibiotic resistance profiles of clinical isolates of Staphylococcus in hospitals across China from 2015 to 2021.Methods Antimicrobial susceptibility testing was conducted for the clinical isolates of Staphylococcus according to the unified protocol of CHINET(China Antimicrobial Surveillance Network)using disk diffusion method and commercial automated systems.The CHINET antimicrobial resistance surveillance data from 2015 to 2021 were interpreted according to the 2021 CLSI breakpoints and analyzed using WHONET 5.6.Results During the period from 2015 to 2021,a total of 204,771 nonduplicate strains of Staphylococcus were isolated,including 136,731(66.8％)strains of Staphylococcus aureus and 68,040(33.2％)strains of coagulase-negative Staphylococcus(CNS).The proportions of S.aureus isolates and CNS isolates did not show significant change.S.aureus strains were mainly isolated from respiratory specimens(38.9±5.1)％,wound,pus and secretions(33.6±4.2)％,and blood(11.9±1.5)％.The CNS strains were predominantly isolated from blood(73.6±4.2)％,cerebrospinal fluid(12.1±2.5)％,and pleural effusion and ascites(8.4±2.1)％.S.aureus strains were mainly isolated from the patients in ICU(17.0±7.3)％,outpatient and emergency(11.6±1.7)％,and department of surgery(11.2±0.9)％,whereas CNS strains were primarily isolated from the patients in ICU(32.2±9.7)％,outpatient and emergency(12.8±4.7)％,and department of internal medicine(11.2±1.9)％.The prevalence of methicillin-resistant strains was 32.9％in S.aureus(MRSA)and 74.1％in CNS(MRCNS).Over the 7-year period,the prevalence of MRSA decreased from 42.1％to 29.2％,and the prevalence of MRCNS decreased from 82.1％to 68.2％.MRSA showed higher resistance rates to all the antimicrobial agents tested except trimethoprim-sulfamethoxazole than methicillin-susceptible S.aureus(MSSA).Over the 7-year period,MRSA strains showed decreasing resistance rates to gentamicin,rifampicin,and levofloxacin,MRCNS showed decreasing resistance rates to gentamicin,erythromycin,rifampicin,and trimethoprim-sulfamethoxazole,but increasing resistance rate to levofloxacin.No vancomycin-resistant strains were detected.The prevalence of linezolid-resistant MRCNS increased from 0.2％to 2.3％over the 7-year period.Conclusions Staphylococcus remains the major pathogen among gram-positive bacteria.MRSA and MRCNS were still the principal antibiotic-resistant gram-positive bacteria.No S.aureus isolates were found resistant to vancomycin or linezolid,but linezolid-resistant strains have been detected in MRCNS isolates,which is an issue of concern.

Hip fracture in the elderly is characterized by high incidence, high disability rate, and high mortality and has been recognized as a public health issue threatening their health. Surgery is the preferred choice for the treatment of elderly patients with hip fracture. However, lower extremity deep venous thrombosis (DVT) has an extremely high incidence rate during the perioperative period, and may significantly increase the risk of patients′ death once it progresses to pulmonary embolism. In response to this issue, the clinical guidelines and expert consensuses all emphasize active application of comprehensive preventive measures, including basic prevention, physical prevention, and pharmacological prevention. In this prevention system, basic prevention is the basis of physical and pharmacological prevention. However,there is a lack of unified and definite recommendations for basic preventive measures in clinical practice. To this end, the Orthopedic Nursing Professional Committee of the Chinese Nursing Association and Nursing Department of the Orthopedic Branch of the China International Exchange and Promotive Association for Medical and Health Care organized relevant nursing experts to formulate Expert consensus on perioperative basic prevention for lower extremity deep venous thrombosis in elderly patients with hip fracture ( version 2024) . A total of 10 recommendations were proposed, aiming to standardize the basic preventive measures for lower extremity DVT in elderly patients with hip fractures during the perioperative period and promote their subsequent rehabilitation.

Objective: To analyze and compare therapy responses, outcomes, and incidence of severe hematologic adverse events of flumatinib and imatinib in patients newly diagnosed with chronic phase chronic myeloid leukemia (CML) . Methods: Data of patients with chronic phase CML diagnosed between January 2006 and November 2022 from 76 centers, aged ≥18 years, and received initial flumatinib or imatinib therapy within 6 months after diagnosis in China were retrospectively interrogated. Propensity score matching (PSM) analysis was performed to reduce the bias of the initial TKI selection, and the therapy responses and outcomes of patients receiving initial flumatinib or imatinib therapy were compared. Results: A total of 4 833 adult patients with CML receiving initial imatinib (n=4 380) or flumatinib (n=453) therapy were included in the study. In the imatinib cohort, the median follow-up time was 54 [interquartile range (IQR), 31-85] months, and the 7-year cumulative incidences of CCyR, MMR, MR(4), and MR(4.5) were 95.2%, 88.4%, 78.3%, and 63.0%, respectively. The 7-year FFS, PFS, and OS rates were 71.8%, 93.0%, and 96.9%, respectively. With the median follow-up of 18 (IQR, 13-25) months in the flumatinib cohort, the 2-year cumulative incidences of CCyR, MMR, MR(4), and MR(4.5) were 95.4%, 86.5%, 58.4%, and 46.6%, respectively. The 2-year FFS, PFS, and OS rates were 80.1%, 95.0%, and 99.5%, respectively. The PSM analysis indicated that patients receiving initial flumatinib therapy had significantly higher cumulative incidences of CCyR, MMR, MR(4), and MR(4.5) and higher probabilities of FFS than those receiving the initial imatinib therapy (all P<0.001), whereas the PFS (P=0.230) and OS (P=0.268) were comparable between the two cohorts. The incidence of severe hematologic adverse events (grade≥Ⅲ) was comparable in the two cohorts. Conclusion: Patients receiving initial flumatinib therapy had higher cumulative incidences of therapy responses and higher probability of FFS than those receiving initial imatinib therapy, whereas the incidence of severe hematologic adverse events was comparable between the two cohorts.
Adult ; Humans ; Adolescent ; Imatinib Mesylate/adverse effects* ; Incidence ; Antineoplastic Agents/adverse effects* ; Retrospective Studies ; Pyrimidines/adverse effects* ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive/drug therapy* ; Treatment Outcome ; Benzamides/adverse effects* ; Leukemia, Myeloid, Chronic-Phase/drug therapy* ; Aminopyridines/therapeutic use* ; Protein Kinase Inhibitors/therapeutic use*

Objective To observe the effects of 4-week low intensity treadmill exercise on the learning and memory, amino acid levels and the protein expression of protein kinase A ( PKA) , cyclic adenosine monophosphate response element binding protein( CREB) and brain-derived neurotrophic factor(BDNF) in the prefrontal cortex (PFC) of the vascular dementia (VD) rats. Methods Thirty-nine SD rats were randomly allocated to 3 groups, sham group (sham, n= 13) , vascular dementia group (VD, n= 13) and vascular dementia treaded with exercise group (VD + EX, n= 13). Chronic cerebral ischemia model in VD group and VD+EX group rats were established by permanent ligation of bilateral, then VD+EX group rats were submitted to 4-week low intensity treadmill exercise. After exercise, spatial learning and memory ability were evaluated by Moms water maze test ( MWM ) , glutamic ( Glu ) and gamma-aminobutyric acid (GABA) levels in the PFC were measured by high performance liquid chromatography( HPLC) ; the protein expression of PKA, CREB and BDNF in the PFC of rats were detected by Western blotting. Results The result of the MWM showed the average escape latency of rats in the VD group on the 1 -5 days was significantly higer than sham group, the time to first find the original platform was significantly prolonged and the platform crossings decreased significantly ( P < 0 . 01 ) . The average escape latency of rats treated with four-week low intensity treadmill exercise on the 1-5 days was significantly lower than VD group, the time to first find the original platform was significantly shortened and the platform crossings increased significantly(P<0. Of). HPLC and Western blotting detection showed that the levels of Glu and GABA and the protein expression of PKA, CREB and BDNF in the PFC decreased significantly in VD group rats when compared with sham group ( P < 0.01 ) . Glu level and the protein expression of PKA, CREB and BDNF in the PFC of rats increased significantly ( P < 0 . 05) in VD + EX group rats when compared with VD group rats, but GABA level was not significant difference ( P > 0. 05 ) between the two groups. Conclusion Four-week low-intensity running exercise improves the learning and memory ability of VD rats through enhancing the Glu level and activating PKA-CREB-BDNF signaling in the PFC of rats.

Objective To examine the effect of estradiol (E2) treatment on ovariectomy (OVX) induced depressive-like behavior and possible mechanism by measuring inflammatory biomarkers levels oi interleukin-6 (IL-6) and tumor necrosis tactor-(x (TNF-ot) and brain-derived neurotrophic factor (BDNF) expression in amygdaia nucleus. Methods Thirty nine healthy aduit 缶male SD rats were randomly divided into three groups : sham operation group (SO), ovariectomized group (OVX) and ovariectomized estradiol treatment group (OVX + E2). After 6 weeks of E2 treatment, depressive like behavior was evaluated by opening field test (OFT) and sugar water preference test (SPT). The levels of inflammatory factors IL-6 and TNF-a in amygdala were measured by ELESA, and the expression of BDNF in rat amygdala was detected by immunohistochemical staining. Results The result of the SPT showed that OVX significantly decreased the sugar intake and sugar preference rate of rats, and E2 treatment significantly increased sugar water intake and sugar water preference rate of rats. The result of the OFT showed that OVX significantly decreased the numbers of crossing and rearing of rats, and reduced the time spent in the centre ; E2 treatment significantly increased the numbers of crossing and rearing of rats, and prolonged the time spent in the centre. ELESA and immunohistochemical analysis found the levels of IL-6 and TNF-a in amygdala increased significantly, while the average absorbance (AA) of BDNF in the amygdala reduced significantly (P<0.01 respectively) of rats in OVX group when compared with the SO group. And the levels of IL-6 and TNF-a in amygdala decreased significantly (P<0.01 respectively), while the A4 of BDNF increased significantly (P< 0.05) in the amygdala of rats in the OVX+EX group when compared with the OVX group. The difference was statistically significant. Conclusion E2 treatment improved depression-like behavior of OVX rats is partly due to increased antiinflammatory and activated the BDNF expression in amygdaloid nucleus, thus enhancing the neuroprotective effect of OVX rats.

Objective To study the effect and possible mechanism of thioridazine on the biological behavior of liver cancer Huh-7 cells.Methods Hepatocellular carcinoma Huh-7 cells were randomly divided into a control group and low,medium,and high concentration groups of thiazidine.The miR-3174 mimics and its negative control,miR-3174 inhibitor and its negative control were transfectel to Huh-7 cells,and the miR-3174 mimics and its negative control were treated with thiolidazine.Effect of thioridazine on the proliferation of Huh-7 cells was assessed by CCK-8 method and plate cloning experiment;effect of thioridazine on the migration and invasion of Huh-7 cells was calculated using scratch healing test and Transwell experiment;effect of thioridazine on the expression of miR-3174 in Huh-7 cells was detected by RT-qPCR;E-cadherin and N-cadherin proteins expression was analyzed using Western blotting.The effects of interference with miR-3174 or miR-3174 up-regulation combined with thioridazine on the biological behavior of Huh-7 cells were detected by the above methods.Results After thiolidazine treatment and interference with miR-3174 expression,miR-3174 expression in Huh-7 cells was decreased,proliferation,migration and invasion ability,N-cadherin protein expression were decreased,and E-cadherin protein expression was increased(P＜0.05).Up-regulating the expression of miR-3174 attenuated the effect of thiolidazine treatment on the malignant biological behavior of Huh-7 cells.Conclusion Thioridazine inhibits the proliferation,migration and invasion of liver cancer Huh-7 cells by down-regulating miR-3174 expression.

Objective To investigate the clinical efficacy and application value of traditional Chinese medicine Shugan Jiangzhi decoction combined with L-ornithine-L-aspartate(LOLA)in the treatment of fatty liver after viral hepatitis.Methods Patients with fatty liver after viral hepatitis who were diagnosed and treated in Hebei Chest Hospital from October 2018 to October 2020 were enrolled and randomly divided into control group and test group.The control group was treated with LOLA,and the test group was treated with Shugan Jiangzhi decoction on the basis of the treatment of the control group,and the changes of liver function,blood lipids and immune function before and after treatment in the two groups were observed and compared.Results A total of 144 patients were included in the study,including 72 patients in the control group and 72 cases in the test group.After treatment,the total effective rate of the test group was 93.06% ,which was higher than that of the control group(79.17% )(P<0.05).After treatment,the serum levels of ALT,AST,TBIL,DBIL,GGT,TC,LDL-C and TG in the two groups were significantly decreased(P<0.05),and the test group was lower than that in the control group(P<0.05).The ratio of HDL-C value to CD4+ T lymphocytes in the two groups was higher than that before treatment(P<0.05),and the test group was higher than that in the control group(P<0.05),but there was no significant change in the proportion of CD8+T lymphocytes(P>0.05).The ultrasound grade of fatty liver in the test group was better than that in the control group(P<0.05).Conclusion Shugan Jiangzhi decoction combined with LOLA can improve the immune function and the liver function,balance the blood lipid level,and continuously improve the prognosis of patients with fatty liver after viral hepatitis.