Objective To investigate the clinical efficacy and safety of simultaneous treatment of small hepatocellular carcinoma(HCC)with transarterial chemoembolization(TACE)combined with cone-beam computed tomography-guided(CBCT-guided)microwave ablation(MWA).Methods The clinical data of 69 patients with small HCC(72 lesions in total),who underwent TACE combined with CBCT-guided MWA simultaneous treatment from March 2018 to December 2022 at First Affiliated Hospital of Soochow University hospital,were retrospectively analyzed.Follow-up check was performed at 1,3,6,and 12 months after treatment.The mRECIST criteria was used to evaluate the tumor response.The objective response rate(ORR),disease control rate(DCR),progression-free survival(PFS),local tumor progression(LTP),and adverse reactions were analyzed.Results The initial complete remission(CR)rate,ORR and DCR of TACE combined with CBCT-guided MWA simultaneous treatment for small HCC was 94.2％(65/69),100％and 100％respectively.44.9％(31/69)of patients experienced tumor progression,and 20.3％(14/69)of patients experienced local tumor progression.Univariate and multivariate analyses showed that the maximum tumor diameter(≥2 cm and＜3 cm)was the main risk factor for PFS(HR=4.498,P＜0.001).No serious adverse events occurred during the study.Conclusion TACE combined with CBCT-guided MWA simultaneous treatment for small HCC is clinically effect and safe,and this therapy is particularly suitable for the treatment of lesions where the use of traditional image-guided methods is limited.

Objective To observe the clinical effect of lumbar fixed-point pulling method combined with magnetic needling in the treatment of lumbar disc herniation(LDH).Methods Sixty LDH patients were randomly divided into treatment group and control group,30 cases in each group.The treatment group was treated with lumbar fixed-point pulling method combined with magnetic needling,and the control group was treated with Celecoxib Capsules.Two weeks as a course of treatment and both groups were treated for two courses.After 1 month of treatment,the clinical efficacy of the two groups was evaluated,and the changes of Visual Analogue Scale(VAS)score and Japanese Orthopaedic Association(JOA)score were observed before and after treatment in the two groups.The changes of serum nuc lear factor-KB(NF-κB)p65,tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)were compared between the two groups before and after treatment.Results(1)After treatment,the JOA score and VAS score of the two groups were significantly improved(P＜0.05),and the improvement of JOA score and VAS score in the observation group was significantly superior to that in the control group,the differences were statistically significant(P＜0.05).(2)After treatment,the contents of NF-κB p65,serum IL-6 and TNF-α in the two groups were significantly improved(P＜0.05),and the improvement of NF-KB p65,serum IL-6 and TNF-α in the observation group was significantly superior to that in the control group,the differences were statistically significant(P＜0.05).(3)The total effective rate of the treatment group was 86.67％(26/30);the control group was 50.00％(15/30).The curative effect of the treatment group was significantly superior to that of the control group,and the difference was statistically significant(P＜0.05).Conclusion Lumbar fixed-point pulling method combined with magnetic needling in the treatment of LDH can effectively relieve the pain symptoms of patients,improve the lumbar function of patients,and effectively reduce the levels of serum NF-κB p65,IL-6 and TNF-α.The clinical effect is significant.

Objective:To investigate the role of levofloxacin combined with recombinant human granulocyte colony-stimulating factor(G-CSF)or only G-CSF supportive therapy in preventing infection in autologous hematopoietic stem cell transplantation(ASCT),and to analyze the length of hospital stay,hospitalization cost and post-transplant survival of the patients.Methods:A retrospective analysis was performed in the patients with hematological malignancies who accepted ASCT at our hospital from January 2012 to July 2022,the febrile neutropenia,the incidence of bacterial infection and the use rate of intravenous antibiotics in the levofloxacin+G-CSF group and only G-CSF support group during ASCT were observed.The length of hospital stay,total cost during hospitalization and survival after 90 days of transplantation between the two groups were compared.Results:A total of 102 cases were included in this study,including 57 cases of multiple myeloma,36 cases of acute leukaemia,7 cases of lymphoma,3 cases of myelodysplastic syndrome,1 case of light chain amyloidosis,and 1 case of POEMS syndrome.47 patients received levofloxacin+G-CSF antibacterial prophylaxis,and 55 patients received G-CSF supportive therapy.In the levofloxacin+G-CSF group,40 cases(85.11％)developed febrile neutropenia,and 13 cases(27.66％)were confirmed as bacterial infection.In the G-CSF group,44 cases(80.00％)developed febrile neutropenia,and 16 cases(29.09％)were bacterial infection.There was no statistically significant difference in the incidence of febrile neutropenia and bacterial infection between the two groups(x2=0.46,P=0.50;x2=0.03,P=0.87).The use rate of intravenous antibiotics in the levofloxacin+G-CSF group was 85.11％(40/47),which was not statistically different from 85.45％(47/55)in the G-CSF group(X2=0.04,P=0.84).The detection rates of levofloxacin-resistant bacteria in the levofloxacin+G-CSF group and G-CSF group were 8.57％(3/35)and 21.43％(6/28),respectively,with no statistical difference(x2=0.65,P＞0.05).The median length and median cost of hospitalization in the levofloxacin+G-CSF group and G-CSF group were 25 d vs 22 d and 78 216.24 yuan vs 80 724.38 yuan,with no statistically significant differences(t=3.00,P=0.09;t=0.94,P=0.09).Within 90 days after transplantation,two cases(4.26％)died in the levofloxacin+G-CSF group and one case(1.82％)died in the G-CSF group,with no statistically significant difference between the two groups(x2=0.53,P=0.47).Conclusion:Application of levofloxacin+G-CSF showed no significant benefit compared to G-CSF support for the prevention of bacterial infections during ASCT.

Objective:To study the molecular mechanism of functional defect of protein C (PC) caused by point mutations of human protein C gene (PROC) N355S,G392E and T314A. Methods:The wild-type and mutant plasmids (PCWT,PCN355S,PCG392E,PCT314A) of PROC gene were constructed and transiently transfected into HEK293 cells. The expression of mutant proteins in vitro were tested. The mRNA level changes of wild-type and mutant PC after 24 h of transfection were detected by real-time PCR. Western blot and ELISA were used to detect the changes of intracellular and extracellular protein levels of wild-type and mutant PC. The supernatant of cells transfected for 24-48 h was concentrated by ultrafiltration. The protein in the concentrated solution was quantified,and PC activation and enzyme kinetics tests were performed. Clustal Omega multiple sequence alignment was used to analyze the conservation of amino acid mutation sites. The effect of mutation on PC protein structure was analyzed by PyMOL software. Results:The relative expression abundances of PROC mRNA in PCN355S,PCG392E and PCT314A groups were 1.14±0.46,0.96±0.08 and 1.08±0.17,respectively,and there were no significant differences compared with 1.02±0.24 in PCWT group (P>0.05). Western blot analysis of the lysates of transfected cells showed that the content of PCT314A recombinant protein slightly decreased and the band became relatively lighter. The ELISA results of the concentrated cell culture supernatants showed that the PC:Ag levels of PCN355S and PCG392E were 98.8％±2.4％ and 101.4％±3.1％,respectively,with no significant differences compared with PCWT,while PCT314A decreased compared with PCWT (PC:Ag:88.6％±3.2％) (P<0.05). The results of enzyme kinetics test showed that APCN355S (Km=338.3±43.2,Vmax=2.015±0.12),APCG392E (Km=292.2±28.4,Vmax=1.893±0.07) and APCT314A (Km=299.5±24.6,Vmax=1.775±0.06) showed an increase in Km and a decrease in Vmax compared with APCWT (Km=238.2±4.58,Vmax=3.205±0.06). Multiple sequence alignment suggested that the three mutations be highly conservative in different species. The structural model suggested that the amino acid substitutions of N355S,G392E and T314A mutations collide with the surrounding amino acid groups,causing distortion of the surrounding structure,which may have adverse effects on the folding and biological function of PC. Conclusion:The N355S,G392E and T314A mutations in the PROC gene cause functional defects in PC by weakening the binding between PC and substrate. These three mutations have caused serious spatial collisions in the protein structure,affecting the folding of PC and the reactivity of active sites.

Objective:To assess the efficacy and toxicities of carboplatin+etoposide(CE)regimens combined with abiraterone+prednisone(AAP)in patients with metastatic castration-resistant prostate cancer(mCRPC)after progression with docetaxel+prednisone(DP)regimens chemotherapy and novel hormone therapy(NHT).Methods:Retrospective analysis of mCRPC treated with DP regimens chemotherapy and/or NHT after progression,received CE regimens with AAP every 3 weeks for one cycle×6 cycles.The outcome were prostate specific an-tigen(PSA)response rate,time to PSA progression(TTPP),radiographic progression-free survival(rPFS),30%reduction in PSA,90%reduc-tion in PSA,the objective response remission rate and overall survival(OS).Results:From March 2019 to February 2024,37 eligible mCRPC patients were admitted to Cancer Hospital of Huanxing Chaoyang District Beijing and National Cancer Center/National Cancer Clinical Re-search Center/Cancer Hospital.After progression,CE regimens combined with AAP regimens was used for treatment.The median follow-up was 12.0(3.0-57.0)months.The median treatment cycle was 4 cycles.The PSA response rate was 42.1%.The median TTPP was 4.0 months;the median rPFS was 8.9 months and the median OS was 15.0 months.The objective remission rate was 24.3%,the proportion of 30%de-crease in PSA was 59.5%,and the proportion of 90%decrease in PSA was 16.2%.As for treatment side effects,10 cases had grade 3 or higher adverse reactions.Conclusions:CE regimens combined with AAP for mCRPC patients who failed DP regimens chemotherapy and/or NHT initially showed good clinical efficacy and tolerability.Additional sample size and follow-up time are needed to further validate the effic-acy.

This study was aimed at exploring the effects of Toxoplasma gondii ROP16 Ⅰ/Ⅲ protein on the cell cycle,pro-liferation,and apoptosis of THP-1 cells via activation of STAT3 phosphorylation.ROP16 empty vector and recombinant lenti-viral overexpression vector were transfected into THP-1 cells,the fluorescence intensity was observed,and overexpression was verified.STAT3 and P-STAT3 protein expression in THP-1 cells overexpressing ROP16 Ⅰ/Ⅲ was detected by Western blot-ting.The cells were treated with the STAT3 phosphorylation inhibitor Stattic,and divided into a THP-1 group,THP-1+Stattic group,THP-1-ROP16 Ⅰ/Ⅲ group,and THP-1-ROP16Ⅰ/Ⅲ+Stattic group.Changes in the cell cycle,proliferation curve,and apoptosis rate were detected with flow cytometry and CCK-8 assays,and the expression levels of the apoptosis-related proteins Bax,Cleaved Caspase3,Caspase9,and Bcl-2 were detected by Western blotting.A strong fluorescence signal was ob-served after transfection of ROP16 recombinant lentivirus overexpression vector into THP-1 cells,and ROP16 mRNA and pro-tein expression significantly increased(FmRNA=314.1,Fprotein=758.7,P＜0.001),thus indicating successful construction of cell strains with stable overexpression.In addition,P-STAT3 protein showed higher expression in THP-1-ROP16 Ⅰ/Ⅲ group than the THP-1 group(F=606.1,P＜0.001).Flow cytometry and CCK-8 results indicated that,compared with the THP-1 group,the THP-1-ROP16 Ⅰ/Ⅲ group showed a cell cycle blocked in G1 phase,lower cell proliferation ability(t Ⅰgroup=19.2,tⅢgroup=24.0,P＜0.01),and a higher apoptosis rate(t Ⅰgroup=175.1,tⅢgroup=205.2,P＜0.001).Western blotting results also demonstrated significantly elevated expression of the intracellular pro-apoptotic proteins Bax,Cleaved Caspase3,and Caspase9(t Bax Ⅰgroup=15.8,tBaxⅢgroup=11.1,t Caspase9 Ⅰ group=9.1,t Caspaseg Ⅲ group=10.2,t Cleaved Caspase3 Ⅰ group=16.5,t Cleaved Caspase3 Ⅲ group=12.9,all P＜0.001),and significantly diminished expression of the anti-apoptotic protein Bcl-2(t Ⅰgroup=18.4,tⅢgroup=26.2,P＜0.001).However,after Stattic treatment,the cell cycle,proliferation ability,apoptosis rate,and expression of apoptosis-re-lated proteins in the THP-1-ROP16 Ⅰ/Ⅲ+Stattic group were restored to near baseline levels.Thus,Toxoplasma gondii ROP16 Ⅰ/Ⅲ proteins were found to block the THP-1 cell cycle at G1 phase,inhibit cell proliferation,and promote apoptosis by activating the phosphorylation of STAT3.

This study was aimed at exploring the effects of Toxoplasma gondii ROP16 Ⅰ/Ⅲ protein on the cell cycle,pro-liferation,and apoptosis of THP-1 cells via activation of STAT3 phosphorylation.ROP16 empty vector and recombinant lenti-viral overexpression vector were transfected into THP-1 cells,the fluorescence intensity was observed,and overexpression was verified.STAT3 and P-STAT3 protein expression in THP-1 cells overexpressing ROP16 Ⅰ/Ⅲ was detected by Western blot-ting.The cells were treated with the STAT3 phosphorylation inhibitor Stattic,and divided into a THP-1 group,THP-1+Stattic group,THP-1-ROP16 Ⅰ/Ⅲ group,and THP-1-ROP16Ⅰ/Ⅲ+Stattic group.Changes in the cell cycle,proliferation curve,and apoptosis rate were detected with flow cytometry and CCK-8 assays,and the expression levels of the apoptosis-related proteins Bax,Cleaved Caspase3,Caspase9,and Bcl-2 were detected by Western blotting.A strong fluorescence signal was ob-served after transfection of ROP16 recombinant lentivirus overexpression vector into THP-1 cells,and ROP16 mRNA and pro-tein expression significantly increased(FmRNA=314.1,Fprotein=758.7,P＜0.001),thus indicating successful construction of cell strains with stable overexpression.In addition,P-STAT3 protein showed higher expression in THP-1-ROP16 Ⅰ/Ⅲ group than the THP-1 group(F=606.1,P＜0.001).Flow cytometry and CCK-8 results indicated that,compared with the THP-1 group,the THP-1-ROP16 Ⅰ/Ⅲ group showed a cell cycle blocked in G1 phase,lower cell proliferation ability(t Ⅰgroup=19.2,tⅢgroup=24.0,P＜0.01),and a higher apoptosis rate(t Ⅰgroup=175.1,tⅢgroup=205.2,P＜0.001).Western blotting results also demonstrated significantly elevated expression of the intracellular pro-apoptotic proteins Bax,Cleaved Caspase3,and Caspase9(t Bax Ⅰgroup=15.8,tBaxⅢgroup=11.1,t Caspase9 Ⅰ group=9.1,t Caspaseg Ⅲ group=10.2,t Cleaved Caspase3 Ⅰ group=16.5,t Cleaved Caspase3 Ⅲ group=12.9,all P＜0.001),and significantly diminished expression of the anti-apoptotic protein Bcl-2(t Ⅰgroup=18.4,tⅢgroup=26.2,P＜0.001).However,after Stattic treatment,the cell cycle,proliferation ability,apoptosis rate,and expression of apoptosis-re-lated proteins in the THP-1-ROP16 Ⅰ/Ⅲ+Stattic group were restored to near baseline levels.Thus,Toxoplasma gondii ROP16 Ⅰ/Ⅲ proteins were found to block the THP-1 cell cycle at G1 phase,inhibit cell proliferation,and promote apoptosis by activating the phosphorylation of STAT3.

OBJECTIVE: To explore the clinical characteristics of hospitalized patients with hematologic diseases complicated with carbapenem-resistant organisms (CRO) infection and analyze the risk factors of 30-day all-cause mortality. METHODS: The clinical data and laboratory test data of 77 hospitalized patients with hematologic diseases complicated with CRO infection in department of hematology of the Third Hospital of Shanxi Medical University from January 2015 to December 2020 were retrospectively analysed, the risk factors of 30-day all-cause mortality after CRO infection were analyzed by multivariate logistic regression. RESULTS: Among the total of 77 patients with hematologic diseases complicated with CRO infection, 29 died and 48 survived within 30 days of infection, with a case fatality rate of 37.66%. A total of 93 strains of CRO were isolated from these patients, of which Acinetobacter baumannii had the highest detection rate (25.81%, 24/93), followed by Pseudomonas aeruginosa (18.28%, 17/93). The lung was the most common site of CRO infection. The detected pathogens were highly resistant to carbapenems, and 64.52% (60/93) of the pathogens were resistant to imipenem with minimum inhibitory concentration (MIC)≥16 μg/ml. The results of the univariate analysis showed that albumin concentration <25 g/L (P =0.048), serum creatinine concentration≥120 μmol/L (P =0.023), age-adjusted Charlson comorbidity index (ACCI) (P =0.037) and primary treatments (supportive treatment, immunosuppressive therapy, chemotherapy, HSCT) (P =0.048) were significantly associated with 30-day all-cause mortality after infection. The results of multivariate logistic regression analysis showed that when CRO infection confirmed, albumin concentration <25 g/L (P =0.014, OR=6.171), serum creatinine concentration≥120 μmol/L (P =0.009, OR=10.867) were independent risk factors for 30-day mortality of patients with hematologic diseases complicated with CRO infection. CONCLUSION The mortality rate of CRO-infected patients with hematologic diseases is high. The detected pathogenic bacteria are highly resistant to imipenem. The albumin concentration <25 g/L and the serum creatinine concentration≥ 120 μmol/L at diagnosis of CRO infection were independent risk factors for 30-day mortality of the patients with hematologic diseases.
Humans ; Carbapenems/pharmacology* ; Retrospective Studies ; Creatinine ; Hematologic Diseases ; Risk Factors ; Imipenem ; Albumins

Humans ; Philadelphia Chromosome ; Neoplasms ; Myelodysplastic Syndromes/genetics* ; Disease Progression

ObjectiveTo predict the potential targets and mechanism of Jingfang mixture in the treatment of H1N1 influenza and provide references for clinical application of Jingfang mixture. MethodThe active components and targets of Jingfang mixture against H1N1 influenza were screened out by Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform（TCMSP），SwissTargetPrediction， and TargetNet. The targets of H1N1 influenza were obtained from GeneCards，Online Mendelian Inheritance in Man （OMIM）， and DisGeNET and standardized by UniProt KB. The intersection targets were obtained by Venny 2.1.0. The "drug-component-target" network was constructed with Cytoscape 3.2.1 and analyzed for the topological attributes. The intersection targets were uploaded to STRING 11.5 to obtain the protein-protein interaction （PPI） network. Gene Ontology （GO） enrichment analysis and Kyoto Encyclopedia of Genes and Genomes （KEGG） analysis were carried out by Metascape. Finally，the top active components ranked by degree were docked to the core targets by Autodock vina and visually analyzed by PyMOL. Balb/c female rats were used for experimental verification. Hematoxylin-eosin（HE） staining was used to observe the pathological changes in lung tissues. Enzyme-linked immunosorbent assay（ELISA）was used to detect the levels of tumor necrosis factor-α（TNF-α），interleukin-10（IL-10）， and interleukin-17（IL-17）. Real-time fluorescence-based quantitative polymerase chain reaction（Real-time PCR） and Western blot were used to detect the mRNA and protein expression levels in lung tissues. ResultThere were 144 active components in Jingfang mixture. A total of 421 target genes of Jingfang mixture and 2 956 targets of H1N1 influenza were identified，including 199 common targets. Topological analysis showed that the core components of Jingfang mixture against H1N1 influenza included quercetin，luteolin， and kaempferol，and the core targets included prostaglandin-endoperoxide synthase 2（PTGS2），estrogen receptor alpha（ESR1），inducible nitric oxide synthase 2（iNOS2），peroxisome proliferator-activated receptorγ（PPARγ），and cyclooxygenase-1（PTGS1）. GO enrichment yielded 697 items in biological process （BP） （P<0.01）， 59 items in molecular function （MF）（P<0.01）， and 21 items in cellular component （CC） （P<0.01）. A total of 132 signaling pathways （P<0.01） were obtained by KEGG enrichment analysis， including phosphatidylinositol 3-kinases（PI3K）/protein kinase B（Akt） signaling pathway and mitogen-activated protein kinase（MAPK） signaling pathway，most of which were related to the regulation of immune inflammation. Molecular docking showed that the binding energy of the active components of Jingfang mixture to the core targets was less than -5.0 kcal·mol^-1，indicating good binding activity. HE staining showed that the lung tissues were significantly improved after drug intervention，and Real-time PCR and Western blot showed that Jingfang mixture could reduce the mRNA and protein expression of PI3K and Akt in lung tissues. ConclusionJingfang mixture can play an anti-viral effect against the influenza A virus through multiple components，multiple targets， and multiple pathways. The active components quercetin，luteolin， and kaempferol may control the inflammation and regulate immunity on the PI3K/Akt，MAPK， and other signaling pathways by acting on targets such as PTGS2，ESR1，iNOS2，PPARγ， and PTGS1.