AIM:To investigate the inhibitory ef-fect of Zushima ointment on inflammation and bone destruction in CIA mice.METHODS:SPF grade male DBA/1 mice were used,6 were random-ly selected as the normal group,and 18 CIA mice that were successfully modelled were randomly di-vided into the model group,the plaster group(1.0 g/kg),and the fuselage group(0.12 g per time)ac-cording to the random number table method,6 mice in each group,and each administered group was given medication according to the body mass,and saline was given to both the normal and model groups.The normal group and the model group were given saline,and breathable adhesive paper was applied once a day for 4 h/session for 4 consec-utive weeks.The arthritis scoring index was used to observe the changes of arthritis symptoms and ar-thritis index scores of mice in each group.Micro-CT was used to observe the damage of hind paw of mice,real-time fluorescence PCR was used to de-tect the mRNA expression of IL-17,IL-1β and TNF-αin ankle joint tissues,and immunohistochemistry was used to detect the expression of OPG and RANKL proteins in ankle joint tissues,and hematox-ylin-eosin(HE)staining was used to observe the pathological changes of synovial tissues after the treatment.The pathological changes of synovial tis-sue were observed after hematoxylin-eosin(HE)staining,and the changes of osteoclasts in ankle joint tissue were observed by anti-tartaric acid phosphatase(TRAP)method.RESULTS:Compared with the normal group,the arthritis index score of the model mice was significantly higher(P＜0.05).Micro-CT showed severe bone erosion in the hind paws of the mice,destruction of the bone surface and reduction of bone volume.The expression of IL-17,IL-1β and TNF-α mRNA(PCR)in the ankle joint tissues was significantly higher(P＜0.05).Im-munohistochemistry showed that the relative ex-pression of OPG protein in the ankle joint tissues was reduced(P＜0.05).Immunohistochemistry showed a decrease in the relative expression of OPG protein(P＜0.01)and an increase in the rela-tive expression of RANKL protein(P＜0.01).HE re-sults showed moderate inflammatory cell infiltra-tion,swelling of synovial cells,massive formation of vascular opacities and synovial hyperplasia;an increase in the number of osteoclasts,roughness of the surface of articular cartilage tissue,severe bone destruction and thinning of the cartilage lay-er.Compared with the model group,the arthritic symptoms of mice in the cream group and the futa-lin group were relieved and the arthritis index score was reduced;the bone density of the mice's hind paws improved,effectively relieving osteopo-rosis;the expression of IL-17,IL-1β and TNF-αmRNA(PCR)in the ankle joint tissue was signifi-cantly reduced(P＜0.05);the immunohistochemical results showed that the relative expression of OPG protein was increased(P＜0.05),the relative expres-sion of RANKL protein decreased(P＜0.01).HE re-sults showed that synovial cell enlargement was significantly improved,mild inflammatory cell infil-tration,synovial hyperplasia was not obvious;the number of broken bone was reduced,articular car-tilage destruction was significantly improved and relieved,and the thickness of cartilage layer was significantly increased.CONCLUSION:Ancestral hemp poultice relieves local symptoms of RA,re-duces the expression of inflammatory factors and attenuates the inflammatory response,possibly by inhibiting osteoclast differentiation and activation through modulation of the OPG/RANKL signalling axis,which further ameliorates the biological ef-fects of articular bone and cartilage destruction.

Objective:To investigate the effect of copper chlorophyllin sodium salt(CHL)on the sensitivity of human pancreatic cancer cells in response to gemcitabine(GEM)therapy and on the therapeutic effect on pancreatic cancer cells that have developed GEM resistance.Methods:MIA GR(a pancreatic cancer cell line resistant to GEM)was induced by a low-dose continuous incremental method,and the half inhibitory concentration(IC50)of MIA WT and MIA GR to GEM treatment was detected by the CCK-8 method,and the resistance index was calculated;the difference in IC50 of CHL on the two types of cells was detected by the CCK-8 method after treating MIA WT and MIA GR cells with different concentrations of CHL,CCK-8 method was used to detect the difference in IC50 of CHL on the two types of cells;on the basis of IC50,MIA WT and MIA GR cells were intervened with CHL and(or)GEM with different multiplicity of IC50,respectively,and the growth inhibition curves of MIA WT and MIA GR cells were detected by the CCK-8 method under the intervention of CHL combined with GEM;After the intervention of MIA WT and MIA GR cells with CHL and(or)GEM at IC50,respectively,the effects on the proliferation of the two different cells were detected using the clone formation assay;the effects on cytotoxicity/activity were observed under fluorescence microscopy;and the effects on apoptosis were detected using flow cytometry.Finally,western blotting was used to detect the effects of CHL and(or)GEM interventions on the drug resistance-associated molecules P-glycoprotein(P-gp)and ribonucleotide reductase regulatory subunit M2(RRM2)in MIA GR cells,the and sensitivity-related molecule deoxycytidine kinase(DCK)on protein expression levels.Results:MIA GR cells were verified to be well drug resistant,with resistance indices of 549.1 and 667.9 after 48 h and 96 h after GEM intervention compared to homologous wild-type MIA WT cells,respectively;CHL intervention inhibited the proliferation of MIA GR cells more significantly compared to that of MIA WT cells;and CHL in combination with GEM exerted a more significant growth inhibitory effect compared to GEM alone in both MIA WT cells(P<0.001)and MIA GR cells(P<0.01).CHL significantly inhibited the tumor proliferation of MIA GR cells,and the inhibitory effect was more pronounced in both cells when combined with GEM(P<0.000 1);furthermore,compared to GEM alone,the intervention with CHL could cause more pronounced cytotoxicity(P<0.000 1)in both MIA WT and MIA GR cells.caused more pronounced cytotoxicity(P<0.000 1)and induced a higher percentage of apoptosis than GEM alone.The results of the western blotting assay showed that CHL intervention caused a decrease in the expression levels of P-gp and RRM2 proteins,as well as an increase in the protein expression level of DCK in MIA GR cells.Conclusion:CHL increases the sensitivity of pancreatic cancer cells to GEM and also induces a decrease in the resistance of drug-resistant pancreatic cancer cells to GEM.

Background and purpose:Pancreatic cancer is a highly aggressive solid tumor of the digestive system,with radical resection being unfeasible in approximately 80%of patients due to the absence of specific clinical manifestations in the early stages.The use of gemcitabine as a first-line chemotherapeutic agent has not significantly improved patient prognosis,primarily due to the development of chemoresistance.The precise mechanisms underlying gemcitabine resistance in pancreatic cancer remain unclear.This study aimed to explore potential biomarkers associated with gemcitabine chemoresistance in pancreatic cancer by utilizing gemcitabine-resistant cell lines and pathological pancreatic cancer tissues,in conjunction with data from online databases.Additionally,we analyzed follow-up data from pancreatic cancer patients to assess the impact of relevant targets on patient prognosis.Methods:In this study,gemcitabine-resistant cell lines were developed through stepwise induction using a gemcitabine concentration gradient.Second-generation high-throughput RNA-seq sequencing was conducted on these resistant cells,and bioinformatics analysis was employed to identify four pancreatic cancer genes from the Gene Expression Omnibus(GEO)datasets(GSE106336,GSE110580,GSE35141,and GSE140077).Co-expressed genes were screened using real-time fluorescence quantitative polymerase chain reaction(RTFQ-PCR),Western blot and immunohistochemistry to verify the expression levels of target molecules.Surgical specimens from 70 patients diagnosed with pancreatic adenocarcinoma at the First Affiliated Hospital of Xi'an Jiao Tong University between June 2018 and June 2021 were analyzed.These included 30 specimens resistant to gemcitabine,16 non-resistant specimens,and 24 normal pancreatic tissues as controls.Ethical approval was obtained(Ethical approval:2021 LunxuanKeZi No.457,No.XJTU1AF2021LSK-457).Clinical and prognostic information was collected,and the log-rank test was used to evaluate the relationship between target molecule expression and patient prognosis.Results:The half maximal inhibitory concentration(IC50)for gemcitabine was significantly higher in the gemcitabine-resistant cell strain(Mia GR)than in the wild-type cell line(Mia WT)(258.10 μmol/L vs 0.18 μmol/L),with a resistance index(RI)of 1 443.9.Transcriptome sequencing identified 3 985 differentially expressed genes,of which 25 were shared with the GEO datasets.Further analysis highlighted INPP4B as a key gene.RTFQ-PCR and Western blot confirmed that INPP4B mRNA and protein levels were significantly elevated in drug resistant cells compared to wild-type cells(P＜0.05).Immunohistochemical analysis revealed that INPP4B expression was significantly higher in drug resistant pancreatic cancer tissues compared to non-drug resistant tissues,and lower in normal tissues than in both cancerous tissue types.Kaplan-Meier curves demonstrated that patients with low INPP4B expression had significantly better progression-free survival(PFS)than those with high expression(HR=2.874,95%CI:1.262-6.544,P=0.013).Although patients with low INPP4B expression also showed better overall survival(OS),the difference was not statistically significant(HR=1.484,95%CI:0.518-4.250,P=0.465).Conclusion:INPP4B may serve as a potential biomarker for gemcitabine chemoresistance in pancreatic cancer and is associated with poor prognosis in drug resistant patients.Developing targeted assays and treatments for INPP4B could facilitate early identification of patients likely to exhibit resistance to gemcitabine therapy,thereby improving their prognosis.

Background and purpose:Pancreatic cancer is a highly aggressive solid tumor of the digestive system,with radical resection being unfeasible in approximately 80%of patients due to the absence of specific clinical manifestations in the early stages.The use of gemcitabine as a first-line chemotherapeutic agent has not significantly improved patient prognosis,primarily due to the development of chemoresistance.The precise mechanisms underlying gemcitabine resistance in pancreatic cancer remain unclear.This study aimed to explore potential biomarkers associated with gemcitabine chemoresistance in pancreatic cancer by utilizing gemcitabine-resistant cell lines and pathological pancreatic cancer tissues,in conjunction with data from online databases.Additionally,we analyzed follow-up data from pancreatic cancer patients to assess the impact of relevant targets on patient prognosis.Methods:In this study,gemcitabine-resistant cell lines were developed through stepwise induction using a gemcitabine concentration gradient.Second-generation high-throughput RNA-seq sequencing was conducted on these resistant cells,and bioinformatics analysis was employed to identify four pancreatic cancer genes from the Gene Expression Omnibus(GEO)datasets(GSE106336,GSE110580,GSE35141,and GSE140077).Co-expressed genes were screened using real-time fluorescence quantitative polymerase chain reaction(RTFQ-PCR),Western blot and immunohistochemistry to verify the expression levels of target molecules.Surgical specimens from 70 patients diagnosed with pancreatic adenocarcinoma at the First Affiliated Hospital of Xi'an Jiao Tong University between June 2018 and June 2021 were analyzed.These included 30 specimens resistant to gemcitabine,16 non-resistant specimens,and 24 normal pancreatic tissues as controls.Ethical approval was obtained(Ethical approval:2021 LunxuanKeZi No.457,No.XJTU1AF2021LSK-457).Clinical and prognostic information was collected,and the log-rank test was used to evaluate the relationship between target molecule expression and patient prognosis.Results:The half maximal inhibitory concentration(IC50)for gemcitabine was significantly higher in the gemcitabine-resistant cell strain(Mia GR)than in the wild-type cell line(Mia WT)(258.10 μmol/L vs 0.18 μmol/L),with a resistance index(RI)of 1 443.9.Transcriptome sequencing identified 3 985 differentially expressed genes,of which 25 were shared with the GEO datasets.Further analysis highlighted INPP4B as a key gene.RTFQ-PCR and Western blot confirmed that INPP4B mRNA and protein levels were significantly elevated in drug resistant cells compared to wild-type cells(P＜0.05).Immunohistochemical analysis revealed that INPP4B expression was significantly higher in drug resistant pancreatic cancer tissues compared to non-drug resistant tissues,and lower in normal tissues than in both cancerous tissue types.Kaplan-Meier curves demonstrated that patients with low INPP4B expression had significantly better progression-free survival(PFS)than those with high expression(HR=2.874,95%CI:1.262-6.544,P=0.013).Although patients with low INPP4B expression also showed better overall survival(OS),the difference was not statistically significant(HR=1.484,95%CI:0.518-4.250,P=0.465).Conclusion:INPP4B may serve as a potential biomarker for gemcitabine chemoresistance in pancreatic cancer and is associated with poor prognosis in drug resistant patients.Developing targeted assays and treatments for INPP4B could facilitate early identification of patients likely to exhibit resistance to gemcitabine therapy,thereby improving their prognosis.

Objective:To investigate the effect of copper chlorophyllin sodium salt(CHL)on the sensitivity of human pancreatic cancer cells in response to gemcitabine(GEM)therapy and on the therapeutic effect on pancreatic cancer cells that have developed GEM resistance.Methods:MIA GR(a pancreatic cancer cell line resistant to GEM)was induced by a low-dose continuous incremental method,and the half inhibitory concentration(IC50)of MIA WT and MIA GR to GEM treatment was detected by the CCK-8 method,and the resistance index was calculated;the difference in IC50 of CHL on the two types of cells was detected by the CCK-8 method after treating MIA WT and MIA GR cells with different concentrations of CHL,CCK-8 method was used to detect the difference in IC50 of CHL on the two types of cells;on the basis of IC50,MIA WT and MIA GR cells were intervened with CHL and(or)GEM with different multiplicity of IC50,respectively,and the growth inhibition curves of MIA WT and MIA GR cells were detected by the CCK-8 method under the intervention of CHL combined with GEM;After the intervention of MIA WT and MIA GR cells with CHL and(or)GEM at IC50,respectively,the effects on the proliferation of the two different cells were detected using the clone formation assay;the effects on cytotoxicity/activity were observed under fluorescence microscopy;and the effects on apoptosis were detected using flow cytometry.Finally,western blotting was used to detect the effects of CHL and(or)GEM interventions on the drug resistance-associated molecules P-glycoprotein(P-gp)and ribonucleotide reductase regulatory subunit M2(RRM2)in MIA GR cells,the and sensitivity-related molecule deoxycytidine kinase(DCK)on protein expression levels.Results:MIA GR cells were verified to be well drug resistant,with resistance indices of 549.1 and 667.9 after 48 h and 96 h after GEM intervention compared to homologous wild-type MIA WT cells,respectively;CHL intervention inhibited the proliferation of MIA GR cells more significantly compared to that of MIA WT cells;and CHL in combination with GEM exerted a more significant growth inhibitory effect compared to GEM alone in both MIA WT cells(P<0.001)and MIA GR cells(P<0.01).CHL significantly inhibited the tumor proliferation of MIA GR cells,and the inhibitory effect was more pronounced in both cells when combined with GEM(P<0.000 1);furthermore,compared to GEM alone,the intervention with CHL could cause more pronounced cytotoxicity(P<0.000 1)in both MIA WT and MIA GR cells.caused more pronounced cytotoxicity(P<0.000 1)and induced a higher percentage of apoptosis than GEM alone.The results of the western blotting assay showed that CHL intervention caused a decrease in the expression levels of P-gp and RRM2 proteins,as well as an increase in the protein expression level of DCK in MIA GR cells.Conclusion:CHL increases the sensitivity of pancreatic cancer cells to GEM and also induces a decrease in the resistance of drug-resistant pancreatic cancer cells to GEM.

Objective:To compare the effects of ultrasound-guided dynamic needle tip positioning (DNTP) and long axis in-plane (LAX-IP) techniques for axillary vein puncture and catheterization.Methods:One hundred Society of Anesthesiologists physical statusⅠ-Ⅲ patients of both sexes, aged 18-64 yr, with body mass index of 20-28 kg/m 2, scheduled for elective axillary vein cannulation, were divided into 2 groups ( n=49 each) using the random number table method: DNTP group and LAX-IP group.Axillary vein puncture was performed using DNTP technique and LAX-IP technique under ultrasound guidance in DNTP group and LAX-IP group, respectively.Successful puncture at first attempt, overall successful catheterization, the number of needle tip redirection, and axillary vein puncture time and catheterization time were recorded.The occurrence of complications such as axillary artery puncture, posterior wall penetration of axillary vein, hematoma formation, pneumothorax, and nerve injury was recorded. Results:Compared with group LAX-IP, the success rate of puncture at first attempt was significantly increased, the number of cases required needle redirection was decreased, and the puncture time was shortened ( P<0.05), and no significant change was found in the logarithm of the posterior wall penetration of axillary vein in group DNTP ( P>0.05). No complications such as arterial puncture, hematoma, pneumothorax, or nerve injury occurred in two groups. Conclusions:Compared with LAX-IP technique, ultrasound-guided DNTP technique can dynamically observe the position of the needle tip, the operation is simple and safe, and it is worthy of clinical promotion when used for axillary vein puncture and cannulation.

Objective:To investigate the clinical curative effect of closed reduction and internal fixation of Jakob type II lateral humeral condyle fractures in children under ultrasound guidance.Methods:A retrospective case series analysis was made on clinical data of 59 patients with Jakob type II lateral humeral condyle fractures treated at Children's Hospital of Chongqing Medical University from August 2016 to August 2017. There were 30 males and 29 females, with the age of 1.5-8.1 years [(4.0±1.8)years]. There were 34 patients treated by open reduction and internal fixation and monitored by the X-ray (control group), and 25 patients treated by closed reduction and internal fixation and monitored by ultrasound (study group). The operation time, bleeding volume, fracture healing time, and incidence of complications were compared between the two groups. The elbow joint function was evaluated by Broberg and Morrey standard at the latest follow-up.Results:All patients were followed up for 17-31 months[(23.2±4.2)months]. The operation time and bleeding volume in control group were (50.7±22.2)minutes and (6.1±3.8)ml, obviously higher than those in study group [(21.4±3.3)minutes, (1.1±0.3)ml] ( P<0.05). The fracture healing time was (8.0±0.8)weeks in control group and (7.8±0.7)weeks in study group ( P>0.05). According to the Broberg and Morrey standard, the good and excellent rate of elbow joint function in control group was 97%, with excellent results in 31 patients, good in 2, fair in 1, and poor in 0; the good and excellent rate of elbow joint function in study group was 100%, with excellent results in 22 patients, good in 3, fair in 0 and poor in 0 ( P>0.05). In study group, wound infection or malunion was not seen, and only two patients showed postoperative wire tail irritability and recovered after the removal of wires. While in control group, wound infection was seen in 3 patients and malunion was observed in 2 patients, but all patients were with distal humerus lateral bone formations. The incidence of complications was 15% in control group, higher than 0% in study group ( P<0.05). Conclusion:Compared to open reduction internal fixation, ultrasound-guided closed reduction and internal fixation of Jakob type Ⅱ lateral humeral condyle fractures in children has similar therapeutic effect, but it can shorten operation time and reduce bleeding and complications.

Objective:To evaluate the analgesic efficacy of pericapsular nerve group (PENG) block in elderly patients undergoing hip replacement under subarachnoid block.Methods:Fifty patients of both sexes, aged 65-89 yr, of American Society of Anesthesiologists physical status Ⅱ or Ⅲ, with body mass index 20-30 kg/m 2, undergoing unilateral hip arthroplasty, were divided into 2 groups ( n=25 each) by a random number table method: PENG group and fascia iliaca compartment block (FICB) group.In PENG group, 0.4% ropivacaine hydrochloride 20 ml was injected around the nerve innervating the hip joint capsule under ultrasound guidance.In FICB group, 0.4% ropivacaine 30 ml was injected around the nerve innervating the fascia iliaca compartment under ultrasound guidance.Subarachnoid block was performed in both groups.Visual analog scale scores and scores for satisfaction with analgesia at rest and during activity were recorded before blockade (T 0), at 10, 20 and 30 min after blockade (T 1-3) and when placed in the position for spinal anesthesia (T 4). The cumulative consumption of sufentanil, effective pressing times of analgesic pump, and development of related complications were recorded at 6, 12, 24 and 48 h after operation (T 5-8). Results:Compared with FICB group, the VAS scores at rest and during activity were significantly decreased at T 1-4, and scores for satisfaction with analgesia during activity were increased in PENG group ( P<0.05). There was no significant difference between the two groups in the cumulative consumption of sufentanil and effective pressing times of analgesic pump ( P>0.05). One patient developed postoperative delirium in group FICB, and no patients developed puncture site infection and nerve damage after operation in two groups. Conclusion:PENG block produces better analgesic efficacy than FICB when used for elderly patients undergoing hip replacement under subarachnoid block.

A novel method for accurate,fast and sensitive detection of pesticides such as imidacloprid,isocarbophos,phoxim,dursban,imidacloprid,pyridaben and avermectin in environmental water samples has been developed by using ultrasound-assisted dispersive liquid-liquid microextraction (UA-DLLME) coupled with high performance liquid chromatography with ultraviolet detection (HPLC-UV).The UA-DLLME parameters such as types/volumes of extraction/dispersion solvents,ultrasonic time,ionic strength and extraction time were investigated.Under the optimized extraction conditions,the linearity for the detection of six pesticides in the concentration range of 10-600 μg/L was obtained with limits of detections (LODs) of 0.8-3.1 μg/L and relative standard deviations (RSDs) of 4.7％-11.3％.UA-DLLME method exhibited strong enrichment ability for the six pesticides,and the enrichment factor (EFs) were ranged from 58 to 187.This method had perfect linearity,precision and recovery results,and showed obvious advantages and practicality comparing the previously reported methods.

Objective To observe the effect of tendon-bone healing after anterior cruciate ligament reconstruction through BFGF and PDGF.Methods Based of seventy two healthy matured New Zealand white rabbits underwent ACL reconstruction.Devide three groups through different methods,group BFGF,group PDGF,group normal.Results The maximum loads of group BFGF and PDGF after 2 and 5 weeks are higher than group normal,there was significant difference in maximum loads (P <0.05).The max stiffness of group BFGF and PDGF after 2 and 5 weeks are higher than group normal,there was significant difference in maximum loads(P <0.05).Conclusion The group BFGF and PDGF can promote tendon-bone healing by increasing the vascularization and blood vessels of the tendon-bone interface and vascular endothelial growth factor.In biomechanics,group BFGF and PDGF can pro-mote maximum loads、max stiffness and hardness of tendon-bone healing.