Objective To investigate short-term clinical efficacy of autologous leukocyte-poor platelet-rich plasma(LP-PRP)treatment of knee osteoarthritis(KO A).Methods 85 cases of patients with Keligren Lawrence grade Ⅰ-Ⅲ knee os-teoarthritis in Peking University First Hospital Taiyuan Hospital(Taiyuan Central Hospital)from 2022 to 2023 were collect-ed for autologous LP-PRP collection and quality assessment using a blood component separator,and all patients were treated with autologous LP-PRP.The degree and function of knee pain were assessed by visual analog scale(VAS)and knee arthri-tis index scale(WOMAC)at 1,3 and 6 months after injection.Knee MRI was performed after 6 months of treatment,and the MRI imaging changes before and after treatment were compared.Different influencing factors in the treatment results were grouped and analyzed,mainly including platelet concentration in LP-PRP and K-L grading of knee joint.According to the platelet concentration in LP-PRP,it was divided into three grades,which are low concentration[(＜800)×109/L],medium concentration[(800-1 000)×109/L],and high concentration[(＞1 000)× 109/L];According to the K-L grade of the knee joint,the severity of knee osteoarthritis was divided into three grades:Ⅰ、Ⅱ、Ⅲ.Results The VAS and WOMAC scores at 1,3 and 6 months after LP-PRP treatment were significantly lower than those before treatment,and the difference was sta-tistically significant(P＜0.05).There was a statistically significant difference in the therapeutic effect of different levels of platelet concentration,and when the platelet concentration was more than 1 000×109/L,the significant effect was the most obvious(P＜0.05).The therapeutic effect of different levels of platelet concentration was statistically significant(P＜0.05).MRI showed that the articular cartilage signal was significantly improved after treatment.Conclusion Autologous LP-PRP injection into knee cavity for the treatment of KO A has a good short-term clinical effect in relieving knee pain.

Animals ; Coronary Vessels ; Diabetes Mellitus ; Muscle, Smooth, Vascular ; Myocytes, Smooth Muscle ; Rats

Objective:To investigate the status of transition shock, career adaptation and professional identity among nursing interns, and to explore the mediating effect of professional identity on career adaptation and transition shock.Methods:In February 2022, 500 nursing students from 18 nursing colleges in Henan province and Shaanxi province were selected as the research subjects by convenient sampling method. General data questionnaire, Professional Identity Questionnaire of Nursing Students, Career Adaptability Scale of college students and Chinese version of Transition Impact Assessment Scale of Nursing Undergraduate Interns were used to conduct the questionnaire survey.Results:The career adaptability score of nursing intern was (133.59 ± 22.58); professional identity score was (63.35 ± 14.06); transition impact score was (45.18 ± 10.17). Transition shock was negatively correlated with career adaptation and professional identity ( r=-0.328, -0.484, both P<0.01). Occupational identity had a partial mediating effect on career adaptation and transition shock, accounting for 64.29% of the total effect. Conclusions:The transition shock of nursing interns is at medium level. Nursing educators can reduce the degree of transition shock by improving their professional identity to nursing and their own career adaptation, so as to better adapt to the professional role.

Objective:To evaluate the role of ferroptosis in the dorsal root gangions in neuropathic pain (NP) in rats.Methods:Thirty-two healthy male Sprague-Dawley rats, aged 6-8 weeks, weighing 180-220 g, were randomized into 4 groups ( n=8 each) using a random number table method: sham operation group (Sham group), NP group, NP+ solvent control group (NP+ Veh group), and NP+ liproxstatin-1 (Lip-1) group (NP+ Lip group). NP was induced by chronic constrictive injury (CCI) to sciatic nerve in anesthetized animals.In NP+ Lip group, liproxstatin-1 (diluted to 10 μg/μl in DMSO) 30 μl was intrathecally injected for 3 consecutive days after surgery.NP+ Veh group received intrathecal injection of DMSO 30 μl for 3 consecutive days after surgery.Mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) were measured at 3 days before surgery and on days 1, 3, 5, 7 and 10 after surgery.Rats were sacrificed after the end of pain threshold measurement on day 10 after surgery, and DRGs of the lumbar segment (L 3-5) on the left side were removed for determination of the levels of iron ion, reactive oxygen species (ROS), malondialdehyde (MDA), glutathione peroxidase (GSH-Px), and superoxide dismutase (SOD), expression of glutathione peroxidase 4 (GPX4) and acyl-CoA synthetase long-chain family member 4 (ACSL4) (by Western blot), and expression of ACSL4 in each nerve cells of DRGs (by immunofluorescence) and for microscopic examination of the ultrastructure of mitochondria in DRGs (by transmission electron microscopy). Results:Compared with Sham group, MWT was significantly decreased and TWL was shortened at T 2-6, levels of iron ions, ROS and MDA in DGRs were increased, activities of SOD and GSH-Px were decreased, ACSL4 expression was up-regulated, GPX4 expression was down-regulated, and ACSL4 expression in astrocytes and Schwann cells of DRGs was up-regulated in NP group ( P<0.05). Compared with NP group, MWT was significantly increased and TWL was prolonged at T 3-6, levels of iron ions, ROS and MDA in DGRs were decreased, activities of SOD and GSH-Px were increased, ACSL4 expression was down-regulated, GPX4 expression was up-regulated, and ACSL4 expression in astrocytes and Schwann cells of DRGs was down-regulated ( P<0.05), and no significant change was found in the parameters mentioned above in NP+ Veh group ( P>0.05). The results of electron microscopy showed that collapsed mitochondrial cristae and membrane rupture were found in astrocytes and Schwann cells of DRGs in NP group, and the number of collapsed mitochondrial cristae and membrane rupture was significantly decreased in NP + Lip group when compared with NP group. Conclusions:The ferroptosis in DRGs is involved in NP in rats.

Objective:The purpose of this study was to investigate the expression and role of N-myc downstream regulatory gene 2 (NDRG2) in radiation resistance of bladder cancer cells.Methods:T24 cells were cultured in vitro and irradiated with different doses of X-ray (0, 2, 4, 8, 10 and 20 Gy). The best dose of X-ray was selected for subsequent treatment. The radioresistant BCa cell line T24/R was established. The cytotoxicity of T24/R cells was detected by counting kit-8 (CCK-8) method. The proliferation and invasion ability of T24/R cells and T24 cells were detected by flow cytometry and transwell, respectively. Western blot was used to detect the expression of epithelial mesenchymal transition (EMT) related proteins. The survival rate of T24/R group (control group) and T24/R-NDRG 2 group was detected, and the migration ability of T24/R-NDRG 2 cells was detected after 2 Gy treatment. Results:The cell viability was inhibited significantly when the dose of X-ray was ≥2 Gy X-ray, so 2 Gy X-ray irradiation was chosen as the best condition for BCa cytotoxicity and T24/R radiation resistance cell line was successfully established; Apoptosis test showed that the number of S-phase cells was increased in T24/R group, and the proportion of S-phase cells in T24/R vs T24 was (26.49±4.5)% vs (14±2.6)% ( P<0.05); Transwell test showed that T24/R cells showed stronger migration ability than control group ( P<0.05), but there was no significant difference in EMT related protein expression between the two groups ( P>0.05). Overexpression of NDRG2 can significantly decreased the activity and migration ability of radiation-resistant T24/R cells ( P<0.05) when the radiation dose was gradually increasing in both groups. Conclusions:The radiation resistance of BCa cells is one of the causes of local tumor recurrence. Up-regulation of NDRG2 expression can inhibit the radiation resistance of T24 cells, so it can be used as a candidate for treatment of radiation-resistant BCa patients.

Objective:To investigate the effects of osimertinib on proliferation, migration and invasion of procollagen-lysine 2-oxoglutarate 5-dioxygenase 2 (PLOD2) overexpressing HCC827 cells and explore the potential mechanism of PLOD2 induced osimertinib resistance.Methods:We transfected HCC827 cells with LV-vector and LV-over/PLOD2. The expression of PLOD2 was detected by quantitative real time polymerase chain reaction (qRT-PCR) and western blotting. The effects of osimertinib on the proliferation of HCC827-vector and HCC827-PLOD2 cells were evaluated by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H tetrazolium bromide (MTT) assay. The effects of osimertinib on the migration and invasion of HCC827-vector and HCC827-PLOD2 cells were determined by Transwell assays. The expressions of E-cadherin and vimentin in cells were detected by immunofluorescence (IF). The expressions of epithelial-mesenchymal transition (EMT), FAK-PI3K/AKT and MAPK signal pathway related proteins were detected by western blotting.Results:The MTT assay showed that HCC827-PLOD2 cells were hyposensitive to osimertinib. The 50% inhibitory concentration (IC 50) and resistance index of osimertinib for HCC827-PLOD2 cells was over 1 000 nmol/L and over 100, respectively. The result of wound healing assay showed that the migration distance of HCC827-PLOD2 was about (2.13±0.21) fold changes as that of HCC827-vector cells. The result of Transwell assay showed that the numbers of HCC827-PLOD2 passing through the matrix membrane were (212.78±10.43), significantly higher than (101.32±12.52) of HCC827-vector cells ( P<0.01). The result of IF showed that compared with HCC827-vector cells, the expression of E-cadherin was down-regulated while vimentin was up-regulated in HCC827-PLOD2 cells. Osimertinb downregulated E-cadherin and upregulated vimentin expression in HCC827-vector cells but had limited effect in HCC827-PLOD2 cells. The result of western blotting showed that PLOD2 significantly increased vimentin expression level while decreased E-cadherin expression level. Osimertinib inhibited the expression of p-EGFR, but did not affect the expressions of PLOD2, p-FAK, p-AKT, p-ERK, vimentin and E-cadherin in HCC827-PLOD2 cells. Conclusion:PLOD2 confers resistance to osimertinib in HCC827 cells by regulating EMT, FAK-PI3K/AKT and MAPK signal pathways.

Objective:To investigate the effects of osimertinib on proliferation, migration and invasion of procollagen-lysine 2-oxoglutarate 5-dioxygenase 2 (PLOD2) overexpressing HCC827 cells and explore the potential mechanism of PLOD2 induced osimertinib resistance.Methods:We transfected HCC827 cells with LV-vector and LV-over/PLOD2. The expression of PLOD2 was detected by quantitative real time polymerase chain reaction (qRT-PCR) and western blotting. The effects of osimertinib on the proliferation of HCC827-vector and HCC827-PLOD2 cells were evaluated by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H tetrazolium bromide (MTT) assay. The effects of osimertinib on the migration and invasion of HCC827-vector and HCC827-PLOD2 cells were determined by Transwell assays. The expressions of E-cadherin and vimentin in cells were detected by immunofluorescence (IF). The expressions of epithelial-mesenchymal transition (EMT), FAK-PI3K/AKT and MAPK signal pathway related proteins were detected by western blotting.Results:The MTT assay showed that HCC827-PLOD2 cells were hyposensitive to osimertinib. The 50% inhibitory concentration (IC 50) and resistance index of osimertinib for HCC827-PLOD2 cells was over 1 000 nmol/L and over 100, respectively. The result of wound healing assay showed that the migration distance of HCC827-PLOD2 was about (2.13±0.21) fold changes as that of HCC827-vector cells. The result of Transwell assay showed that the numbers of HCC827-PLOD2 passing through the matrix membrane were (212.78±10.43), significantly higher than (101.32±12.52) of HCC827-vector cells ( P<0.01). The result of IF showed that compared with HCC827-vector cells, the expression of E-cadherin was down-regulated while vimentin was up-regulated in HCC827-PLOD2 cells. Osimertinb downregulated E-cadherin and upregulated vimentin expression in HCC827-vector cells but had limited effect in HCC827-PLOD2 cells. The result of western blotting showed that PLOD2 significantly increased vimentin expression level while decreased E-cadherin expression level. Osimertinib inhibited the expression of p-EGFR, but did not affect the expressions of PLOD2, p-FAK, p-AKT, p-ERK, vimentin and E-cadherin in HCC827-PLOD2 cells. Conclusion:PLOD2 confers resistance to osimertinib in HCC827 cells by regulating EMT, FAK-PI3K/AKT and MAPK signal pathways.

Objective: To investigate the impact of n-3 polyunsaturated fatty acid (n-3 PUFA) on function and expression of store-operated calcium channels (SOCC) in coronary artery smooth muscle cells (SMC) derived from diabetic rat. Methods: A total of 180 healthy male Sprague-Dawley (SD) rats were randomly divided into normal group (N, n=45), placebo-treated diabetic group (D, n=45), lose dose n-3 PUFA treated diabetic group (DL, n=45) and high dose n-3 PUFAs treated diabetic group (DH, n=45). Streptozotocin-induced diabetic rat animal model was established by two consecutive intraperitoneal injections. After modeling, rats in group DL and DH were treated with 10 mg·kg^-1·d^-1 and 50 mg·kg^-1·d^-1 n-3 PUFAs respectively per gavage for eight weeks. After eight weeks, rat coronary artery SMC was isolated by enzyme digestion. Changes of cytosolic calcium concentration in coronary artery SMC were examined by calcium fluorescence imaging technique, coronary artery tension was detected by myograph system, and protein expressions of SOCC on coronary artery SMC were measured by Western blot. Results: SOCC induced ΔF340/F380 of group N, D, DL and DH were 0.425±0.023, 0.838±0.037, 0.342±0.052 and 0.364±0.045 respectively, which was significantly lower in group N, DL, DH than in group D (P<0.05). SOCC induced changes of tensions were 0.94±0.09, 1.95±0.18, 1.35±0.24 and 1.01±0.18 in the group N, D, DL and DH, respectively, which was significantly lower in group N and DH than in group D (P<0.05). Protein expressions of STIM1, Orai1 and TRPC1 were significantly higher in diabetic rat coronary SMC than in group N (P<0.05). STIM1 protein expressions were significantly lower in group DL and DH than in group D, and Orai1 and TRPC1 protein expressions were similar among group. Conclusions Coronary artery tension, cytosolic calcium concentration and protein expressions of SOCC are higher in diabetic rat coronary artery SMC when compared with normal rats. n-3 PUFA intervention could downregulate the protein expression of SOCC, reduce cytosolic calcium concentration and coronary artery tension, and is protective to the diabetic injury in coronary artery.

Objective To investigate the impact of n?3 polyunsaturated fatty acid (n?3 PUFA) on function and expression of store?operated calcium channels (SOCC) in coronary artery smooth muscle cells (SMC) derived from diabetic rat. Methods A total of 180 healthy male Sprague?Dawley (SD) rats were randomly divided into normal group (N, n=45), placebo?treated diabetic group (D, n=45), lose dose n?3 PUFA treated diabetic group (DL, n=45) and high dose n?3 PUFAs treated diabetic group (DH, n=45). Streptozotocin?induced diabetic rat animal model was established by two consecutive intraperitoneal injections. After modeling, rats in group DL and DH were treated with 10 mg·kg-1·d-1 and 50 mg·kg-1·d-1 n?3 PUFAs respectively per gavage for eight weeks. After eight weeks, rat coronary artery SMC was isolated by enzyme digestion. Changes of cytosolic calcium concentration in coronary artery SMC were examined by calcium fluorescence imaging technique, coronary artery tension was detected by myograph system, and protein expressions of SOCC on coronary artery SMC were measured by Western blot. Results SOCC induced ΔF340/F380 of group N, D, DL and DH were 0.425±0.023, 0.838±0.037, 0.342±0.052 and 0.364± 0.045 respectively, which was significantly lower in group N, DL, DH than in group D (P<0.05). SOCC induced changes of tensions were 0.94±0.09, 1.95±0.18, 1.35±0.24 and 1.01±0.18 in the group N, D, DL and DH, respectively, which was significantly lower in group N and DH than in group D (P<0.05). Protein expressions of STIM1, Orai1 and TRPC1 were significantly higher in diabetic rat coronary SMC than in group N (P<0.05). STIM1 protein expressions were significantly lower in group DL and DH than in group D, and Orai1 and TRPC1 protein expressions were similar among group. Conclusions Coronary artery tension, cytosolic calcium concentration and protein expressions of SOCC are higher in diabetic rat coronary artery SMC when compared with normal rats. n?3 PUFA intervention could downregulate the protein expression of SOCC, reduce cytosolic calcium concentration and coronary artery tension, and is protective to the diabetic injury in coronary artery.

OBJECTIVE:To establish a method for the simultaneous determination of gallic acid,rhein and emodin in Com-pound xiaozhi suppository. METHODS:HPLC was performed on the column of Kromasil C18 with mobile phase of 0.5% phosphor-ic acid-Acetonitrile solution(gradient elution)at a flow rate of 1.0 ml/min,the detection wavelength was 273 nm for gallic acid and 254 nm for rhein and emodin,the column temperature was 28 ℃,and the injection volume was 20 μl. RESULTS:The linear range was 0.296 4-14.82 μg/ml for gallic acid(r=0.999 6),0.215 0-10.75 μg/ml for rhein(r=0.999 9)and 0.307 2-15.36 μg/ml for emo-din(r=0.999 9);RSDs of precision,stability and reroducibility tests were lower than 3.0%;recoveries were 95.1%-97.2%(RSD=0.64%,n=6),95.4%-97.2%(RSD=0.42%,n=6) and 96.5%-99.4%(RSD=1.10%,n=6),respectively. CONCLU-SIONS:The method is simple and accurate,and can be used for the contents determination of gallic acid,rhein and emodin in Compound xiaozhi suppository.