OBJECTIVES: To evaluate the protective effect of Schistosoma japonicum cystatin (rSj-Cystatin) in a mouse mode of "two-hit" sepsis. METHODS: Sixty male C57BL/6 mice randomized equally into sham-operated group, protein group, "two-hit" modeling group, and protein intervention group. In the former two groups, the mice received an intraperitoneal injection of 100 μL PBS followed by exposure of the cecum and then by intraperitoneal injection of 100 μL PBS or 25 μg rSj-Cystatin 30 min later; In the latter two groups, 100 μL PBS containing LPS (5 mg/kg) was injected intraperitoneally 24 h before cecal ligation and puncture (CLP), and 100 μL PBS or 25 μg rSj-Cystatin were injected 30 min after CLP. At 12 h after rSj-Cystatin treatment, 6 mice from each group were sacrificed for detection of TNF-α, IL-6, IL-10, TGF-β, iNOS and Arg-1 in the serum, spleen, liver, lung and kidney tissues using ELISA, for examinations of liver, lung and kidney pathologies with HE staining, and for analysis of CD3⁺CD4⁺CD25⁺Foxp3⁺ T cell percentage in the spleen using flow cytometry. The remaining mice were observed for general condition and 72-h survival. RESULTS: The 72-h survival rates in the 4 groups were 100%, 100%, 0% and 20%, respectively, showing significant differences between the latter two groups. The mouse models of "two-hit" sepsis exhibited obvious tissue pathologies and significant elevations of TNF-α and IL-6 in both the serum and tissue homogenate, which were significantly ameliorated by rSj-Cystatin treatment. Treatment with rSj-Cystatin also increased IL-10 and TGF-β levels and spleen CD3⁺CD4⁺CD25⁺Foxp3⁺ T cell percentage. The septic mouse models also showed increased iNOS levels in all the detected tissues and a decreased Arg-1 level in the kidney, and these changes were obviously improved by rSj-Cystatin treatment. CONCLUSIONS rSj-Cystatin has a protective effect against "two-hit" sepsis in mice by regulating the inflammatory microenvironment.
Animals ; Mice ; Sepsis/drug therapy* ; Male ; Schistosoma japonicum/chemistry* ; Mice, Inbred C57BL ; Cystatins/therapeutic use* ; Interleukin-10/metabolism* ; Interleukin-6/blood* ; Tumor Necrosis Factor-alpha/blood* ; Disease Models, Animal ; Transforming Growth Factor beta/metabolism*

OBJECTIVE To investigate the effect and mechanism of miR-152-3p on the resistance to paclitaxel(PTX)of PTX-resistant ovarian cancer cells(A2780T cells).METHODS ① Ovarian cancer parent cells(A2780 cells)and A2780T cells were treated with PTX(1.875,3.75,7.5,17 and 23 μmol·L-1)for 48 h.Cell viability was evaluated by MTT assay,and the 50%inhibitory concentration(IC50)and drug resistance index of A2780T cells were calculated.Western blotting was used to detect the expres-sions of resistance protein P-glycoprotein(P-gp),multidrug resistance related protein 1(MRP1)and adenosine triphosphate binding transporter G superfamily member 2(ABCG2).② Real-time fluorescent quantita-tive PCR(RT-qPCR)was used to detect the expressions of miR-152-3p in A2780 and A2780T cells.The lipid-mediated transient transfection technique was employed to transfect the miR-152-3p inhibitor to reduce miR-152-3p expression in A2780T cells(miR-152-3p inhibitor group),while the negative control(miR-152-3p NC)group was established.RT-qPCR was used to detect transfection efficiency,and the MTT method,scratch experiment,and flow cytometry were used to investigate the effects of the trans-fecting miR-152-3p inhibitor on survival,migration and apoptosis of A2780T cells.Western blotting was used to detect the protein expressions of Bax and Bcl-2 in A2780T cells.③ Bioinformatics analysis of databases including miRDB,Targetscan,miRWalk,and Starbase predicted the target genes of miR-152-3p that were verified by Western blotting to detect the protein expression of PTEN in A2780T cells of the miR-152-3p inhibitor and miR-152-3p NC groups,and RT-qPCR to detect the PTEN mRNA expression in A2780 and A2780T cells.Then,the lipid-mediated transient transfection technique was used to transfect PTEN siRNA to silence PTEN expression in A2780T cells(PTEN siRNA group).The siRNA negative control(siRNA NC)group was established.RT-qPCR was used to detect transfection efficiency,the MTT method was employed to measure the survival rate and IC50 value,and Western blotting was used to assess the protein expressions of P-gp,MRP1,and ABCG2 in A2780T cells after silencing PTEN expression.RESULTS ①After treatment with PTX,the cell survival rates were decreased in A2780 and A2780T cells(P<0.05),and the resistance index of A2780T cells was 2.8.Compared with A2780 cells,the protein expressions of P-gp and MRP1 and ABCG2 were highly expressed in A2780T cells(P<0.05,P<0.01).② RT-qPCR showed that the expression of miR-152-3p in A2780T cells was higher than that of A2780 cells(P<0.01).Compared with the miR-152-3p NC group,A2780T cell viability(P<0.05,P<0.01)and cell migration capability(P<0.05)were significantly inhibited,while the apoptosis rate increased(P<0.01)in miR-152-3p inhibitor group.Moreover,the protein expression of Bax was increased(P<0.01),but Bcl-2 decreased(P<0.05).③ Bioinformatics analysis suggested that PTEN was a target gene of the miR-152-3p,and the verified results showed that the PTEN protein expression in A2780T cells of the miR-152-3p inhibitor group was lower than that of the miR-152-3p NC group(P<0.05),and PTEN mRNA expression in A2780T cells was higher than that in A2780 cells(P<0.01).After silencing the expression of PTEN in A2780T cells,the cell viability was significantly reduced(P<0.05,P<0.01),while the IC50 value was reduced(P<0.01)compared with the siRNA NC group.In addition,the protein expressions of P-gp,MRP1 and ABCG2 were decreased(P<0.05,P<0.01).CONCLUSION miR-152-3p is highly expressed in A2780T cells,and down-regulation of its expression may inhibit proliferation and migration,prompt apoptosis and reduce the resistance to PTX of A2780T cells,which is made possible by inhibiting expression of its target gene PTEN.

{L-End}Objective To analyze the characteristics of hearing loss and the influencing factors of high-frequency hearing loss (HFHL) among noise-exposed workers in an urban rail transit enterprise over five consecutive years. {L-End}Methods A total of 1 268 noise-exposed workers, who exposed to the average noise intensity of <85.0 dB(A), in an urban rail transit enterprise was selected as the research subjects using a judgment sampling method. The pure-tone audiometry results from 2019 to 2023 were collected to analyze the result of hearing loss. The influencing factors of HFHL (average hearing threshold ≥40.0 dB at high frequencies in both ears) were analyzed using the generalized estimating equations (GEE). {L-End}Results The detection rates of threshold elevations at frequencies of 0.5-6.0 kHz increased with increasing frequency from 2019 to 2023 (all P<0.01), with the highest detection rate at 6.0 kHz. The detection rate of speech frequency hearing loss (hearing threshold weighted value≥26.0 dB in the better ear) was 0.1%, 0.0%, 0.4%, 0.2%, and 0.2%, respectively. The detection rate of HFHL from 2019 to 2023 was 2.4%, 2.8%, 2.8%, 2.1%, and 2.8%, respectively. The GEE analysis results showed that the risk of HFHL of the workers in 2022 and 2023 was lower than that in 2019 (all P<0.01), with the odds ratios and 95% confidence intervals ［OR (95%CI)］ of 0.57 (0.41-0.81) and 0.65 (0.48-0.87), respectively. The risk of HFHL was higher among vehicle maintenance worker than train drivers (P<0.05), with OR (95%CI) of 2.37 (1.18-4.77). The risk of HFHL increased with age and length of service among the workers (all P<0.05), with the OR (95%CI) of 2.05 (1.22-3.46) and 1.69 (1.12-2.54), respectively. No interaction was found between type of job and age, type of job and length of service, or age and length of service in the risk of HFHL among the research subjects(all P<0.05). {L-End}Conclusion Noise exposure below the national occupational exposure limits can lead to hearing loss in noise-exposed workers of urban rail transit enterprises, possibly affecting the hearing threshold at 6.0 kHz first. The influencing factors for HFHL in workers of rail transit are age, length of service, and type of job. There is a dose-effect relationship with age and length of service.

Objective:To compare the clinical curative effect of roxadustat and recombinant human erythropoietin (rhEPO) on non-dialysis stage 3 to 5 chronic kidney disease (CKD) combined with renal anemia (RA).Methods:A total of 108 patients with non-dialysis stage 3 to 5 CKD and RA admitted to Jiangbei Branch of Zhongda Hospital Affiliated to Southeast University were prospectively enrolled between January 2020 and October 2022. According to random number table method, they were divided into group A and group B, 54 cases in each group. The group A was treated with roxadustat and polysaccharide-iron complex (PIC) for 3 months, while group B was treated with rhEPO and PIC for 3 months. The clinical curative effect, levels of hemoglobin (Hb), red blood cell count (RBC), hematocrit (Hct), serum ferritin (SF), transferrin saturation (TSAT), transferrin (TRF), blood urea nitrogen (BUN), serum creatinine (Scr) and β2-microglobulin (β2-MG) before and after treatment, and the incidence of adverse reactions during treatment in the two groups were compared.Results:The total response rate of treatment in group A was significantly higher than that in group B: 87.04% (47/54) vs. 70.37% (38/54), and the difference was statistically significant ( P<0.05). After treatment, levels of RBC, Hb and Hct in group A were significantly higher than those in group B: (3.47 ± 0.59) × 10 12/L vs. (2.60 ± 0.51) × 10 12/L, (110.45 ± 12.97) g/L vs. (93.64 ± 10.58) g/L, 0.358 ± 0.054 vs. 0.303 ± 0.043, and the difference was statistically significant ( P<0.05). The levels of TSAT, SF and TRF in group A were significantly higher than those in group B: (35.17 ± 3.65)% vs. (29.82 ± 3.10)%, (286.74 ± 17.23) μg/L vs. (243.16 ± 15.49) μg/L, (2.76 ± 0.45) g/L vs. (2.40 ± 0.32) g/L, and the difference was statistically significant ( P<0.05). The levels of BUN, Scr and β2-MG in group A were significantly lower than those in group B: (3.98 ± 0.41) mmol/L vs. (4.36 ± 0.54) mmol/L, (62.57 ± 7.89) μmol/L vs. (80.34 ± 9.65) μmol/L, (1.50 ± 0.42) μg/L vs. (1.99 ± 0.58) μg/L, and the difference was statistically significant ( P<0.05). During treatment, incidence of adverse reactions in group A was significantly lower than that in group B: 11.11% (6/54) vs. 25.93% (14/54), and the difference was statistically significant ( P<0.05). Conclusions:Compared with rhEPO, roxadustat has better curative effect, which can effectively relieve anemia and improve iron metabolism in patients with non-dialysis stage 3 to 5 CKD and RA, with good safety.

Objective:To investigate the value of plasma Epstein-Barr virus (EBV) DNA detection in the screening of nasopharyngeal carcinoma (NPC) and its clinical application in non-high-risk areas.Methods:Plasma EBV DNA results in 1 153 newly diagnosed nasopharyngeal carcinoma patients who were treated in Sichuan Cancer Hospital from 2015 to 2020 and 244 healthy control cases with matched sex and age were retrospectively analyzed. EBV DNA were detected by quantitative real-time PCR. Positive rate of EBV DNA was determined by the cutoff value of 400 (≥400 copies/ml as positive) and optimization threshold method (presence of S amplification curve as positive). Further analyses were conducted to compare EBV DNA load in different clinical stage, TNM stage and regions distribution characteristics. Receiver operating characteristic (ROC) curve analysis was performed to evaluate the diagnostic value of the cutoff value of 400 and optimization threshold method for NPC.Results:Compared with healthy controls, EBV DNA increased significantly in newly diagnosed NPC patients ( P<0.001). Both evaluation methods revealed that the EBV DNA positive percentage increased with TNM and clinical stage ( P<0.001). With 400 copies/ml as cutoff value, the diagnostic sensitivity and specificity were 40.85% and 100%, respectively. The area under the curve was 0.704 (95% CI 0.676-0.733, P<0.001). Evaluated by the optimization threshold method, the sensitivity and specificity could improve to 82.0% and 99.2%, respectively, and the area under the curve reached 0.910 (95% CI 0.894-0.924, P<0.001). Conclusions:In the low prevalence area of nasopharyngeal carcinoma, the sensitivity for diagnosis of nasopharyngeal carcinoma is only 40.9% by the 400 copies/ml cutoff value method. The optimization threshold method is a better choice to improve the diagnostic sensitivity without lowering the diagnostic specificity.

Objective:To analyze the clinical data of children with active lupus nephritis(LN) with poor first-line treatment and further treatment with belimumab, and explore the efficacy and safety of belimumab in the treatment of children with LN, so as to provide experience and guidance for clinical treatment of children with LN.Methods:From August 2020 to September 2021, 12 children with LN whose systemic lupus erythematosus disease activity index(SLEDAI)score was ≥8 and with poor first-line treatment were collected, and their clinical manifestations, treatment process, SLEDAI score, complement C3, complement C4, anti-dsDNA antibody titer, and proteinuria relief were analyzed retrospectively.Results:Before treatment with belimumab, the SLEDAI score was 8 in 3 cases, 10 in 5 cases, 12 in 2 cases and 16 in 2 cases.Theurine protein was positive in 6 cases.The anti-dsDNA antibody titer was higher than normal value in 8 cases.The complement C3 decreased in 8 cases and the complement C4 decreased in 6 cases.The SLEDAI scores and the anti-dsDNA antibody of 12 children and 24-hour urine protein quantification of 6 children with positive urine protein began to decrease within 4 weeks after treatment with belimumab.Anti-dsDNA antibody decreased to normal in 12th week and 24 h urine protein decreased to normal in 16th week.The levels of complement C3 and C4 began to rise within 4 weeks, complement C3 returned to normal within 24 weeks, and complement C4 returned to normal within 28 weeks.Conclusion:For LN children with poor response to first-line therapy or persistent disease activity, the addition of belimumab resulted in increased complement, decreased disease activity index and anti-dsDNA antibody titer, and effective relief of proteinuria.The application of belimumab has a certain effect on active LN children with poor response to first-line therapy, which is worthy of clinical promotion.

Objective:To preliminarily investigate the efficacy and safety of bevacizumab plus neoadjuvant chemotherapy combined with concurrent chemoradiotherapy in the treatment of locally advanced giant cervical cancer (LACC).Methods:Twenty-four patients with LACC who were treated with bevacizumab combined with neoadjuvant chemotherapy were assigned into the study group, and 30 patients receiving neoadjuvant chemotherapy in phase Ⅱ clinical trial (ChiCTR-TRC-11001832) were allocated in the control group. The survival rate was calculated by the Kaplan- Meier method, and the significance of differences between the variables was determined by the log-rank test. Results:The tumor volumes were (1.64±23.15) cm 3 and (12.83±15.08)( P=0.037), and the complete remission (CR) rates were calculated as 45.8% and 13.3%( P=0.004) in the study and control groups after neoadjuvant chemotherapy. The tumor volumes were (0±1.5) cm 3 and (1.00±10.63) cm 3( P=0.022) and the CR rates were 70% and 50%( P=0.009) in the study and control groups before afterloading treatment. The median follow-up was 24.6(9.3-101.7) months. The 1-and 2-year overall survival rates were 96%, 96% and 90%, 71%( P=0.110), the recurrence-free survival rates was 96%, 96% and 97%, 89%( P=0.512), and the distant metastasis-free survival rates were 96%, 88% and 83%, 80%( P=0.297) in the study and control groups, respectively. Adverse reactions were acceptable in both groups. Conclusion:Bevacizumab combined with neoadjuvant chemotherapy can significantly reduce the tumor volume, improve the tumor CR rate and yield tolerable adverse reactions.

Objective:To observe the effect of ankle stretching on ankle biomechanics, balance, walking ability and ability in the activities of daily living among stroke survivors.Methods:Eighteen hemiplegic stroke survivors were randomly divided into an experimental group ( n=9) and a control group ( n=9). In addition to routine medication and rehabilitation training, the experimental group received 20 minutes of ankle joint stretching daily while the control group underwent an additional twenty minutes of routine rehabilitation training. Before and after the treatment, both groups′ ankle joint stiffness (K), muscle strength, active range of motion (AROM) and passive range of motion (PROM) were evaluated. They were also assessed using the modified Ashworth scale (MAS), the Fugl-Meyer lower extremity assessment (FMA-LE), the Berg balance scale (BBS), the 6-minute walking test (6MWT) and the modified Barthel Index (MBI). Results:After two weeks of treatment significant improvement was observed in the AROM and muscle strength of both groups in dorsiflexion and plantarflexion. The average BBS and FMA-LE scores of both groups had also improved significantly. Significant improvement in the average PROM of plantarflexion and the K of dorsiflexion, as well as in average MBI score was observed only in the treatment group. After two weeks the treatment group′s average muscle strength in plantarflexion and dorsiflexion was significantly better than the control group′s.Conclusions:Stretching can reduce ankle stiffness, improve the range of motion, muscle strength, and ability of in the activities of daily living after a stroke.

Chondroitin sulfate (CS) is a sulfurated glycosaminoglycan, a major component of the extracellular matrix, widely distributed in skin, cartilage and vascular tissue. CS plays an important role in the physiological state regulation of articular cartilage, which affects tensile strength and elasticity of tissues by influencing aggrecan. Previous studies have shown that CS sulfate modification may be related to the growth and development disorders of cartilage tissue and the occurrence of osteoarticular diseases. At the same time, CS is also a common joint supplement, often used in the treatment of osteoarthritis and Kashin-Beck disease. In this paper, the research progress of CS sulfate modification characteristics in Kashin-Beck disease and osteoarthritis and the application of the preparation in the treatment of Kashin-Beck disease and osteoarthritis are reviewed, aiming to provide help for the investigation of the etiology of Kashin-Beck disease and the treatment of osteoarthritis and Kashin-Beck disease.

Objective:To analyze the effect of systematic exercise with a self-designed foot-ankle exerciser on postoperative ankle function in patients with tibial pilon fracture.Methods:A retrospective study was conducted of the 84 patients with tibial pilon fracture who had been treated by open reduction and internal fixation at Nanhua Hospital Affiliated to Nanhua University from May 2018 to January 2020. According to the difference in rehabilitation, they were divided into a control group ( n=42) which were subjected to staged functional rehabilitation after operation and an observation group ( n=42) which was subjected to systematic exercise with a self-designed foot-ankle exerciser. In the control group there was 26 males and 16 females with an age of (36.6±4.8) years (from 20 to 55 years); in the observational group there were 27 males and 15 females with an age of (36.4±4.5) years (from 21 to 57 years). The 2 groups were compared in terms of postoperative recovery, ankle functional recovery, postoperative quality of life, and rate of complications. Results:The 2 groups were comparable due to the insignificant differences in their preoperative general data ( P>0.05). The time for first ambulation [(3.7±0.3) weeks], time for fracture union [(9.1±0.8) weeks], and time for full-weight bearing [(6.5±0.9) weeks] in the observational group were all significantly shorter than those in the control group [(4.1±0.5) weeks, (10.9±1.2) weeks and (7.2±1.1) weeks] ( P<0.05). The good to excellent rate of ankle functional recovery was 92.9% (39/42) for the observational group, significantly higher than that for the control group (73.8%, 31/42) ( P<0.05). The scores for postoperative quality of life in the observational group were significantly higher than those in the control group in dimensions of material, psychology, society and body ( P<0.05). The rate of total postoperative complications in the observational group was 2.4% (1/42), significantly lower than that in the control group (10.9%, 8/42) ( P<0.05). Conclusions:For patients with tibial pilon fracture undergoing the same surgical procedures, systematic exercise with our self-designed foot-ankle exerciser can promote functional recovery of the ankle joint, improve postoperative quality of life, and reduce incidence of postoperative complications.