Mechanism of large-conductance calcium-activated potassium channel involved in inflammatory response in sepsis / 中华危重病急救医学

OBJECTIVE@#To explore the mechanisms of large-conductance calcium-activated potassium channel (BKCa) involved in inflammatory response in sepsis.@*METHODS@#The serum levels of BKCa were measured by enzyme-linked immunosorbent assay (ELISA) in patients with sepsis (28 cases), patients with common infection (25 cases) and healthy people (25 cases). The relationship between levels of BKCa and acute physiology and chronic health evaluation II (APACHE II) were analyzed. Cultured RAW 264.7 cells were stimulated by lipopolysaccharide (LPS). In some experiments, a cell model of sepsis was constructed using Nigericin as the second stimulus signal. The mRNA and protein expressions of BKCa in RAW 264.7 cells stimulated with LPS (0, 50, 100, 1 000 μg/L) were measured by real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) and Western blotting. RAW 264.7 cells were transfected with small interfering RNA of BKCa (siRNA-BKCa), and the levels of caspase-1 precursor (pro-caspase-1), interleukin-1β precursor (pro-IL-1β) in cell, and the levels of caspase-1 p20, IL-1β p17 of cell culture medium, and NOD-like receptor protein 3 (NLRP3), nuclear factor-κB (NF-κB) were measured by Western blotting. The apoptosis were detected by staining with propidium iodide (PI), the release rate of lactate dehydrogenase (LDH) were measured, and the expression of apoptotic protein Gasdermin D (GSDMD) was measured by Western blotting to evaluate the effect of silencing BKCa on cell pyrosis.@*RESULTS@#The level of serum BKCa in patients with sepsis was significantly higher than that in patients with common infection and health peoples (ng/L: 165.2±25.9 vs. 102.5±25.9, 98.8±20.0, both P < 0.05). In addition, the level of serum BKCa in patients with sepsis was significantly positively correlated with APACHE II score (r = 0.453, P = 0.013). LPS could construct a sepsis cell model by which LPS could promote BKCa expression in mRNA and protein with a concentration-dependent manner. The mRNA and protein expressions of BKCa in the cells stimulated by 1 000 μg/L LPS were significantly higher than that in the blank group (0 μg/L) [BKCa mRNA (2-ΔΔCt): 3.00±0.36 vs. 1.00±0.16, BKCa/β-actin: 1.30±0.16 vs. 0.37±0.09, both P < 0.05]. Compared with the control group, the ratios of caspase-1 p20/pro-caspase-1 and IL-1β p17/pro-IL-1β in the model group were significantly increased (caspase-1 p20/pro-caspase-1: 0.83±0.12 vs. 0.27±0.05, IL-1β p17/pro-IL-1β: 0.77±0.12 vs. 0.23±0.12, both P < 0.05), however, transfection of siRNA-BKCa induced the decrease both of them (caspase-1 p20/pro-capase-1: 0.23±0.12 vs. 0.83±0.12, IL-1β p17/pro-IL-1β: 0.13±0.05 vs. 0.77±0.12, both P < 0.05). Compared with the control group, the number of apoptotic cells, LDH release rate and GSDMD expression in the model group were significantly increased [LDH release rate: (30.60±8.40)% vs. (15.20±7.10)%, GSDMD-N/GSDMD-FL: 2.10±0.16 vs. 1.00±0.16, both P < 0.05], however, transfection of siRNA-BKCa induced the decrease both of them [LDH release rate: (15.60±7.30)% vs. (30.60±8.40)%, GSDMD-N/GSDMD-FL: 1.13±0.17 vs. 2.10±0.16, both P < 0.05]. The mRNA and protein expressions of NLRP3 in sepsis cells were significantly higher than those in the control group [NLRP3 mRNA (2-ΔΔCt): 2.06±0.17 vs. 1.00±0.24, NLRP3/GAPDH: 0.46±0.05 vs. 0.15±0.04, both P < 0.05]. However, the expression of NLRP3 after siRNA-BKCa transfection was significantly lower than that in model group [NLRP3 mRNA (2-ΔΔCt): 1.57±0.09 vs. 2.06±0.17, NLRP3/GAPDH: 0.19±0.02 vs. 0.46±0.05, both P < 0.05]. Compared with the control group, the NF-κB p65 nuclear transfer of sepsis cell were significantly increased (NF-κB p65/Histone: 0.73±0.12 vs. 0.23±0.09, P < 0.05). However, the NF-κB p65 expression in the nucleus were decreased after siRNA-BKCa transfection (NF-κB p65/Histone: 0.20±0.03 vs. 0.73±0.12, P < 0.05).@*CONCLUSIONS@#BKCa is involved in the pathogenesis of sepsis, and its possible mechanism is to activate NF-κB/NLRP3/caspase-1 signaling pathway to induce inflammatory factor production and cell death.

Effects of apatinib on quality of life and immune function in older adult patients with advanced non-small cell lung cancer / 中国基层医药

Objective:To analyze the effects of apatinib on quality of life and immune function in older adult patients with advanced non-small cell lung cancer.Methods:A total of 187 older adult patients with advanced non-small cell lung cancer admitted to Taizhou Cancer Hospital from January 2017 to January 2021 were included in this study. They were divided into the control group ( n = 93) and the observation group ( n = 94). The control group was treated with carboplatin combined with pemetrexed and the observation group was treated with apatinib based on carboplatin and pemetrexed. Sign and symptoms remission was compared between the observation and control groups. The levels of tumor markers, immune function, and quality of life score were compared between the two groups before and after treatment. Results:Total remission rate in the observation group was significantly higher than that in the control group (88.30% vs. 69.89%, χ2 = 9.59, P < 0.05). After treatment, carbohydrate antigen 125, carbohydrate antigen 50, and carcinoembryonic antigen in the observation group were (16.25 ± 5.47) μg/L, (15.23 ± 3.27) μg/L and (5.91 ± 2.66) mg/L, respectively, which were significantly lower than (21.49 ± 6.61) μg/L, (19.11 ± 3.48) μg/L and (10.14 ± 2.73) mg/L in the control group ( t = 5.91, 7.86, 10.73, all P < 0.05). The percentage of CD3 + and CD4 + cells, and the ratio of CD4 +/CD8 + cells in the observation group were (69.34 ± 8.85)%, (38.15 ± 6.52)%, (1.40 ± 0.33), respectively, which were significantly higher than (64.51 ± 8.74)%, (33.55 ± 6.33)%, (1.23 ± 0.25) in the control group ( t = -3.75, -5.36, -3.97, all P < 0.05). Quality of life score was increased in each group ( P < 0.001). The amplitude of increase in quality of life score was greater in the observation group compared with the control group ( P < 0.001). Conclusion:Apatinib can effectively reduce the level of tumor markers and improve immune function in older adult patients with advanced non-small cell lung cancer and improve quality of life.

Preliminary Application of Lauromacrogol in the Treatment of Varicose Veins of Lower Limb / 现代生物医学进展

Objective:To investigate the application value of lauromacrogol in the treatment of varicose veins of lower limb.Methods:The medical records of 75 patients (75 affected limbs) with varicosis of great saphenous vein of lower limbs confirmed between September 2013 and September 2015 were analyzed retrospectively.According to the treatment regimen,the patients were divided into two groups.Among them,33 cases (33 limbs) treated by ultrasound guided injection of foam sclerosing agent lauromacrogol were included in the lauromacrogol group while 42 cases (42 limbs) treated by high ligation of great saphenous vein combined with stripping were included in the operation group.The operation time,intraoperative blood loss,off-bed time,treatment costs and incidence of postoperative complications were recorded.Patients were followed up for 6 to 12 months after operation,and the recurrence rate was recorded.Results:The operation time,off-bed and length of hospital stay of the lauromacrogol group were shorter than those of the operation group;the intraoperative blood loss and hospitalization expenses were less than those of the operation group (P＜0.05);The incidence rates of subcutaneous hematoma and skin numbness in the lauromacrogol group were significantly lower than those in the operation group (P＜0.05);The 12-month recurrence rate in the lauromacrogol group (12.12％) was lower than that in the operation group (33.33％)(P＜0.05).Conclusion:Injection of foam sclerosing agent lauromacrogol is an alternative minimally invasive therapy for treating varicosis of great saphenous vein of lower limbs.

Establishment of a diet-induced obesity model in zebrafish larvae / 南方医科大学学报

<p><b>OBJECTIVE</b>To establish a diet-induced obesity model in zebrafish larvae.</p><p><b>METHODS</b>At 7 days post-fertilization (dpf), 200 zebrafish larvae with normal development were randomly allocated to two groups with the feeding quantity of 30 mg per day (normal feeding group) or 180 mg per day (overfed group) for 20 days. The weight, length, BMI, triglyceride (TG) and total cholesterol (TCH) of each group were measured. Whole-mount Oil Red O staining, frozen Oil Red O staining and hematoxylin-eosin (HE) staining were used to estimate the rate of hepatic steatosis and liver histology of the zebrafish. The dynamic change of hepatic lipid droplets and distribution of adipose tissue were observed with Nile Red staining in overfed zebrafish in vivo.</p><p><b>RESULTS</b>The weight, length, BMI and TG of overfed zebrafish were significantly increased compared with those in normal feeding group. Whole-mount Oil Red O staining showed that the percent of hepatic steatosis in overfed group (89.4%) was markedly higher than that in normal feeding group (20.7%). Macrovesicular steatosis was observed in the liver of the overfed larvae. Nile Red staining visualized hepatic lipid droplets and the distribution of larval adipose tissue, which increased with feeding time in the overfed zebrafish. Starving larvae showed depletion of fat and hepatic lipid, and adipose tissue was induced after refeeding.</p><p><b>CONCLUSIONS</b>We successfully established an diet-induced obesity model in zebrafish larva, in which Nile Red staining allows in vivo observation of the adipocytes and hepatic lipid droplets.</p>

The short-long term therapeutic effects of endovascular therapy for in-stent restenosis of superficial femoral artery / 中华普通外科杂志

Objective To evaluate therapeutic effects of endovascular therapy for in-stent restenosis of superficial femoral artery,and the risk factors of restenosis of re-intervention therapy.Methods We retrospectively summarized the clinical data of 35 patients(38limbs)who suffered from in-stent restenosis of superficial femoral artery(SFA)stent from 2010 to 2012.Results 35 patients (38 limbs)were enrolled in this study,there were 24 males and 11 females.Mean age was (68 ±8)years.The success rate of endovascular therapy was 89.5％ (34/38),including in-stent angioplasty in 14 limbs,in-stent and beyond-stent angioplasty in 5 limbs,in-stent angioplasty and beyond-stent re-stenting in 15 cases.The primary patency and sencondary patency rate for 6,12 and 24 months was 65.2％,46.5％,46.5％,and 87.5％,80.2％,55.8％ respectively.The independent risk factors for re-obliteration were age ＞ 70 years (P ＜ 0.05) and diabetes mellitus (P ＜ 0.05).Conclusions Endovascular therapy is effective and safe in treatment in-stent restenosis of superficial femoral artery.The long-term in-stent patency is not satisfactory,with age ＞ 70 years and diabetes mellitus as independent risk factors.