Role of pyroptosis in bilirubin-induced microglial injury / 中国当代儿科杂志

OBJECTIVE@#To study whether pyroptosis is involved in the bilirubin-induced injury of primary cultured rat cortical microglial cells.@*METHODS@#Primary cultured rat cortical microglial cells were randomly administered with 30 μmol/L bilirubin (bilirubin group), 30 μmol/L bilirubin following 30 μmol/L VX-765 pretreatment (VX-765+bilirubin group), or an equal volume of dimethyl sulfoxide (control group). Modified MTT assay was used to measure the viability of microglial cells. Western blot was used to measure the expression of the pyroptosis-related proteins Caspase-1 and gasdermin D (GSDMD). Lactate dehydrogenase (LDH)-release assay was used to evaluate the cytotoxicity of microglial cells. EtBr/EthD2 with different molecular weights (394 Da/1 293 Da) was used to measure the size of plasma membrane pores. ELISA was used to measure the level of the inflammatory factor interleukin-1β (IL-1β) in culture supernatant.@*RESULTS@#After bilirubin stimulation, the viability of microglial cells decreased and LDH release increased, both in a time-dependent manner. Compared with the control group, the bilirubin group had a significantly higher positive rate of small-molecule EtBr passing through the cell membrane (P0.05). The expression of activated Caspase-1 significantly increased at 0.5 hour after bilirubin stimulation (P<0.05), and that of activated GSDMD significantly increased at 6 hours after bilirubin stimulation (P<0.05). The release of IL-1β significantly increased at 6 hours after bilirubin stimulation and reached the peak at 24 hours (P<0.001). Compared with the bilirubin group, the VX-765+bilirubin group had a significant increase in cell viability (P<0.05) and significant reductions in the expression of activated GSDMD, the pass rate of EtBr, and the release of LDH and IL-1β (P<0.05).@*CONCLUSIONS@#Pyroptosis is involved in bilirubin-induced injury of primary cultured microglial cells.

Determination of six constituents in Qingkailing Freeze-Dried Powder for Injection by HPLC / 中成药

AIM To establish an HPLC method for the content determination of six constituents in Qingkailing Freeze-Dried Powder for Injection (cholic acid,hyodeoxycholic acid,Bubali Cornu,etc.).METHODS The content determination of adenosine,chlorogenic acid and gardenoside was performed on a 30 ℃ thermostatic XBridge C1s column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of acetonitrile-water (containing 0.1％ formic acid) flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 254 nm.The content determination of baicalin,hyodeoxycholic acid and cholic acid was performed on a 35 ℃ thermostatic XBridge C1s column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of methanol-water (containing 0.1％ formic acid) flowing at 1.0 mL/min in a gradient elution manner.RESULTS Six constituents showed good linear relationships within the ranges of 2.244-56.108,2.658-66.445,4.347-108.682,122.01-1 016.75,131.94-1 099.50,152.22-1 268.50 μg/mL (r ＞ 0.999 0),whose average recoveries (RSDs) were 101.1％ (0.46％),98.0％ (1.74％),99.7％ (0.15％),100.9％ (1.31％),98.1％(0.18％),98.2％ (1.61％),respectively.CONCLUSION This stable and reproducible method can be used for the quality control of Qingkailing Freeze-Dried Powder for Injection.

Bisphenol A alters glucose metabolism in rat Sertoli cells in vitro / 中华男科学杂志

<p><b>OBJECTIVE</b>To observe the influence of different concentrations of bisphenol A (BPA) on glucose metabolism and lactate dehydrogenase (LDH) expression in rat Sertoli cells in vitro and investigate the mechanisms of BPA inducing male infertility.</p><p><b>METHODS</b>Using two-step enzyme digestion, we isolated Sertoli cells from male Wistar rats and constructed a primary Sertoli cell system, followed by immunohistochemical FasL staining. We randomly divided the Sertoli cells into a control group to be cultured in the serum-free minimal essential medium (MEM) plus dimethyl sulfoxide (DMSO) and three experimental groups to be treated with 100 nmol/L, 10 μmol/L, and 1 mmol/L BPA, respectively, in the MEM plus DMSO. After 48 hours of treatment, we measured the proliferation of the cells by CCK-8 assay, determined the concentrations of metabolites by NMR spectroscopy, and detected the expression of LDH in the Sertoli cells by RT-PCR and Western blot.</p><p><b>RESULTS</b>The purity of the isolated Sertoli cells was (96.05 ± 1.28)% (n = 10). Compared with the control group, the 100 nmol/L, 10 μmol/L, and 1 mmol/L BPA groups showed no remarkable changes in the proliferation of Sertoli cells ([98 ± 8]%, [96 ± 3]%, and [95 ± 3]%, P >0.05), but the 10 μmol/L and 1 mmol/L of BPA groups exhibited significantly decreased concentrations of intracellular glucose ([3.89 ± 0.07] vs [3.36 ± 0.24] and [3.04 ± 0.21] pmol/cell, P <0.05) and lactate ([0.43 ± 0.06] vs [0.29 ± 0.05] and [0.20 ± 0.03] pmol/cell, P <0.05). The expression of LDH mRNA was decreased with the increased concentration of BPA, while that of LDH protein reduced only in the 1 mmol/L BPA group (P <0.05).</p><p><b>CONCLUSION</b>High-concentration BPA decreases the expression of LDH and alters glucose metabolism in Sertoli cells, and therefore may reduce the provision of lactate for germ cells and impair spermatogenesis.</p>

Contrast-enhanced ultrasonography for detecting testicular perfusion in acute testis contusion in rabbits / 中华男科学杂志

<p><b>OBJECTIVE</b>To evaluate contrast-enhanced ultrasonography (CEUS) in detecting testicular perfusion in acute testis contusion.</p><p><b>METHODS</b>We established the model of testis contusion in 11 healthy male New Zealand rabbits by randomly hitting one side of the scrotum under general anesthesia. We examined the bilateral scrotums of all the animals before, immediately after and at 2, 4 and 6 hours after modeling by color Doppler flow imaging (CDFI) and CEUS, and analyzed the time-intensity curve (TIC), arriving time (AT), time to peak intensity (TTP), peak intensity (PI), half time of descending peak intensity (HT) and area under the curve (AUC) in the healthy and injured testis, respectively.</p><p><b>RESULTS</b>CEUS exhibited a higher sensitivity in detecting tissue perfusion than CDFI. The mode of contrast agent perfusion in testicular contusion was fast in and slow out. There were no evident differences between the contused and the healthy testis in AT, TTP and PI before modeling. The contused testis showed significantly earlier AT and TTP, higher PI and larger AUC (P < 0.05) than the healthy one at different time points after modeling, but no statistically significant difference was found in HT (P > 0.05).</p><p><b>CONCLUSION</b>Accurate parameters of testicular perfusion in acute testis contusion can be quantitatively obtained by CEUS, which are of important value for the diagnosis of testis contusion.</p>

The evaluation of left ventricular strain using speckle tracking echocardiography in normal young adults：comparison of three-dimensional and two-dimensional approaches / 中华医学超声杂志（电子版）

Objective To investigate the value of three-dimensional speckle tracking imaging (3D-STI) in assessment of left ventricular (LV) strains. Methods Thirty healthy young adults examined by two-dimensional speckle tracking imaging (2D-STI) and 3D-STI. And the results of LV measurements were compared, which included mean peak systolic longitudinal strains, radial strains and circumferential strains. Also, the time consumption of these two methods was compared. Results The time needed for 3D-STI in acquisition and analysis of the images were (309.3±23.4)s, (305.5±11.2)s, while the time for 2D-STI were (490.6±14.4)s, (1261.4±39.9)s. The differences were signiifcant(t=-21.81, 69.94, both P<0.01). The global mean peak systolic radial strains was (48.59±7.68)%by 3D-STI and (33.25±7.27)%by 2D-STI. The difference was signiifcant(t=9.16, P<0.01). The global mean peak systolic longitudinal and circumferential strains were (-17.66±3.14)%, (-17.13±2.29)% by 3D-STI and (-21.35±2.46)%, (-21.97±3.84)% by 2D-STI. The differences were signiifcant(t=5.33, 5.99, both P < 0.01). The 3D-STI strains were different at different levels of LV. The longitudinal, circumferential and radial 3D-STI strains were largest at middle levels. However, 2D-STI strains didn′ t show such trend. Peak strains measured by 3D-STI and 2D-STI showed high inter-observer and intra-observer agreement in Bland-Altman chart. Conclusion 3D-STI is a novel, convenient and reproducible method to evaluate the strains of LV.

Propofol protecting rats from paraquat induced lung injury / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the protective efficacy of propofol against paraquat induced lung injury.</p><p><b>METHODS</b>One hundred and twenty-eight male Wistar rats were randomizedly divided into three groups: the control group (n = 8), the intoxication group (n = 60) and the propofol group (n = 60). One hundred and twenty rats were once administered with 5 mg/kg paraquat (PQ) by the intragastrical injection to establish the model of PQ induced lung injury. The propofol of 10 mg/kg was administered intraperitoneally in the propofol group (60 rats) twice a day for four consecutive days one hour after the rats were intoxicated while the normal saline of the same amount as propofol in the propofol group was administered in the intoxication group (60 rats) one hour after the rats were intoxicated. The intragastrical injection of 1 mg/kg normal saline was administered once in the control group (8 rats). On the first, the third, the seventh, the 14th and the 28th day after treating, the level of malondialdehyde (MDA) in plasma, bronchoalveolar lavage fluid (BALF) and lung homogenate, and the content of hydroxyproline (HPY) in lung homogenate, the cell count of BALF were detected. Meanwhile, pathological changes of lung were examined under optical microscope.</p><p><b>RESULTS</b>The level of MDA in plasma on the first, the third and the seventh day and in BALF and lung homogenate on the first and the third day in the propofol group [in plasma: (4.31 +/- 0.94), (4.04 +/- 0.87) and (3.24 +/- 1.14) nmol/ml; in BALF: (3.47 +/- 1.09) and (2.79 +/- 1.11) nmol/ml; in lung homogenate: (7.54 +/- 0.63) and (8.41 +/- 1.23) nmol/ml] were significantly lower than those in the intoxication group [in plasma: (10.15 +/- 3.15), (6.97 +/- 1.6 5) and (5.44 +/- 0.66) nmol/ml; in BALF: (5.58 +/- 1.19) and (4.86 +/- 1.89) nmol/ml; in lung homogenate: (10.20 +/- 2.43) and (10.71 +/- 171) nmol/ml, P < 0.05 or P < 0.01]. The total cell count of BALF on the first, the third and the seventh day after intoxication in the propofol group was significantly less than that in the intoxication group respectively (P < 0.05). The histological changes such as alveolar edema, hemorrhage and inflammatory cell infiltration in the propofol group were less than those in the PQ group.</p><p><b>CONCLUSION</b>Propofol could reduce the level of MDA and relieve paraquat induced lung injury.</p>