Construction and performance evaluation of ultra-fast thermal cycle fluorescence quantitative PCR system / 医疗卫生装备

Objective To design an ultra-fast thermal cycle fluorescent quantitative PCR system for on-site detection of pathogenic nucleic acids and evaluate its performances.Methods An ultra-fast thermal cycle fluorescent quantitative PCR system was developed with the components of a flat reaction cup,an ultra-fast thermal cycle module,a fluorescence detection module with fixed optical path and a data processing module based on the smartphone platform.The ultra-fast thermal cycle module was composed of a heating unit and a cooling unit,of which the heating unit was made of ceramic sheet and Ag/Pb alloy and the cooling unit consisted of a high-speed magnetic levitation cooling fan and a double-curved throat;the fluorescence detection module with fixed optical path was prepared with injection molding process,and made up of a light source excitation unit and a light detector unit;the data processing module based on the smartphone platform included a Bluetooth serial port adapter unit and a smartphone App,which used C2540F256 chip from TI company for developing the Bluetooth serial port adapter and Android Studio for the App.The ultra-fast thermal cycle fluorescent quantitative PCR system was used to detect influenza A/B virus and SARS-CoV-2 to verify its performances.Results The ultra-fast thermal cycle fluorescent quantitative PCR system realized rapid nucleic acid detection of influenza A/B virus and SARS-CoV-2,and the detection results were in high agreement with those by conventional real-time quantitative PCR.Conclusion The ultra-fast thermal cycle fluorescence quantitative PCR system gains advantages in small size and light weight,and can be used for rapid on-site detection of pathogen nucleic acids.[Chinese Medical Equipment Journal,2023,44(11):15-20]

Stercoral colonic diverticulum perforation with jejunal diverticulitis mimicking upper gastrointestinal perforation / 中华医学杂志(英文版)

Stercoral perforation of the colon is an unusual pathological condition with fewer than 150 cases reported in the literature to date. We present a case of stercoral colonic perforation mimicking upper gastrointestinal perforation, which was diagnosed by computed tomography preoperatively. However, at laparotomy, stercoral colonic diverticulum perforation with jejunal diverticulitis became the most appropriate diagnosis.

Effect of protein kinase C/transforming growth factor beta 1 pathway on activation of hepatic stellate cells / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the effect of protein kinase C (PKC)/transforming growth factor beta 1 (TGF beta1) pathway on activation of hepatic stellate cells (HSC).</p><p><b>METHODS</b>HSC rHSC-99 cell line was used in three groups in this study. Group A served as a control. In group B the HSC were incubated with PKC agonist PMA (0.5 micromol/L), and in group C the cells were incubated with PKC inhibitor calphostin C (100 nmol/L). The PKC activities were detected at different incubation time points (0, 3, 6, 12 and 24 h). Western blot and RT-PCR were used to detect the expression of TGF beta1, Smad 4, collagen type I, III and alpha-smooth muscle actin (alpha-SMA) at the 24 h point. Cell proliferation was assessed by MTT colorimetric assay.</p><p><b>RESULTS</b>PMA increased the activity of PKC significantly, whereas calphostin C inhibited the activity of PKC. The increased activity of PKC promoted the HSC to express TGF beta1, Smad 4, collagen type I, III and alpha-SMA. In comparison with the controls, the expressions of TGF beta1, Smad 4, collagen type I, III and alpha-SMA increased 4.8, 13.1, 2.4, 1.8 and 1.3 fold respectively (P < 0.01). PKC promoted the proliferation of HSC. The above effects were inhibited by the inhibition of PKC activity.</p><p><b>CONCLUSION</b>Changing of PKC activity can regulate and control the expression of TGF beta1, which may play a role in regulating the activation of HSC.</p>

Downregulation of transforming growth factor-beta1 and platelet-derived growth factor gene expression by interleukin-10 in murine hepatic stellate cells in response to experimental liver injury / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the effect of interleukin-10 (IL-10) on the expression of transforming growth factor-beta(1) (TGFbeta(1)) and platelet-derived growth factor (PDGF) in hepatic stellate cells (HSC) during liver injury.</p><p><b>METHODS</b>The adenovirus vector (the titer was 1 x 10(7) efu/ml) encoded IL-10 gene was used to transfect the rat via the vein of caudal. At the same time, CCl(4) was injected into rat by a hypodermic injection. These processes went on twice a week. After eight weeks, the liver were perfused with collagenase IV and purified by density gradient centrifugation with Nycodenz for separate HSC. The level of IL-10 was measured by ELISA method; The expression of PDGF and TGFbeta(1) in HSC was detected by semi-quantitative RT-PCR and Western-blot methods.</p><p><b>RESULTS</b>The level of IL-10 in therapy group (adenovirus vector encoding IL-10 gene group) was higher than that in non-therapy group (adenovirus vector without IL-10 gene and PBS group); The expression of TGFbeta(1) mRNA, TGFbeta(1) protein and PDGF mRNA, PDGF protein in therapy group were significantly lower than that in non-therapy group (P < 0.05).</p><p><b>CONCLUSION</b>Downregulating the TGFbeta(1) and PDGF expression could be the passageway by which IL-10 alleviate the degree of proliferation and activation in hepatic stellate cells.</p>

An in vivo study on interlukin-10 inhibiting the expression and secretion of collagen I and IV in hepatic stellate cells / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To investigate the effect of interlukin-10 (IL-10) on expression and secretion of collagen I, IV in rat's hepatic stellate cells (HSC) of livers injured by CCl4.</p><p><b>METHOD</b>The adenovirus vector encoded IL-10 gene was used to transfect rats with liver injury via the caudal veins. HSC were isolated and purified from the rat livers by collagenase IV perfusion and density gradient centrifugation with Nycodenz. The expression of collagen I, IV mRNA in HSC was detected by semi-quantitative RT-PCR method and the secretion of collagen I, IV in culture serum of HSC by ELISA method. The quantity of collagen was measured in the van Gieson stained histological liver preparations.</p><p><b>RESULTS</b>The expression and secretion of collagen I, IV in the adenovirus vector encoding IL-10 gene group were significantly lower than those in the adenovirus vector without IL-10 gene group and the control group (P < 0.05). The quantity of collagen in the treatment group was lower than that in the control group.</p><p><b>CONCLUSION</b>IL-10 can inhibit collagen I, IV expression and secretion in rat HSC.</p>