Expression of glucose transporter 3 in cutaneous squamous cell carcinoma and its effect on the proliferation, invasion and migration of A431 cells / 中华皮肤科杂志

Objective:To determine the expression of glucose transporter 3 (GLUT3) in cutaneous squamous cell carcinoma (cSCC), and to evaluate its effect on the cSCC cell line A431.Methods:From June 2016 to December 2020, 22 paraffin-embedded tissue specimens were collected from patients with pathologically confirmed cSCC in the Department of Dermatology, Peking University Third Hospital, and 20 discarded normal skin tissues after dermatological surgeries served as controls. Immunohistochemical assay was performed to determine the GLUT3 expression in cSCC tissues and normal skin tissues. Cultured A431 cells were divided into two groups: GLUT3 overexpression group transfected with a lentiviral vector carrying the SLC2A3 gene, and negative control group transfected with an empty lentiviral vector. Real-time fluorescence-based quantitative PCR and Western blot analysis were conducted to determine the mRNA and protein expression of GLUT3 in A431 cells in different groups, the cell proliferation assay (MTS assay) was performed to estimate the cell proliferative activity, and the live-cell analysis system Incucyte S3 was used for real-time evaluation of the migratory and invasive abilities of A431 cells in different groups. The relative glucose consumption and lactic acid production in A431 cells at 48 hours were measured by using glucose and lactic acid assay kits, retrospectively. Two independent samples t-test was used for comparisons between two groups, and one-way analysis of variance was used for comparisons among multiple groups. Results:The GLUT3 expression was significantly higher in the cSCC tissues than in the normal skin tissues (immunohistochemical assay score: 9.39 ± 2.56 points vs. 2.30 ± 2.60 points; t = 8.91, P < 0.05). Compared with the negative control group, the mRNA and protein expression of GLUT3 markedly increased in the GLUT3 overexpression group. MTS assay showed significantly increased proliferative activity of A431 cells in the GLUT3 overexpression group compared with the negative control group after 24- and 96-hour treatment ( t = 2.49, 3.54, P = 0.048, 0.012, respectively); cell fusion rates in the scratched area were significantly higher in the GLUT3 overexpression group than in the negative control group in the cell migration assay at 6, 12 18, and 24 hours and cell invasion assay at 12, 18, and 24 hours (all P < 0.05). At 48 hours, the relative glucose consumption and lactic acid production in A431 cells were significantly higher in the GLUT3 overexpression group than in the negative control group ( t = 2.98, 2.20, P = 0.011, 0.038, respectively) . Conclusion:GLUT3 was highly expressed in the cSCC tissues, and may participate in the occurrence and development of cSCC by providing energy to cSCC cells via promoting glucose uptake in cells to enhance their proliferative, migratory and invasive abilities.

Design and application of an anti-pull device for medical drainage tube / 中华危重病急救医学

The management of drainage tube is an important part of nursing work. Patient restraint and tube fixation cannot effectively prevent unplanned extubation (UEX) when the tube is accidentally pulled by violence. The nursing innovation team of Henan Provincial People's Hospital designed a medical drainage tube anti-pull device in order to change the existing technology of preventing drainage tube disconnecting by means of restraint and fixation, and to interfere with the basic cause of drainage tube disconnection, and obtained the national utility model patent (patent number: ZL 2020 2 2843025.1). The design of sleeve and clasp is that when the drainage tube is pulled by accidental violence, the friction fastener clamps the drainage tube mechanically to achieve the purpose of braking the drainage tube and prevent the drainage tube from coming out. Card sleeve ring fracture design can be applied to drainage tubes of different diameters, and the buzzer device at the instant of the snap ring into the card set warning medical staff to the occurrence of risk events, so that the nurse can come in the first place for effective treatment, which is a fuse for surgical drainage tubes and is to timely and effectively prevent UEX.

Targeting BMI-1-mediated epithelial-mesenchymal transition to inhibit colorectal cancer liver metastasis

Liver is the most common metastatic site for colorectal cancer (CRC), there is no satisfied approach to treat CRC liver metastasis (CRCLM). Here, we investigated the role of a polycomb protein BMI-1 in CRCLM. Immunohistochemical analysis showed that BMI-1 expression in liver metastases was upregulated and associated with T4 stage, invasion depth and right-sided primary tumor. Knockdown

Therapeutic effect of human umbilical cord mesenchymal stem cells on imiquimod-induced psoriasis-like mouse models / 中华皮肤科杂志

Objective:To investigate the therapeutic effect of human umbilical cord mesenchymal stem cells (MSCs) on psoriasis-like mouse models induced by imiquimod and the underlying mechanisms.Methods:Eighteen C57BL/6 mice were randomly and equally divided into vaseline group, model group and treatment group according to a random number table. The mice in the model group and treatment group received topical treatment with 5% imiquimod cream at a dose of 62.5 mg once a day for 6 consecutive days on the shaved back, and those in the vaseline group received the treatment with the same amount of vaseline ointment; the mice in the treatment group were injected with 1.5×10 6 human umbilical cord MSCs via the caudal vein on days 1 and 4. The severity of skin lesions on the back of the mice was assessed everyday according to the psoriasis area and severity index (PASI) . Twenty-four hours after the last treatment, that is, on day 7, blood samples were taken, and the mice were sacrificed. The dorsal skin tissues were resected and subjected to hematoxylin and eosin (HE) staining. A single cell suspension of the resected spleen was prepared, and flow cytometry was performed to detect the Th1 and Th17 cell subsets in the spleen cells. Enzyme-linked immunosorbent assay was conducted to detect serum levels of cytokines interleukin (IL) -17A and tumor necrosis factor (TNF) -α. One-way analysis of variance was used for comparisons among groups, Tukey test for multiple comparisons, and repeated measures analysis of variance for the analysis of changes in the PASI score over time. Results:On day 7, there was obvious scaly erythema on the back of the mice in the model group, and the skin thickness and number of infiltrating inflammatory cells were significantly higher in the model group (78.73 ± 23.11 μm, 36.16 ± 2.95 cells/mm 2) than in the vaseline group (13.28 ± 4.57 μm, 13.33 ± 1.15 cells/mm 2, q=19.25, 7.21, respectively, both P < 0.001) . The treatment group showed significantly decreased PASI score, epidermal thickness and number of infiltrating inflammatory cells compared with the model group (all P < 0.001) . The percentage of Th17 cell subsets in the spleen cells and serum level of TNF-α were significantly lower in the treatment group than in the model group (both P < 0.05) . There were no significant differences in the spleen weight, spleen index, spleen cell count, Th1 cell percentage or serum IL-17A level between the treatment group and the model group (all P>0.05) . Conclusion:Human umbilical cord MSCs can effectively alleviate skin inflammation induced by imiquimod in the psoriasis-like mouse models, likely by inhibiting Th17 cell formation and TNF-α expression.

Effect of chidamide combined with matrine on proliferation and apoptosis of cutaneous T-cell lymphoma cell lines HH and Hut78 and possible apoptotic mechanisms / 中华皮肤科杂志

Objective To evaluate the effect of chidamide combined with matrine on proliferation and apoptosis of cutaneous T-cell lymphoma (CTCL) cell lines HH and Hut78,and to explore their apoptotic mechanisms.Methods Both HH and Hut78 cells were treated with 0.4 μmol/L chidamide and 0.6 g/L matrine alone or in combination for 24,48 and 72 hours,with those treated with dimethyl sulfoxide (DMSO) serving as control groups.MTS assay was performed to deteet cellular proliferation rates of HH and Hut78 cells at each time point.After 48-hour treatment,flow cytometry was conducted to detect cell apoptosis,and Western blot analysis to determine expression of apoptosis-related proteins in these cells.Statistical analysis was carried out by using repeated measures analysis of variance,one-way analysis of variance,and least significant difference (LSD)-t test for multiple comparisons.Results Compared with DMSO,chidamide and matrine alone or in combination could inhibit the proliferation of HH and Hut78cells to different extents (F =15.88,558.26,P ＜ 0.05,＜ 0.001,respectively).At 48 hours,the apoptosis rate in Hut78 cells was significantly higher in the matrine group (20.98％ ± 1.53％),chidamide group (22.44％ ± 7.74％) and combination group (44.53％ ± 1.85％) than in the control group (8.42％ ± 4.23％;LSD-t =4.76,5.31,13.69 respectively,all P ＜ 0.05),as well as in the combination group than in the matrine group and chidamide group (LSD-t =8.93,8.37 respectively,both P ＜ 0.01);no significant differences were observed in the apoptosis rate of HH cells between the matrine group (13.98％ ± 3.86％)or chidamide group (13.61％ ± 1.62％) and control group (11.44％ ± 1.43％,both P ＞ 0.05),while the combination group (20.94％ ± 0.64％) showed a significantly higher apoptosis rate compared with the control group,matrine group and chidamide group (LSD-t =7.37,5.40,5.69 respectively,all P ＜ 0.05).In the case of HH cells,the combination group showed significantly higher cleaved caspase-3 expression (all P ＜ 0.05),but significantly lower protein expression of E-cadherin,nuclear factor (NF)-κB,phosphorylated-Bad (p-Bad) and Bcl-2 compared with the other 3 groups (all P ＜ 0.05).In the case of Hut78 cells,the expression of E-cadherin,NF-κB,p-Bad and Bcl-2 was significantly lower in the matrine group,chidamide group and combination group than in the control group (all P ＜ 0.05),while cleaved caspase-3 expression was significantly higher in the chidamide group and combination group than in the control group (both P ＜0.05).No matter in HH cells or in Hut78 cells,there were no significant differences in Bad protein expression between the 4 groups (all P ＞ 0.05).Conclusion Chidamide in combination with matrine can inhibit proliferation and induce apoptosis of HH and Hut78 cells,likely by regulating the expression of apoptosis-related proteins E-cadherin,NF-κB,p-Bad,Bcl-2 and cleaved caspase-3.

Preparation and Quality Control of Coenzyme Q10 Long-circulating Liposomes and Lyophilized Prepara-tion / 中国药房

OBJECTIVE:To prepare Coenzyme Q10 long-circulating liposomes,establish the determination method of content and entrapment efficiency,and prepare it into lyophilized preparation to improve its stability. METHODS:Coenzyme Q10 long-cir-culating liposomes were prepared by film dispersion method. Particle size and Zeta potential of liposomes were determined,and HPLC assay was used to determine the content of coenzyme Q10. Free drugs and liposomes were separated using protamine aggre-gation method,and the encapsulation efficiency was calculated. Lyophilized preparation was prepared by coenzyme Q10 long-circu-lating liposomes,and the changes of content and encapsulation efficiency of drugs were determined 0,30 and 90 days after lyophi-lization. RESULTS:The liposomes were homogeneous in size with mean diameter of(166.0±5.3)nm and Zeta potential of(-22.2± 1.4)mV. Average content(the percentage of content accounted for labeled amount)and entrapment efficiency of 3 batches of sam-ple were 98.2%(RSD=2.8%) and 93.2%(RSD=4.6%),respectively. Compared with 0 d after lyophilization,coenzyme Q10 long-circulating liposomes had no obvious change in the content and encapsulation efficiency 90 d after lyophilization. CONCLU-SIONS:Coenzyme Q10 long-circulating liposomes with high quality and entrapment efficiency and lyophilized preparation being stored stably for 90 d have been prepared successfully.

Effects of safflower oil on adipose-associated gene expression in mice / 中华内分泌代谢杂志

Seventy C57BL/6 mice were divided into control group,lard group,lard transferred into safflower oil group,and safflower oil group.After 20 weeks,the white adipose tissues were taken to analyse the gene differentiational expression prolife.The results showed that body weight,blood glucose and lipids,and insulin levels in lard group were higher than those in control group( all P＜0.05 ),which were decreased after lard was transferred into safflower oil for 10 weeks( all P＜0.05 ).Safflower oil regulated some of the orexigenic genes,anorectic genes,and the genes involved in energy expenditure in adipose tissues such as opioid receptor,glucagon,and PPARα.

Clinical significance of histological examination and noninvasive measurements in renal osteodystrophy / 中华肾脏病杂志

Objective To analyze the histological changes of bone diseases and to investigate the noninvasive measurements for diagnosing renal osteodystrophy (ROD) in maintenance dialysis patients . Methods Ninety-one patients were selected to receive bone biopsy . The bone samples were stained with HE, toluidine blue and Masson, and were examined with light microscopy . The levels of immunoreactive parathyroid hormone (iPTH), osteoprotegerin (OPG),sRANKL and osteocalcin (OCN) were determined in the patients enrolled from 2004 to 2006 . The level of iPTH was measured by radioimmunoassay . OPG and sRANKL were measured by ELISA,and OCN was measured by chemiluminescence . Results The incidence of ROD in the maintenance patients was 100% . According to the histological appearance, 50 cases (54 .9%) were high turnover bone disease (secondary hyperparathyroid bone disease), 9 cases (9 .9%) were low turnover bone diseases(osteomalacia and adynamic bone disease), and 32 cases(35 .2% ) were mixed bone disease . The level of iPTH in patients with ROD was significantly increased compared with healthy controls . It was the lowest in low turnover bone diseases . There was no difference among three types of ROD . OPG level was significantly increased compared with healthy controls [(2176 .58±1576 .08) pmol/L vs (1310 .46±1254 .00) pmol/L, P＜0 .05] . The level in high turnover bone diseases was higher than that of the healthy controls [(2261 .85±1712 .22) pmol/L vs (1310 .46±1254 .00) pmol/L, P＜0 .05] . There was no difference among three types of ROD .sRANKL level in high turnover bone disease was significantly increased compared with healthy controls [(0 .328±0 .524)pmol/L vs (0 .084±0 .190) pmol/L, P＜0 .05] . OCN level was also higher than that of the healthy controls (P＜0 .05), and the OCN level in low turnover ROD was the lowest among three types of ROD . OCN level in mixed ROD was dramatically increased as compared to low turnover ROD [(226 .633±66 .455) pmol/L vs (193 .03±104 .269) pmol/L, P ＜0 .05] .Conclusions The histological changes of bone disease can be indicated by iPTH level, but the types of ROD can not be distinguished according to iPTH level neither be differentiated by the levels of OPG, sRANKL and OCN . Bone histomorphometry is still the golden standard for diagnosing renal osteodystrophy .

Influence of spleen on cell cycle and expression of proliferating cell nuclear antigen of the liver during diethylnitrosamine-induced rat hepatocarcinogenesis / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the role of the spleen on hepatocyte proliferative activity during experimental hepatocarcinogenesis in rats.</p><p><b>METHODS</b>Cell-cycle and expression of proliferating cell nuclear antigen (PCNA) of hepatocytes were studied by immunohistochemical and flow cytometric technique in two groups: splenectomy (45 rats) and sham-operation with laparotomy (45 rats).</p><p><b>RESULTS</b>(1) Hepatocirrhosis was formed in group A earlier than in group B. Marked degenerative changes of the liver parachyma showing vacuolization of hepatocytes were observed in hepatic lobules. (2) The proliferative level of hepatocytes increased with the progression of hepatocarcinogenesis, and topped at the 18 th week. (3) The proliferative level of the splenectomy group was lower than that of the sham-operation group in the mid-stage of carcinogenesis (t = 4.76, P < 0.05). After stopping feeding diethylnitrosamine (DENA), however, no difference was found in the hepatocyte proliferative activity between the two groups at the 18th week.</p><p><b>CONCLUSIONS</b>There is a close relationship between hepatic proliferative activity and spleen, and the spleen may play an important role in facilitating hepatic proliferation.</p>

A study of combination therapy for hyperphosphatemia with calcium-containing phosphate binders and low-calcium dialysate in hemodialysis patients / 中国实用内科杂志

1.78 mmol/L,intact parothyroid(iPTH)2.37 mmol/L,calcium dose was 1 000 mg/d;if serum Ca~ 2+ was0.05].After low-calcium dialysis(1.25 mmol/L Ca~ 2+ dialysate)for 6 months:the serum Ca~ 2+ level had no change[(2.37?0.26)mmol/L to (2.41?0.24)mmol/L];the serum P level had decreased significantly from[(2.60?0.47)mmol/L to (2.29?0.58)mmol/L];the iPTH had increased significantly from[(130.7?84.6)pg?mL~ -1 to (169.2?105.8)pg?mL~ -1 ,P