Objective To investigate the effect of long non-coding RNA growth arrest-specific transcript 5(lncRNA GAS5)on lung tissue cell apoptosis in acute lung injury(ALI)rats via regulation of the miR-452-5p/myeloid cell leukemia sequence 1(Mcl-1)pathway.Methods SD rats were randomly separated into 6 groups(10 in each group):control group,model group,pUC57 lncRNA GAS5 group,miR-452-5p antagonist group,pUC57 NC+miR-452-5p NC group,and pUC57 lncRNA GAS5+miR-452-5p agonist group.The ALI model was constructed via the intraperitoneal injection of lipopolysaccharide in the model group and drug intervention group,whereas the control group rats were intraperitoneally injected with an equal dose of physio-logical saline.At the same time,the intervention was performed,and the lung function of the rats was subsequently meas-ured.The forced vital capacity(FVC),forced expiratory volume in 0.3 second(FEV0.3)/FVC,and tidal volume(VT)were com-pared among the groups.HE staining was used to observe pathological damage in the lung tissue,and the degree of damage was evaluated via the Holfbauer score.An enzyme-linked immunosorbent assay(ELISA)was used to measure the levels of bronchoal-veolar lavage fluid(BALF)and the serum inflammatory factors interleukin-18 and IL-1β in the rats.Immunoblotting was used to detect the expression of pyroptosis-related indicator proteins[(GSDMD),nucleotide binding oligomeric domain-like receptor protein 3(NLRP3),cysteine-containing aspartate-specific protease(Caspase-1),and Caspase-11]and Mcl-1 in rat lung tissue.Real-time fluorescence quantitative PCR was used to detect the expression of lncRNA GAS5 and miR-452-5p in rat lung tissue.A dual-luciferase reporter gene experiment was performed to validate the targeted regulation of miR-452-5p by lncRNA GAS5 in rat alveolar epithelial cell L2.Results Compared with those in the control group,the FVC,FEV0.3/FVC,expression of lncRNA GAS5,expression of Mcl-1 protein and mRNA in the model group decreased(P＜0.05),and the VT,Holfbauer score,IL-18 and IL-1β levels,GSDMD,NLRP3,Caspase-1 and Caspase-11 protein expression,and miR-452-5p expression were elevated(all P＜0.05).Compared with those in the model group,the FVC,FEV0.3/FVC,expression of Mcl-1 protein and mR-NA in the pUC57 lncRNA GAS5 group and the miR-452-5p antagonist group were increased(all P＜0.05),and the VT,Holf-bauer score,IL-18 and IL-1β levels,GSDMD,NLRP3,Caspase-1 and Caspase-11 protein expression,and miR-452-5p expression were reduced(all P＜0.05).There was no obvious difference in various indicators in the pUC57-NC+miR-452-5p-NC group(all P＞0.05).Compared with those in the pUC57 lncRNA GAS5 group,the FVC,FEV0.3/FVC,expression of Mcl-1 protein and mRNA in the pUC57 lncRNA GAS5+miR-452-5p agonist group decreased(all P＜0.05),and the VT,Holfbauer score,IL-18 and IL-1β levels,GSDMD,NLRP3,Caspase-1 and Caspase-11 protein expression,and miR-452-5p expression were elevated(all P＜0.05).Conclusion lncRNA GAS5 can promote Mcl-1 expression by down-regulating miR-452-5p,thereby inhibiting the in-flammatory response and lung tissue cell pyroptosis in ALI rats,ultimately reducing lung injury and improving lung function.

In order to explore the role of pre-basic course teaching in the standardized training of residents in clinical pathology, we have independently designed and constructed a pre-basic course teaching system mainly focusing on anatomy and histoembryology, consisting of two levels of theoretical teaching (small lectures by students and systematic lectures by instructors) and two dimensions of practical training (sample collection teaching and case teaching). This teaching model centering on participatory lecturing, practice, summarization, assessment, and feedback has been demonstrated effective. The results showed that the theoretical and practical assessment scores of the experimental group [(458.80±17.60) and (415.40±19.30), respectively] were significantly higher than those of the control group [(444.50±20.90) and (398.80±23.70), respectively]. Among 28 students of grades 2019 to 2021 surveyed for teaching effectiveness, 96.43% were satisfied with the teaching model. The established teaching model provides new ideas for the reform of teaching methods in the standardized training of residents in clinical pathology.

This study aimed to explore the effect of miR-23b-3p on the differentiation of goat intramuscular preadipocytes, and to confirm whether miR-23b-3p plays its roles via targeting the PDE4B gene. Based on the pre-transcriptome sequencing data obtained previously, the miR-23b-3p, which was differentially expressed in goat intramuscular adipocytes before and after differentiation, was used as an entry point. real-time quantitative-polymerase chain reaction (qPCR) was used to detect the expression pattern of miR-23b-3p during the differentiation of goat intramuscular preadipocytes. The effects of miR-23b-3p on adipose differentiation and adipose differentiation marker genes were determined at the morphological and molecular levels. The downstream target genes of miR-23b-3p were determined using bioinformatics prediction as well as dual luciferase reporter assay to clarify the targeting relationship between miR-23b-3p and the predicted target genes. The results indicated that overexpression of miR-23b-3p reduced lipid droplet accumulation in goat intramuscular adipocytes, significantly down-regulated the expression levels of adipogenic marker genes AP2, C/EBPα, FASN, and LPL (P < 0.01). In addition, the expressions of C/EBPβ, DGAT2, GLUT4 and PPARγ were significantly downregulated (P < 0.05). After interfering with the expression of miR-23b-3p, lipid droplet accumulation was increased in goat intramuscular adipocytes. The expression levels of ACC, ATGL, AP2, DGAT2, GLUT4, FASN and SREBP1 were extremely significantly up-regulated (P < 0.01), and the expression levels of C/EBPβ, LPL and PPARγ were significantly up-regulated (P < 0.05). It was predicted that PDE4B might be a target gene of miR-23b-3p. The mRNA expression level of PDE4B was significantly decreased after overexpression of miR-23b-3p (P < 0.01), and the interference with miR-23b-3p significantly increased the mRNA level of PDE4B (P < 0.05). The dual luciferase reporter assay indicated that miR-23b-3p had a targeting relationship with PDE4B gene. MiR-23b-3p regulates the differentiation of goat intramuscular preadipocytes by targeting the PDE4B gene.
Animals ; MicroRNAs/metabolism* ; Goats/genetics* ; PPAR gamma/metabolism* ; Adipogenesis/genetics* ; Cell Differentiation/genetics* ; Luciferases ; RNA, Messenger

Objective To explore the expression of urothelial carcinoma-associated 1 ( UCA1 ) in pancreatic cancer cell lines and its influence on the invasion and metastasis of the pancreatic cancer cells .Methods The expression of UCA1 in pancreatic cancer tissues and paired adjacent normal tissues ( 11 cases ) and 5 pancreatic cancer cell lines was analyzed by real-time PCR.The level of UCA1 in BxPC-3 was knocked down by small interfering RNA . The ability of invasion and migration in vitro of transfected BxPC-3 was detected by Transwell invasion assay and wound healing assay .The protein levels of MMP-2 and MMP-9 were measured by Western blot experiment .Results The expression level of UCA1 in pancreatic cancer tissues was higher than that in paired adjacent normal tissues , and UCA1 differentially expressed in 5 pancreatic cancer cell lines .Down-regulation of UCA1 by siRNA suppressed the expression of MMP-2 and MMP-9 in BxPC3, and dramatically impaired the ability of invasion and migration of BxPC-3.Conclusions UCA1 is over-expressed in pancreatic cancer , and down-regulation of UCA1 attenuates the capacity of invasion and metastasis in vitro of BxPC-3 by decreasing MMP-2 and MMP-9.

Objective To investigate the effect of miR-200 a mimic on transforming growth factor β1-mediated acti-vation and collagen secretion of rat pancreatic stellate cells .Methods PSCs were isolated and cultured from pan-creatic tissue and identified by desmin , GFAP and α-SMA immunofluorescence staining .PSCs of 2nd generation were divided into control group , TGF-β1 group, TGF-β1+miR-NC group and TGF-β1+miR-200a mimic group.α-SMA and collagen Ⅰ protein were measured by Western blot and immunofluorescence staining .The mRNA ofα-SMA and collagen Ⅰ and the expression of miR-200a were detected by quantitative real-time PCR.Results TGF-β1 stimulates the activation of PSCs and promote collagen synthesis in time-dependment manner ( P<0.05 ) . After transfection of the mimic , treating with the same concentration of TGF-β1, the expressions of protein and mR-NA of both α-SMA and collagen Ⅰ decreases significantly ( P<0.01 ) .Conclusions Over-expression of miR-200 a significantly attenuates α-SMA activity and further affects the collagen synthesis of TGF-β1-dependent activa-tion of PSCs.The mechanisms are potentially related to the biological effects of TGF-β1.

OBJECTIVETo investigate the reversal effect of targeted modulation of bcl-2 expression by miR-15a and miR-16 on drug resistance of human colon cancer cells.

METHODSMimics or inhibitors of miR-15a and miR-16 were transfected into HCT8 or HCT8/VCR cells with the help of Lipofectamine 2000. The expressions of miR-15a and miR-16 mRNA were detected by RT-qPCR. The levels of bcl-2 and P-gp proteins were measured by Western blot. The inhibitory effects of VCR on growth of HCT8 and HCT8/VCR cells were detected by CCK8.

RESULTSAfter transfection of the mimics, the expression of miR-15a in the blank control group, negative control group and miR-15a mimic group was 1.00, 0.87 ± 0.24, and 223.44 ± 59.07, respectively, and miR-15a was increased significantly (P < 0.001). The expression of miR-16 in the blank control group, negative control group and miR-16 mimic group was 1.00, 0.66 ± 0.19, and 107.32 ± 22.58, respectively, and miR-16 expression was increased significantly (P < 0.001). The Western blot assay showed that the relative expressions of bcl-2 protein in the blank control group, negative control group, miR-15a mimic group and miR-16 mimic group were 1.00, 0.97 ± 0.02, 0.51 ± 0.06, and 0.65 ± 0.03, respectively, and the expression of bcl-2 protein was decreased significantly (P < 0.05), however, the expressions of P-gp protein showed no significant difference. The CCK8 test showed that at 1, 5, 25 and 125 µg/ml concentration of VCR, the survival rates of HCT8/VCR cells were basically the same in the blank control group, negative control group, miR-15a mimic group and miR-16 mimic group, but the survival rate of HCT8/VCR cells was significantly decreased after transfection of mimics (P < 0.05). After transfection of the inhibitors, the expressions of both miR-15a and miR-16 were decreased significantly (P < 0.001). The Western blot showed that the expression of bcl-2 protein was increased (P < 0.05), while the expression of P-gp protein showed no significant difference. The CCK8 test showed that the survival rate of HCT8 cells which were transfected with inhibitors was significantly higher than that of the blank control group (P < 0.05).

CONCLUSIONSmiR-15a and miR-16 may reverse the drug resistance in human colon cancer cells. A possible mechanism is regulating the expression of bcl-2.

ATP-Binding Cassette, Sub-Family B, Member 1 ; Apoptosis ; Apoptosis Regulatory Proteins ; metabolism ; Cell Line, Tumor ; Colonic Neoplasms ; Drug Resistance ; Humans ; MicroRNAs ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; RNA, Messenger ; Transfection

Objective To investigate the effect of sodium valproate in suppression of cell proliferation and arrest of cell cycle of in human hepatoma cell line SMMOL/LC-7721 and pancreatic cancer cell line PaTu8988.Methods Hepatoma cell line SMMOL/LC-7721 and pancreatic cancer cell line PaTu8988 were inoculated on the culture plate,cultured in the DMEM medium,they were intervened with sodium valproate in concentration of 0.2 mmol/L (0.2 mmol/L group),1.0 mmol/L (1.0 mmol/L group),5.0 mmol/L (5.0 mmol/L group) and dimethyl sulfoxide (control group) for 48 h respectively.Absorbance was measured by enzymelinked immunosorbentassay equipment,and inhibition ratio was calculated.Cell cycle was detected by flow cytometry.Results The absorbance of hepatoma cell line SMMOL/LC-7721 and pancreatic cancer cell line PaTu8988 in 5.0 mmol/L group were significantly lower than those in control group and 0.2 mmol/L group (0.569 ±0.059 vs.0.706 ±0.033 and 0.760 ±0.020,2.068 ±0.178 vs.2.793 ±0.144 and 2.663 ± 0.078,P ＜ 0.05),the absorbance of pancreatic cancer cell line PaTu8988 in 1.0 mmol/L group (2.432 ± 0.084) was significantly lower than that in control group and 0.2 mmol/L group (P ＜ 0.05).With the sodium valproate concentration increased,inhibition rate of tumor cell increased gradually,the inhibition rate of hepatoma cell line SMMOL/LC-7721 and pancreatic cancer cell line PaTu8988 in 5.0 mmol/L group was 23.5％ and 25.9％ respectively.Compared with control group,with the sodium valproate concentration increased in 0.2,1.0,5.0 mmol/L group,the proportion of G1 phase cell increased gradually in hepatoma cell line SMMOL/LC-7721 [(49.25 ± 1.63)％,(65.26 ± 2.34)％,(83.13 ± 1.78)％ vs.(49.22 ± 4.35)％],the proportion of S phase cell decreased gradually [(26.84 ± 2.30)％,(17.76 ± 3.90)％,(3.38 ± 0.65)％ vs.(29.21 ± 2.35)％],cell cycle showed G1 phase arrest,there was significant difference (P ＜ 0.05).Compared with control group and 0.2 mmol/L group,the proportion of G2 phase cell increased in pancreatic cancer cell line PaTu8988 in 1.0 and 5.0 mmol/L group [(26.80 ± 1.50)％,(36.58 ± 3.78)％ vs.(12.00 ± 4.62)％,(16.54 ± 2.26)％],cell cycleshowed G2 phase arrest,there was significant difference (P ＜ 0.05).Conclusion Sodium valproate mightsignificantly suppress the cell proliferation in hepatoma cell line SMMOL/LC-7721 and pancreatic cancercell line PaTu8988 and induce cell cycle arrest,it is clinically promising antitumor drug.

Objective To investigate the correlation between the ratio change of circulating myeloid-derived suppressor cells (MDSCs) and cellular immune function in healthy volunteers,chronic gastritis patients,gastric intraepithelial neoplasia patients and gastric cancer patients.Methods From February 2011 to July 2011,129 peripheral blood samples were collected,including 32 healthy volunteers,48 chronic gastritis patients,27 gastric intraepithelial neoplasia patients and 22 gastric cancer patients.The percentages of peripheral blood MDSC,T lymphocyte subsets and regulatory T cells (Treg) were determined by flow cytometry.The data were analyzed by one way analysis of variance,pearson and spearman correlation.Results The percentages of circulating MDSC,CD8+ T lymphocyte and Treg were highest in gastric cancer patients (9.63％±3.24％,10.03％ ± 1.26％,69.45％±3.42％) and lowest in healthy volunteers (0.92％±0.33％,4.12％ ±0.99％,32.35％ ±4.83％).Those of gastric intraepithelial neoplasia patients (5.13％ ± 1.30％,7.54％ ± 0.79％,53.26％±4.30％) were lower than gastric cancer patients but higher than chronic gastritis patients (2.76％ ±0.64％,6.28％ ±0.61％,42.37％ ±4.02％).The differences among each groups were statistically significant (F=24.85,20.88,37.84,all P＜0.05).However,the percentage of circulating CD4+T lymphocyte was highest in healthy volunteers (65.10％±4.10％),55.15％ ± 4.00％ in chronic gastritis group,42.23％ ± 3.91％ in gastric intraepithelial neoplasia group,and lowest in gastric cancer group (26.84％ ± 3.69％).The differences among each groups were statistically significant (F=46.80,P＜0.05).A significant correlation between circulating MDSC and TNM stages of gastric cancer was also observed (r=0.856,P＜0.01).The percentage of circulating MDSC was positively correlated with Treg percentage (r =0.862,P ＜ 0.01),and negatively correlated with CD4+/CD8+ ratio (r=-0.768,P＜0.01).Conclusion The increase of MDSC percentage in peripheral blood is correlated with human cellular immune function,which might play an important role in the tumor immune evasion during the development of gastric cancer.

Objective To investigate the expression of DNMT3b and its correlation with clinicopathological parameters of pancreatic cancer.Methods The expressions of DNMT3b protein in 12 pancreatic cancer tissues and para-cancerous tissues were detected by Western blot.The expressions of DNMT3b protein in 59 pancreatic cancer tissues were detected by immunohistochemistry.The association of DNMT3b expression and clinicopathological parameters of pancreatic carcinoma were analyzed.Results Western blot results showed the expressions of DNMT3b protein in pancreatic cancer tissues and para-cancerous tissues were 0.69 ±0.13and 0.14 ±0.03,and the protein level of DNMT3b in pancreatic cancer tissues were significantly higher than that in para-cancerous tissues (t =4.464,P ＜0.05 ).Immunohistochemistry examination showed that the positive rates of DNMT3b protein were 59％ in pancreatic cancer tissues and negative in para-cancerous tissues.DNMT3b expression was positively correlated with the TNM stage and lymph node metastasis.Conclusions DNMT3b is highly expressed in pancreatic cancer tissues,and it is related with malignant biological behaviors of pancreatic cancer cells.

Objective To evaluate operative methods and the clinical effect for comminuted calcaneal fractures.Methods 21 cases(24 feet)of comminuted calcaneal fractures were treated by one stage bone grafting plus open reduction and internal fixation with plastic calcaneus titanium plate.The fractures were classified according to the Sanders system into Sanders Ⅱ,Ⅲ and Ⅳ subgroups.Results All cases were followed up for 9～20 months(14months in average).All cases recovered with good healing of fracture.According to Maryland score,the results were excellent in 14 feet,good in 7 feet,fair in 2 feet,and poor in 1 foot.The excellent and good rate was 87.5％.Conclusions By using the method of internal fixation with plastic calcaneus titanium plate,fracture site can be well exposed,which is helpful to recover calcaneus to its normal length,width,height,Gissane angle and Bohler angle.A reliable internal fixation is helpful for early stage functional excise after surgery,which can maximize the recovery of the ankle function.