Objective:To investigate the relationship between the expression level of long non-coding RNA transforming growth factor β2-antisense RNA1 (lncRNA TGFB2-AS1) and placental spiral artery recasting in the placenta of preeclampsia.Methods:A total of 108 pregnant women with severe preeclampsia who were hospitalized in Zaozhuang Maternal and Child Health Hospital and delivered by cesarean section from Oct. 2019 to Jun. 2021 were selected as the research objects, and they were divided into the late-onset preeclampsia group (late-onset severe preeclampsia pregnant women, 56 cases) and early-onset preeclampsia group (early-onset severe preeclampsia pregnant women, 52 cases) ; at the same time, 58 normal pregnant women were selected as the normal pregnancy group. The general data of pregnant women were collected, such as age, systolic blood pressure and diastolic blood pressure. Real-time fluorescent quantitative PCR (qRT-PCR) method was used to detect the expression level of lncRNA TGFB2-AS1 in placental tissues, a scanning electron microscope was used to measure the lumen area and wall thickness of spiral arteries. Pearson correlation analysis method was used to analyze the correlation between the level of lncRNA TGFB2-AS1 in the placenta tissue and the thickness of the spiral artery wall and the area of the lumen of pregnant women with early-onset and late-onset severe preeclampsia.Results:The tube wall thickness [ (119.69±8.31) μm], systolic blood pressure [ (162.86±4.94) mmHg], diastolic blood pressure [ (103.09±2.35) mmHg], and 24-hour urine protein [ (2.17±0.31) g/24 h] in the early preeclampsia group were higher than those in the late preeclampsia group [ (101.04±5.78) μm, (146.95±6.43) mmHg, (92.13±4.74) mmHg, (1.62±0.23) g/24 h] and the normal pregnancy group [ (99.82±5.56) μm, (116.42±9.31) mmHg, (74.25±6.74) mmHg, (0.06±0.02) g/24 h], the placental tissue lncRNA TGFB2-AS1 level (0.62±0.16), lumen area [ (133.74±20.16) μm 2], gestational week of delivery [ (32.15±1.74) weeks], weight of the newborns [ (2.25±0.26) g] were lower than those in the late-onset preeclampsia group [ (0.99±0.21), (185.49±22.75) μm 2, (36.14±1.59) weeks, (3.37±0.32) g] and the normal pregnancy group [ (1.02±0.23), (186.42±23.71) μm 2, (38.19±1.56) weeks, (3.42±0.37) g] ( P<0.05). The systolic blood pressure, diastolic blood pressure, and 24-hour urine protein in the late preeclampsia group were higher than those in the normal pregnancy group, gestational week of delivery was lower than the normal pregnancy group ( P<0.05). Placental tissue lncRNA TGFB2-AS1 of pregnant women with early-onset severe preeclampsia was positively correlated with the lumen area, and negatively correlated with the thickness of the tube wall ( P<0.05). There was no correlation between lncRNA TGFB2-AS1 and the lumen area and wall thickness in the placental tissue of pregnant women with late-onset severe preeclampsia ( P>0.05) . Conclusion:The lncRNA TGFB2-AS1 expression in the placenta tissue of pregnant women with early-onset severe preeclampsia is abnormally low, which may be related to the insufficient recasting of the placental spiral artery.

Objective To compare the effects of smoking and non smoking on postoperative pain of laparoscopic cholecystectomy. Methods Sixty patients having underwent selective laparoscopic cholecystectom were divided into smoking group and non smoking group by random digits table with 30 cases each. In smoking group, 14 cases quitted smoking within 1 week before operation. The Fagerstrom test of nicotine dependence (FTND) was evaluated before operation in smoking group, and FTND ≥ 6 scores was in 11 cases. The visual analog score (VAS), Bruggrmarm comfort score (BCS), sedation-agitation score (SAS), immediately, 15 min, and 30 min after entering postanesthesia care unit (PACU) and leaving PACU was evaluated. The operation time, anesthesia time, wake up time, extubation time, PACU time, using rate of remedial measures and untoward reaction were recorded. Results There were no statistical differences in operation time, anesthesia time, wake up time, extubation time, SAS and incidence of untoward reaction between 2 groups (P>0.05). The PACU time and using rate of remedial measures in smoking group were significantly higher than those in non smoking group:(39.7 ± 5.1) min vs. (31.3 ± 6.1) min and 30.0% (9/30) vs. 0, and there were statistical differences (P<0.05). The VAS immediately, 15 min and 30 min after entering PACU and leaving PACU in smoking group was significantly higher than that in non smoking group: (2.90 ± 0.85) scores vs. (1.00 ± 0.83) scores, (2.70 ± 0.47) scores vs. (0.73 ± 0.69) scores, (2.60 ± 0.56) scores vs. (1.13 ± 0.73) scores, (2.23 ± 0.57) scores vs. (1.13 ± 0.73) scores; and the BCS was significantly lower than that in non smoking group:(1.80 ± 0.61) scores vs. (2.90 ± 0.99) scores, (1.90 ± 0.31) scores vs. (2.87 ± 1.00) scores, (2.10 ± 0.31) scores vs. (2.47 ± 0.82) scores, (2.17 ± 0.38) scores vs. (2.47 ± 0.82) scores, and there were statistical differences (P<0.05). The VAS immediately after entering PACU in patients of FTND ≥ 6 scores was significantly higher than that in patients of FTND<6 scores:(3.6 ± 0.7) scores vs. (2.5 ± 0.7) scores, the BCS was significantly lower than that in patients of FTND <6 scores:(1.5 ± 0.5) scores vs. (2.0 ± 0.6) scores, and there were statistical differences (P<0.05). The VAS immediately after entering PACU in patients of non- quit smoking was significantly higher than that in patients of quit smoking: (3.4 ± 0.7) scores vs. (2.4 ± 0.6) scores, and there were statistical differences (P<0.05). Conclusions Smokers have more severe postoperative pain in laparoscopic cholecystectomy and higher postoperative opioid requirement than nonsmokers. Quit smoking before surgery will reduce postoperative pain and related complications. Appropriate increase of analgesic drugs can prevent postoperative pain in patients with smoking.

Objective To investigate the effect of CaMK Ⅱ expression on apoptosis of rat hepatocytes BRL-3A.Methods Rat BRL-3A cells were stable passage were cultured.The CaMK Ⅱ γ protein (LV-CaMK Ⅱ γ group) and CaMK Ⅱ γshRNA (shRNA group) lentiviral expression systems were constructed.The corresponding blank vectors (LV-NC group and shRNA-NC group) and normal saline (CON group) were perfused into the control groups.The expression levels of CaMK Ⅱ,Cyt C and MF proteins were detected by Western blotting,and the apoptosis rate of BRL-3A cells was measured by Tunel method.Results The protein expression of CaMK Ⅱ,Cyt C and AIF in LV-CaMK Ⅱ γ group was significantly higher than that in CON group (P＜0.05).The protein expression of CaMK Ⅱ,Cyt C and AIF in shRNA group was significantly lower than that in CON group (P＜ 0.05).There was no significant difference among CON group,LV-NC group and shRNA-NC group (P＞0.05).At the same time point,the apoptosis rate of hepatocytes in LV-CaMK Ⅱ γ group was significantly higher than that in CON group (P＜0.05).At the same time point,the apoptosis rate of hepatocytes in shRNA group was significantly higher than in CON group (P＜0.05).There was no significant difference in the apoptosis of hepatocytes among CON group,LV-NC group and shRNA-NC group (P＞0.05).Conclusion The specific CaMK Ⅱ signaling pathway can inhibit the apoptosis of BRL-3A cells,while the enhanced CaMK Ⅱ signaling pathway promotes the apoptosis of BRL-3A cells.

PURPOSE: The aim of our study was to explore the relationships between the M2 isoform of pyruvate kinase (PKM2) and the sensitivity of human non-small cell lung cancer (NSCLC) cells to docetaxel in vitro. MATERIALS AND METHODS: With the method of plasmid transfection, we silenced the expression of PKM2 successfully in A549 and H460 cells. Western blotting and real-time PCR were applied to detect PKM2 expression at protein and gene levels. Cell viability was examined by CCK8 assay. Cell cycle distribution and apoptosis were examined by flow cytometry. P21 and Bax were detected. RESULTS: Expression of PKM2 mRNA and protein were significantly decreased by shRNA targeting PKM2. Silencing of PKM2 increased docetaxel sensitivity of human NSCLC A549 and H460 cells in a collaborative manner, resulting in strong suppression of cell viability. The results of flow cytometric assays suggested that knockdown of PKM2 or docetaxel treatment, whether used singly or in combination, blocked the cells in the G2/M phase, which is in consistent with the effect of the two on the expression of p21. Cells with PKM2 silencing were more likely to be induced into apoptosis by docetaxel although knockdown of PKM2 alone can't induce apoptosis significantly, which is in consistent with the effect of the two on Bax expression. CONCLUSION: The results suggest that PKM2 knockdown could serve as a chemosensitizer to docetaxel in non-small lung cancer cells through targeting PKM2, leading to inhibition of cell viability, increase of cell arrest of G2/M phase and apoptosis.
Apoptosis ; Blotting, Western ; Carcinoma, Non-Small-Cell Lung ; Cell Cycle ; Cell Line* ; Cell Survival ; Drug Therapy ; Flow Cytometry ; Humans* ; In Vitro Techniques* ; Lung Neoplasms ; Methods ; Plasmids ; Pyruvate Kinase* ; Pyruvic Acid* ; Real-Time Polymerase Chain Reaction ; RNA, Messenger ; RNA, Small Interfering* ; Transfection

Objective To evaluate the effect of comprehensive intervention on perioperative antimicrobial prophy-laxis in clean incision surgery in a hospital.Methods From 2011 ,clean incision surgery cases were performed com-prehensive intervention,antimicrobial use in 2011 -2013 were compared.Results A total of 5 945 cases of clean in-cision surgeries were investigated between 2011 and 2013,3 827 cases (64.37%)received prophylactic use of anti-microbial agents.Prophylactic antimicrobial usage rates in 2011 -2013 were 84.95%,69.99%,and 52.97% re-spectively(χ2 =380.94,P <0.001);the correct rates of medication time were 50.97%,79.99%,and 98.95% re-spectively(χ2 =827.02,P <0.001 );the percentages of prophylactic antimicrobial use ≤24 hours were 24.91 %, 39.96%,and 64.95% respectively(χ2 =422.55,P <0.001 );additional antimicrobial usage rates during surgery were 50.00%,60.00%,and 80.00% respectively(χ2 =59.47,P <0.001 ).Conclusion The implementation of comprehensive intervention measures can standardize antimicrobial use,reduce prophylactic antimicrobial usage rate,improve the correct rate of medication time,shorten the duration of antimicrobial use,and implement addition-al use of antimicrobial agents during surgery.

Objective To explore the role of pyruvate kinase M2 (PKM2) siRNA in the radiosensitivity of human lung cancer A549 cells.Methods PKM2 siRNA was synthesized according to the coding sequence of PKM2 mRNA and then was transferred into A549 cells with lipofectamine.The expressions of PKM2 gene and protein was detected by RT-PCR and Western blot,respectively.The experiments were divided into PKM2 siRNA interference group,siRNA negative control group,and blank control group.The cells of each group were exposure to 6 MV X-rays in different dose.Radiosensitivity was evaluated by colony formation assay.Flow cytometry was applied to analyze cell cycle distribution and apoptosis.Data are representative of three independent experiments.Results Ccompared with blank control cells,the expressions of PKM2 gene and protein in the PKM2 siRNA transferred A549 cell was efficiently diminished (t =20.91,47.00,P ＜0.01) with inhibition rates of (70.27 ± 1.38)％ and (70.42 ± 1.18) ％,respectively.Compared with control,PKM2 siRNA transfection significantly decreased the D0,Dq,N and SF2 values (t =43.82,28.44,15.60,29.63,P ＜ 0.01) and hence yield a sensitization enhancement ratio (SER) of 1.27.In addition,the percentage of G2/M phase cells in the siRNA group and irradiated group were both significantly higher than that of the blank control group (t =8.35,27.87,P ＜ 0.01).The combined treatments of PKM2 siRNA interference and irradiation arrested more cells in the G2/M phase compared to either treatment alone.The apoptosis rate of siRNA group was not dramatically different from that of blank control group.The apoptosis rate of irradiation group was higher than that of blank control group (t =23.99,P ＜ 0.01),and the combined treatments of siRNA and irradiation enhanced the apoptotic rate compared to either treatment alone (t=9.42,65.21,P ＜ 0.01).Conclusions Specific blockage of PKM2 expression by gene silencing could enhance the sensitivity of human lung cancer A549 cells to radiotherapy in vitro,which may due to the cell cycle arrest and apoptosis induction after irradiation.

BACKGROUND:At present, the proteome is a mature technology that has been applied in basic research fields related to liver transplantation. But, it has been not reported in research related to reduced-size liver transplantation.
OBJECTIVE:To explore the expression of differential proteins related to hepatic energy metabolism fol owing reduce-size liver transplantation in rats by using by proteomic technology.
METHODS:The improved model of reduced-size liver transplantation was used in this experiment. The donor was health female Lewis rats and the recipient was male Wistar rats for liver transplantation. The difference between the donor and the recipient was about 20 g. The weight of donor liver/the weight of recipient donor was approximately equal to 50%. The donor liver tissue was harvested and trimmed to the required size. The portal vein and infrahepatic vena cava were cannulated, and the biliary tract was implanted into the donor bile duct for transplantation. Then the donor was transplanted into the recipient after the removal of original liver tissue. Hepatic specimens were harvested by 1, 3 and 7 days after reduced-size liver transplantation. Then, the harvested specimens were compared with the normal donor and recipient liver tissue that were previously harvested and frozen, to generate two-dimensional gel electrophoresis profile using proteome technology. Then tandem mass spectrometry and databases analysis were performed after two-dimensional electrophoresis for identifying differential protein stains.
RESULTS AND CONCLUSION:In this experiment, 72 differential protein stains with over lo-fold changes were selected. After identification, 32 proteins showed clear functions, and among them three differential proteins (ATP synthase beta subunit, electron-transferring flavoprotein beta peptide and proton-transferring ATP synthase) were involved in the process of cel energy metabolism. The proteins were distributed on 1 and 7 days after reduce-size liver transplantation, accounting for 6%.

Objective To discuss the differential expression of hepatic stress proteins after reduced-size liver transplantation in rats.Methods The specimens of liver tissues were procured on 1 d,3 d and 7 d after the improved model of reduced-size liver transplantation in rats.Then,the two-dimensional electrophoresis of these specimens was compared with that of the original liver tissues of normal donors and recipients.The differentially expressed protein spots were selected with the standard of change times greater than 10 or less than 1 /10 and then were analyzed and identified by mass-spectrometric technique and data bases.Results Seventy-two differentially expressed protein spots were found in total.And the 32 kinds of proteins were identified with definite function through mass spectrometry and a series of identifications.The expression difference of heat shock protein-8 and hypertrophy agonist reactive protein was larger,amounting 7% (5 /72)of all differential proteins.Conclusions This study provides fundamental research data for studying the relation between liver ischemia-reperfusion injury after liver transplant and the above differential proteins of stress reaction in transplant liver which are found after reduced-size liver transplantation in rats.

Objective To explore the effect of nano-hydroxyapatite/collagen scaffolds incorporating ADM-PLGA microspheres in repairing large bone defects of rabbit femoral condyle. Methods Animal models of bone defects were induced in 21 New Zealand white rabbits by drilling holes in bilateral femoral lateral condyles , and the rabbits were equally divided into 3 groups:group A as the control group with the defects untreated , group B treated by filling with nano-hydroxyapatite/collagen scaffolds (NHAC), and group C treated by filling with the nano-hydroxyapatite/collagen scaffolds incorporating ADM-PLGA microspheres (ADM-PLGA-NHAC). At week 12 after implanting , the rabbits were all sacrificed for the implanted scaffolds , which were then examined by X-ray , and Micro-CT 3D reconstruction and in histology for evaluation of the new bone formation. Results X-ray, Micro-CT and the measurement and analysis of BMD indicated thatthere was no significant differencein the new bone formation between group B and group C (P > 0.05). The histological examination revealed that. 12 weeks after operation an evident number of new born bones were seen on the implanted scaffolds in groups B and C , while very few were seen scattering in group A. Conclusion The nano-hydroxyapatite/collagen scaffolds incorporating ADM-PLGA microspheres is effective in repairing bone defect without influencing the prosthetic process.