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Objective:To investigate the influence of preoperative thyroid dysfunction on the 30-day postoperative mortality and complications in elderly patients with hip fracture.Methods:A retrospective analysis was conducted of the 349 elderly patients with hip fracture who had been admitted to Department of Orthopedic Trauma, Beijing Luhe Hospital Affiliated to Capital Medical University from January 2018 to December 2019. They were 108 males and 241 females, with an average age of 76.3 years (from 60 to 104 years). There were 190 femoral intertrochanteric fractures and 159 femoral neck fractures. By the preoperative level of thyroid function, the patients were divided into a normal function group of 290 cases and a dysfunction group of 59 cases. The 2 groups were compared in terms of hospital stay, mortality and incidence of complications within 30 days postoperation.Results:In this cohort, the rate of 30-day postoperative mortality was 3.4%(12/349) and the incidence of 30-day postoperative complications 14.6%(51/349). The 2 groups were comparable because there was no significant difference between them in the preoperative general data except for the preoperative comorbidity of coronary heart disease ( P>0.05). In the dysfunction group, the hospital stay averaged (10.2±6.9) d, the rate of 30-d postoperative mortality 1.7%(1/59) and the incidence of 30-day postoperative complications 16.9%(10/59), which were insignificantly different from those in the normal function group [(10.7±7.5) d, 3.8%(11/290) and 14.1%(41/290), respectively] ( P> 0.05). Conclusion:Since preoperative thyroid dysfunction does not affect the 30-day postoperative mortality and postoperative complications in the elderly patients with hip fracture but no definite thyroid disease, routine thyroid function screening is not recommended for them.
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Objective:To explore the expression of miR-125 in nasopharyngeal carcinoma tissues and the possible regulatory mechanism of biological characteristics of tumor cells.Methods:Thirty cases of carcinoma and paracancerous tissues from patients with nasopharyngeal carcinoma admitted to Southern Hospital of Sixth People′s Hospital Affiliated to Shanghai Jiaotong University from June 2018 to June 2019 were collected. The expressions of miR-125 and fibroblast growth factor 2 (FGF-2) mRNA were detected by fluorescent quantitative PCR. Nasopharyngeal carcinoma CNE-2 cells were transfected by miR-125 mimic (miR-125 mimic group), and negative control group was set (NC group). Transwell chamber assay was used to determine cell invasion ability, scratch healing assay was used to determine cell migration ability, WST-1 assay was used to assess cell viability, flow cytometry and electron microscopy were used respectively to detect apoptosis and autophagy, dual luciferase reporter assay was used to analyze the target of miR-125, and Western blotting was used to detect the expressions of related proteins.Results:The relative expression of miR-125 mRNA in nasopharyngeal carcinoma tissues was 0.692±0.316, which was significantly lower than 1.501±0.748 in the adjacent tissues ( t=5.242, P<0.001). The relative expression of FGF-2 mRNA in nasopharyngeal carcinoma tissues was 1.317±0.552, which was significantly higher than 0.783±0.241 in the adjacent tissues ( t=7.360, P<0.001). The miR-125 mRNA expression of CNE-2 cells in the miR-125 mimic group was 4.091±0.145, which was significantly higher than 0.993±0.137 in the NC group ( t=85.062, P<0.001). The proliferative activities of CNE-2 cells in the miR-125 mimic group at 48 and 96 h after transfection were significantly lower than those in the NC group (0.891±0.214 vs. 1.295±0.245, t=6.802, P<0.001; 0.934±0.208 vs. 1.488±0.269, t=8.924, P<0.001). The number of transmembrane cells and cell migration rate of the miR-125 mimic group were 36 000±3 820 and (39.4±6.5)%, which were significantly lower than 74 000±7 500 and (102.7±10.6)% of the NC group ( t=24.728, P<0.001; t=27.883, P<0.001). The apoptosis rate of CNE-2 cells in the miR-125 mimic group was (22.5±1.4)%, which was significantly higher than that in the NC group (1.4±0.5)% ( t=77.740, P<0.001). The relative expression of the apoptotic protein Bax in the miR-125 mimic group was 0.983±0.158, which was significantly higher than that in the NC group (0.418±0.122; t=15.503, P<0.001), and the relative expression of Bcl-2 was 0.688±0.174, which was significantly lower than that of the NC group (1.013±0.109; t=8.670, P<0.001). Autophagy was observed in miR-125 overexpressing CNE-2 cells by electron microscopy, and the relative expression ratio of autophagy protein LC3Ⅱ/LC3Ⅰ in the miR-125 mimic group was 2.517±0.209, which was significantly higher than 1.238±0.135 in the NC group ( t=28.156, P<0.001). The expression of FGF-2 protein in the miR-125 mimic group was 0.504±0.118, which was significantly lower than 1.228±0.134 in the NC group ( t=22.210, P<0.001). The double luciferase report confirmed FGF-2 as the target gene of miR-125. At 12, 24, 48 and 96 h after the transfection, the cell proliferative activities of CNE-2 cells co-transfected by FGF-2 gene plasmid and miR-125 mimic were significantly higher than those of CNE-2 cells transfected by miR-125 mimic (all P<0.05), and the apoptosis rate was significantly lower than that of CNE-2 cells transfected by miR-125 mimic [(6.2±1.5)% vs. (17.6±2.4)%, t=22.062, P<0.001]. Conclusion:The expression of miR-125 is down-regulated in nasopharyngeal carcinoma tissues. Overexpression of miR-125 may inhibit the proliferation, migration and invasion of nasopharyngeal carcinoma CNE-2 cells and promote the apoptosis and autophagy of CNE-2 cells by down-regulating FGF-2 expression.
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OBJECTIVE： To investigate the vasodilatory effect mechanism of psoralen and bakuchiol. METHODS： The rat thoracic aorta was isolated to prepare vascular rings and de-endothelium vascular rings. Using contraction rate as index， the intact endothelium or de-endothelium vascular rings were pre-incubated with N-nitro-L-arginine methyl ester （L-NAME， 100 μmol/L）； vasodilatory effect of low-dose， medium-dose and high-dose of psoralen or bakuchiol（0.1，1，10 μmol/L）on aortic vessels pre- contracted with norepinephrine （NE， 1 μmol/L） or potassium chloride （KCl， 60 mmol/L） were investigated. The de-endothelium vascular rings were pre-incubated with calcium dependent potassium channel inhibitors tetraethylammonium chloride （TEA， 0.1 mmol/L） and inward rectifying potassium channel inhibitor barium chloride （BaCl2，0.1 mmol/L）； vasodilatory effect of low-dose， medium-dose and high-dose of bakuchiol （0.1， 1， 10 μmol/L） on de-endothelium vascular vessels pre-contracted with NE （1 μmol/L） were investigated. The microvascular endothelial cells were isolated by collagenase-neutral protease digestion； the effects of low-dose， medium-dose and high-dose of psoralen or bakuchiol （0.1， 1， 10 μmol/L） on the expression of eNOS protein were studied by ELISA. RESULTS： Psoralen and bakuchiol could significantly reduce the contraction rate of endothelium-intact aortic rings pre-contracted with NE（P＜0.01）； medium-dose and high-dose of psoralen and bakuchiol could significantly reduce the contraction rate of endothelium-intact aortic rings pre-contracted with KCl（P＜0.05 or P＜0.01）； while the contraction rate could be increased by de-endothelium and NOS inhibition significantly （P＜0.05 or P＜0.01）. The medium-dose and high-dose of bakuchiol could significantly reduce the contraction rate of de-endothelium vascular vessels pre-contracted with NE （P＜0.05 or P＜0.01）. The contraction rate could be increased by inhibiting inward rectifier potassium channels in vascular smooth muscle （P＜0.01）. Different dosages of psoralen and bakuchiol could significantly increase the expression levels of eNOS protein in rat cardiac microvascular endothelial cells（P＜0.01）. CONCLUSIONS： Psoralen and bakuchiol may play a role in vasodilation via endothelium-dependent NO pathway and by promoting eNOS protein expression in endothelial cells； bakuchiol may play a role in vasodilation via non-endothelium dependent pathway as opening inward rectifying potassium channel.
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Objective UPLC-MS/MS bio-analysis method was developed for the simultaneous determination ofberberine,naringin,hesperidin,and neohesperidin in plasma of rats.Methods UPLC Acquity BEH C18 (50 rmm × 2.1 mm,1.7 μm) column was used,mobile phases were containing 0.05％ formic acid and 2 mmol/L ammonium formate in water (A)-containing 0.05％ formic acid in acetonitrile (B) as the mobile phase gradient elution;SD rats were randomly divided into oral administration berberine group,Citrus aurantium extract group,and berberine and C.aurantium extract compatibility group.Results UPLC-MS/MS method could be applied to determination of berberine,naringin,hesperidin,and neohesperidin,method validation meets the requirements of biological sample analysis.When rats were administered with berberine and C.aurantium extract compatibility,the plasma concentration of berberine was much more than single dose of berberine group and the bioavailability of berberine was increased.Meanwhile,naringin and neohesperidin can be detected in rat's plasma.Conclusion The bioavailability of flavonoids is significantly improved as well compared to the single dose of C.aurantium extract.This suggests that berberine and C.aurantium extract compatibility has significant drug-drug interaction.
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Objective To establish a RP-HPLC method investigate the processing technique and mechanism of Eucommiae Cortex.Methods The RP-HPLC method was applied to simultaneously determining six ingredients,geniposidic acid,geniposide,genipin,chlorogenic acid,(+)-pinoresinol-di-β-D-glucopyranoside,and(+)-syringaresinol-di-β-D-glucopyranoside,in the different processed barks of Eucommia ulmoides.Results The valid method with good accuracy could be well used to study the processing technique of E.ulmoides;Besides,target ingredients in E.ulmoide were decreased within 6 h when they were processed.Conclusion Established RP-HPLC is a reliable method which could be used to research the processing technique of the barks ofE.ulmoides.Moreover,the result of this study could be provided with significant evidence of processed barks of E.ulmoides.
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BACKGROUND: Human keratinocyte growth factor-2 (hKGF-2) has extensive physiological functions, which plays an important role in embryonic development, tissue-repairing, nervous regeneration, vascularization and development of tumor.OBJECTIVE: To clone hKGF-2 gene, obtain the expression of hKGF-2 in Escherichia coli(E.coli) and determine its bioactivity, so as to provide experimental basis for further investigation.DESIGN: Open experiment.SETTING: Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention.MATERIALS: The experiment was conducted in State Key Laboratory of Viral Genetic Engineering, Institute for Viral Disease Control and Prevention of Chinese Center for Disease Control and Prevention. The temperature control expression vector pBV220 was constructed by State Key Laboratory of Viral Genetic Engineering; EcoR Ⅰ , BamH Ⅰ , T4 DNA ligase (Promega Co., Ltd.); The specific polymerase chain reaction (PCR) of hKGF-2 (Manufactured by Shanghai Boya Biotechnology Co., Ltd.); Heparin-Sepharose CL-6B (Pharmacia Company); PCR rapid purification kit,Trizol kits for total RNA extract, Kits for RT-PCR (GIBCO Co., Ltd.); Kits for rapid extraction of plasmid DNA (Boda Company); BL-21-codon plus compent cells (Stratagene Co., Ltd.).METHODS: High-expression strain BL-21 codon plus competent cells was used to express and purify initially recombinant hKGF-2 protein, and its activity was detected. RT-PCR was adopted to obtain hKGF-2 cDNA from lung tissues of naturally aborted fetus and clone it into pBV220 carri er plasmid. The hKGF-2 protein expressed in BL-21 codon plus competent cells of E.coli. Affinity chromatography and ion exchange chromatography were applied in isolation and purification, and the bioactivity of expression protein was determined in cell proliferation test.MAIN OUTCOME MEASURES: The length and sequence of cDNA segment in hKGF-2, the expression of hKGF-2 gene inE.coli and the purification of hKGF-2 activity.RESULTS: The segment of hKGF-2 cNDA was about 500 bp, and hKGF-2 protein highly expressed in BL-21, which had soluble expression in the supernatant. SDS-PAGE showed that the relative molecular mass was about 20000, and hKGF-2 protein could significantly promote the mitotic activity of NIH3T3 cells. The A value (490 nm) of hKGF-2 in the 1 μg/L, 5 μg/L and 10 μg/L groupswere higher than that in the blank control group, and the differences were significant (which were 0.174±0.022,0.220±0.029,0.306±0.050,0.066±0.004 respectively,P ＜ 0.001).CONCLUSION: hKGF-2 gene is successfully cloned, which highly expresses in BL-21 of the E.coli. Purified hKGF-2 protein can stimulate the proliferation of NIH3T3 cells and significantly promote its mitotic activity.
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This study studied the use of ERCP and nasobiliary tube in the diagnosis of fungal infection of biliary tract and the efficacy of combined use of local administration via nasobiliary tube and intravenous antifungal treatment for severe biliary tract fungal infection. 5 patients in our series,with age ranging from 47 to 68 y (mean 55.8), were diagnosed as having mixed bacterial and fungal infection of biliary tract as confirmed by smear or/and culture of bile obtained by ERCP and nasobiliary drainage. Besides routine anti-bacteria therapy, all patients received local application of fluconazole through nasobiliary tube and intravenous administration of fluconazole or itraconazole in terms of the results of in vitro sensitivity test. The mean duration of intravenous fluconazole or itraconazole was 30 days (24-40 days), and that of local application of fluconazole through nasobiliary drainage tube was 19 days (8-24 days). During a follow-up period of 3-42 months, all patient's fungal infection of biliary tract was cured. It is concluded that on the basis of typical clinical features of biliary tract infection, fungal detection of smear/culture of bile obtained by ERCP was the key for the diagnosis of fungal infection of biliary tract. Local application antifungal drug combined with intravenous anti-fungal drugs might be an effective and safe treatment for fungal infection of biliary tract.
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Objective To observe prospectively the role of endoscopic diagnosis and treatment of biliary leakages in patients with liver transplantation, and the incidence of bile duct stricture after healing of the leakage. Methods Six eases of T-tube leakage and seven cases of anastomosis leakage complicating liver transplantation were enrolled in this prospective study. Six patients treated by endoscopic plastic stent placement , 2 by naso-biliary catheter drainage, 2 by papillosphincterotomy and 3 by naso-biliary catheter drainage combined with plastic stent placement. Some patients received growth hormone treatment. Results The bile leak resolution time was between 10-35 days in 10 patients with complete document. The median time of leak resolution was 15. 3 days. Four cases of anastomosis stricture, three cases of common hepatic duct and one ease of multiple bile duct stenosis were observed by followed-up nasobiliary catheter cholangiography or ER-CP. Conclusion Endoscopic nasobiliary catheter or plastic stent placement is a safe and effective treatment for bile duct stricture occurred after bile leak resolution in most of liver transplantation patients. Naso-biliary catheter combined with plastic stent placement maybe the best choice for treating bile leak, because, theoretically, it may prevent serious condition happened at accidental nasobiliary catheter dislocation, and it may have prophylactic effect on upcoming bile duct stricture and should be further confirmed.
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Objective Survivin is a recently described inhibitor of apoptosis. It is undetectable in normal tissue of adults, but abundantly expressed in transformed cells and a variety of human cancers. This study was aimed to investigate the effect of antisense oligonucleotide (ASODN) targeting survivin on hepatic cell apoptosis, proliferation and sensitivity to chemotherapeutic drugs. Methods ASODN targeting survivin was designed and constructed. Cultured cells were divided into 6 groups: control group, lipofectin group, oligonucleotide(ODN) group, 200 nmol/L ASODN group, 400 nmol/L ASODN group and 600 nmol/L ASODN group. After transfection for 20 h, cultured cells were harvested to carry on the next tests. Cell morphological change was observed with invert microscope; survivin protein expression was detected by Western blot; apoptotic index (AI) and proliferative index (PI) were examined by flow cytometry; rate of inhibition (IR) by chemotherapeutic drugs was determined by the colorimetric MTT cell viability/proliferation assay. Results Expression of survivin in ASODN groups was significantly decreased compared with that in the control, lipofectin and ODN group; abnormal morphological change of cells was observed in ASODN transfected groups; the AI of ASODN groups was significantly higher than that of other groups; the PI of ASODN groups was significantly lower than that of other groups; the IR of chemotherapeutic drugs in ASODN groups was significantly higher than that of other groups. Conclusions Expression of survivin may decrease in hepG2 cells after ASODN transfection. ASODN targeting survivin can induce cell apoptosis, inhibit cell proliferation and sensitize hepatocarcinoma cells to chemotherapy.
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Objective Survivin, one of the inhibitory protein of apoptosis, is selectively expressed in the tissue of malignant tumors. The present study was to investigate the role of Survivin expression in the pathogenesis of hepatocellular carcinoma (HCC). Methods Two HCC cell lines, forty histologically verified HCC specimens and their neighboring noncancerous tissues were obtained. Western blotting was used to test protein expression of Survivin. Semi quantitative RT PCR was used to measure Survivin mRNA levels. TUNEL was employed to examine the apoptosis index of HCC tissues. Results Survivin protein and mRNA expression were detected in 2 HCC cell lines and 34 of 40 (85.0%) HCC tissues. In contrast, no expression of Survivin in corresponding noncancerous tissues was detected. The positivity rate of Survivin expression in cases of intrahepatic metastasis was 93.5%, higher than that without intrahepatic metastasis (55.6%, P
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Objective To investigate telomere length and cellular DNA content in different gastric lesion mucosa, and their relation with gastric mucosal carcinogenesis. Methods Telomere length were determined by southern hybridization. Cellular DNA content was detected by flow cytometry. Results Telomere length in intestinal metaplasia (IM) grade 2 was significantly shorter than that in normal gastric, IM grade 0 or grade 1. Telemere length of gastric carcinoma cells was the shortest in all of the biopsy specimens. Telomere length ratio in patients of corresponding surrounding nontumorous tissues with IM grade 2 was the largest in 45 resected gastric carcinoma. In flow cytometry, The aneuploid of gastric carcinoma (n=18), chronic atrophic gastritis (CAG) contained IM grade 2 (n=8), grade 1 (n=40), grade 0 (n=20), chronic superficial gastritis (CSG n=46) and normal gastric mucosa (n=10) was 33.3%,12.5%,10.0%,0.0%,0.0% and 0.0%, rspectively, in all of the biopsy specimens. In 45 resected gastric carcinoma specimens, Telomere length of 18 aneuploid was significantly shorter than that of 27 diploid. Furthermore reverse correlation was observed between telomere length and the DNA index in 18 aneuploid. Conclusions Telomere length were shortened as normal mucosa changed into intestinal metaplasia and more into gastric cancer. The normal and CSG mucosa shows no aneuploid. The positivity of DNA aneuploid tends to increase with the progression of intestinal metaplasia. In addition, telomere length and the DNA index show a reverse correlation. It is speculated that the shorter the telomere length the more amplificative activity the DNA. Telomere length and increased DNA index may be a predictor of stomach carcinogenesis.
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Objective To analyze and discuss the utility of parenteral nutrition in inflammatory bowel disease (IBD).Methods In the light of the diagnostic criteria of IBD by the National Conference in 1993 as to the chronic noninfectious bowel disease,the inpatients of IBD in our department from Jan.1986 to Dec.1996 were analyzed,and the medical treatment of IBD and the role of parenteral nutrition evaluated.Results It was demonstrated that the annual average number of hospitalized patients had increased in the past 11 years,and the parenteral nutrition resulted in a better outcome,in addition to SASP or 5-ASA.Conclusion It was believed that following the principal treatment of IBD, nutritional support is also essential.