Objective To study the relationship between in situ telomerase activity and the expressin of bcl-2 and p53 proteins in human ovarian epithelial tumors(OETs), and explore their effect in pathogenesis of ovarian cystadenocarcinoma. Methods The telomerase activity of ovarian epithelial tumors was measured by in situ telomerase activity labeling (ISLT), and bcl-2 and p53 expression was detected by SP immunohistochemical method. Results ⑴The positive rates of telomerase in ovarian cystadenocarcinomas(OCAC), their surrounding ovarian tissues(SOT),borderline cystadenomas(BCA),and cystadenomas(CA)were 92 3%(24/26), 0(0/26), 42 8%(9/21) and 0(0/15) respectively, and the positive rate was significantly higher in OCAC than that in SOT, BCA and CA(P0 05); ⑵The positive rates of Bcl-2 in OCAC, SOT, BCA and CA were 65 38%(17/26), 0(0/26), 52 38%(11/21) and 26 66%(4/15) respectively, which were significantly higher in OCAC and BCA than those in SOT and CA(P0 05). Conclusion The results indicated that overexpression of bcl-2 protein may be related to telomerase activation, the telomerase activation induced by bcl-2 overexpression may result in malignant transformation of ovarian epithelials, and p53 mutation may not affect telomerase activity during ovarian cystadenocarcinogenesis.

Objective To investigate expressions of vascular endothelial growth factor (VEGF) and nm23 proteins and microvessel density in HCC,simultaneously,their relation with metastasis was also studied.Methods The expression of VEGF and nm23 and CD34 protein in 56 HCCs were detected by immunohistochemical technique.Results Of 56 HCCs, the intensity of VEGF expression in HCCs with metastasis was significantly higher than that in HCCs without metastasis (P

Objective To investigate the hepatocyte targeted specific property of galactosylated chitosan-graft-polyethyleneimine (GC-PEI)/DNA complexes in vitro and in vivo.Methods With the plasmid expressing enhanced green fluorescent protein (pEGFP-C1) as the reporter gene,the formation of GC-PEI/DNA complexes was induced to self-assemble in 0.01 mol/L phosphate buffered saline(PBS),150 mmol/L NaCl,or 5% glucose solution (GS).The complexes were characterized by the particle size,Zeta potential,DNA binding and protection capacity,and further tested for cytotoxicity and hepatocyte targeted degradation of DNaseⅠand the serum,which presented as a well-formed sphere or compacted nucleocapsid structure at a diameter of 50-200 nm.The GC-PEI copolymer showed no obvious toxicity in the tested cell lines.Acute toxicity assay revealed that the mice grew well in 2 weeks with GC-PEI dosage from 50 to 300 μg.The assay by flow cytometry and fluorescent microscope showed that the transfection efficiency in hepatocyte lines (L02,QSG7701/core) was higher than that in non-hepatocyte lines (SGC7901,HBE) in vitro.In vivo,the GFP was obviously expressed in the liver tissue and not expressed in other organs 48 h after the transfection.Conclusion GC-PEI copolymer may carry the exogenous gene specifically to hepatocytes in vitro and in vivo,which has very good liver targeted specific property.

Objective: To explore the guiding significance of pulse contour cardiac output (PiCCO) monitoring technology in the treatment of fluid replacement during shock stage of extensive burn in clinic. Methods: Sixty-five patients with extensive burn hospitalized in our unit from January 2014 to December 2018, conforming to the inclusion criteria, were recruited to conduct a prospective controlled research. According to the order of admission, 35 odd-numbered patients and 30 even-numbered patients were enrolled in routine rehydration group (25 males and 10 females) and PiCCO monitoring rehydration group (21 males and 9 females) respectively, with the age of (48±9) and (44±8) years respectively. All patients of the two groups were rehydrated according to the rehydration formula of the Third Military Medical University during shock stage. The rehydration speed was adjusted in routine rehydration group according to the general indexes of shock such as central venous pressure, mean arterial pressure, heart rate, respiratory rate, urine volume, and clinical symptoms of patients. PiCCO monitoring was performed in patients of PiCCO monitoring rehydration group, and the global end-diastolic volume index combined with the other relevant indicators of PiCCO were used to guide rehydration on the basis of the monitoring indicators of routine rehydration group. The heart rates and positive fluid balance volumes at post injury hour (PIH) 8, 16, 24, 32, 40, 48, 56, 64, and 72, the diuretic dosage at PIH 48 and 72, the total fluid replacement volumes, urine volumes, blood lactic acid, platelet count, and hematocrit at PIH 24, 48, and 72, the length of intensive care unit (ICU) stay, and the incidence of complications and death within 28 days after injury were compared between patients in the two groups. Data were processed with analysis of variance for repeated measurement, t test, Bonferroni correction, Mann-Whitney U test, chi-square test, and Fisher′s exact probability test. Results: The heart rates of patients in the two groups were similar at PIH 8, 16, 24, 32, 40, 48, and 56 (t=0.775, 1.388, 2.511, 2.203, 1.654, 2.303, 1.808, P>0.05), and the heart rates of patients in PiCCO monitoring rehydration group at PIH 64 and 72 were obviously lower than those of routine rehydration group (t=3.229, 3.357, P<0.05 or P<0.01). The positive fluid balance volumes of patients in the two groups were similar at PIH 8, 16, 40, and 56 (t=0.768, 1.670, 2.134, 2.791, P>0.05), and the positive fluid balance volumes of patients in PiCCO monitoring rehydration group at PIH 24, 32, 48, 64, and 72 were obviously less than those of routine rehydration group (t=3.364, 4.047, 2.930, 2.950, 2.976, P<0.05 or P<0.01). The amount of diuretics used by patients in the two groups was similar at PIH 48 and 72 (Z=-0.697, -1.239, P>0.05). The total fluid replacement volumes of patients in PiCCO monitoring rehydration group at PIH 24, 48, and 72 were (13 864±4 241), (9 532±2 272), and (8 480±2 180) mL, respectively, obviously more than those in routine rehydration group [(10 388±2 445), (8 095±1 720), and (7 059±1 297) mL, respectively, t=-3.970, -2.848, -3.137, P<0.05 or P<0.01]. The urine volumes of patients in the two groups at PIH 24 were close (t=-1.027, P>0.05). The urine volumes of patients in PiCCO monitoring rehydration group at PIH 48 and 72 were (3 051±702) and (3 202±624) mL respectively, obviously more than those in routine rehydration group [(2 401±588) and (2 582±624) mL respectively, t=-4.062, -4.001, P<0.01]. The levels of blood lactate acid of patients in PiCCO monitoring rehydration group at PIH 24, 48, and 72 were obviously lower than those in routine rehydration group (t=4.758, 6.101, 3.938, P<0.01). At PIH 24 and 48, the values of the platelet count of patients in PiCCO monitoring rehydration group were obviously higher than those in routine rehydration group (t=-2.853, -2.499, P<0.05), and the values of hematocrit of patients in PiCCO monitoring rehydration group were obviously lower than those in routine rehydration group (t=2.698, 4.167, P<0.05 or P<0.01). Both the platelet count and hematocrit of patients in the two groups were similar at PIH 72 (t=-1.363, 0.476, P>0.05). The length of ICU stay of patients in PiCCO monitoring rehydration group was obviously shorter than that of routine rehydration group (t=2.184, P<0.05). Within 28 days after injury, the incidence of complications of patients in routine rehydration group was obviously higher than that in PiCCO monitoring rehydration group (P<0.05), while the mortality rate of patients in routine rehydration group was similar to that in PiCCO monitoring rehydration group (P>0.05). Conclusions The application of PiCCO monitoring technology in monitoring fluid replacement in patients with extensive burn can quickly correct shock, reduce the occurrence of organ complications caused by improper fluid replacement, and shorten the length of ICU stay, which is of great significance in guiding the treatment of burn shock.

OBJECTIVE: To identify aberrantly expressed miRNAs in rectal cancer. METHODS: We used the miRCURY™ Array® LNA microRNA chip (v.14.0) to evaluate miRNA expression levels between rectal cancer tissues and adjacent non-tumor tissues; an average change more than 2-fold (and P value less than 0.05) was set as a cutoff level. All 6 paired rectal cancers were classified pathology stage C or D. RESULTS: Eighty-eight miRNAs were up-regulated and 46 miRNAs have been reported in colorectal cancer; 40 miRNAs were down-regulated in rectal cancers and 15 miRNAs have been reported in colorectal cancer. To compare the relative miRNA expression levels as measured by RT-qPCR and chip analysis, we analyzed expression levels of these miRNAs in the cancer tissues. The results showed that miRNA expression (increased or decreased) in the paired benign and tumor tissue was consistent between the two methods in all cases. Expression levels of 6 up-regulated miRNAs (by chip analysis compared to RT-qPCR) varied in a range from -11.9% to 39.1% . Expression levels of 5 down-regulated miRNAs varied in a range from 1.4% to 29.4%. The Pearson correlation of relative miRNAs expression levels was analyzed by cDNA array versus RT-qPCR, and found to be 0.96 (P<0.01). CONCLUSION miRNA profile in rectal cancer showed unique characteristics, and identified a series of new, aberrantly expressed miRNAs.
Aged ; Colorectal Neoplasms ; genetics ; metabolism ; Female ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Humans ; Male ; MicroRNAs ; genetics ; metabolism ; Middle Aged ; Real-Time Polymerase Chain Reaction ; Up-Regulation

OBJECTIVETo explore the neuro-protective effect and mechanism of qingkailing injection (QKL) against cerebral injury caused by E. coli-meningitis (CM).

METHODSThe CM model rabbits were treated by ampicillin with QKL as adjuvant. The leukocyte count and protein content in cerebral spinal fluid (CSF), the contents of water, sodium, potassium and calcium in cerebral tissues were measured before, 16 h and 26 h after Bacillus coli injection respectively. The expression of matrix metalloproteinase-9 (MMP-9) was determined at the same time.

RESULTSAdjunctive treatment with QKL can not only inhibit the increase of leukocyte cells, protein content in CSF, and water, sodium, calcium content in cerebral tissues, but also the decrease of potassium content revealed during simple antibiotic treatment. It also can decrease the expression of MMP-9 in cerebral tissues of rabbits with CM.

CONCLUSIONAs an adjunctive treatment, QKL can prevent transient inflammatory reaction and aggravation of brain injury in CM induced by simple antibiotic treatment, its mechanisms might relate with calcium antagonism and attenuation of MMP-9 expression in brain tissues.

Ampicillin ; therapeutic use ; Animals ; Anti-Bacterial Agents ; therapeutic use ; Brain ; metabolism ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Injections ; Male ; Matrix Metalloproteinase 9 ; biosynthesis ; Meningitis, Escherichia coli ; drug therapy ; Neuroprotective Agents ; therapeutic use ; Phytotherapy ; Rabbits

OBJECTIVETo study the effect of hepatitis C virus nonstructural protein NS(3) (HCV NS3) on telomerase activity and carcinogenesis.

METHODSStreptavidin-peroxidase (SP) conjugated method was used to detect the expression of HCV NS(3) protein in NIH3T3 cells transfected with plasmid pRcHCNS(3)-5' and pRcHCNS(3)-3'. Telomerase activity was detected by an in situ telomerase activity labeling method, telomeric repeat amplification protocol polymerase chain reaction (TRAP-PCR) and telomerase PCR enzyme linked immunosorbent assay (ELISA) technology in the transfected and non-transfected NIH3T3 cells.

RESULTSHCV NS(3) protein was expressed in the NIH3T3 cells transfected with plasmid pRcHCNS(3)-5' expressing HCV NS(3) C-terminal deleted protein or with plasmid pRcHCNS(3)-3' expressing HCV NS(3) N-terminal deleted protein. The positive signal of HCV NS(3) protein was localized in the cytoplasm of NIH3T3 cells, and the signal intensity of the former was stronger. Telomerase activity in NIH3T3 cells transfected with plasmid pRcHCNS(3)-5' was stronger than that in NIH3T3 cells transfected with plasmid pRcHCNS(3)-3' (P < 0.01), whereas telomerase activity in NIH3T3 cells transfected with plasmid pRcCMV or untreated NIH3T3 cells was weaker than that in NIH3T3 cells transfected with plasmid pRcHCNS(3)-3' (P < 0.05). The expression level of HCV NS(3) protein was significantly correlated with the strength of telomerase activity (P < 0.05). The results obtained by in situ telomerase activity labeling corresponded to the results by telomerase PCR ELISA technology.

CONCLUSIONSHCV NS(3) protein may activate telomerase through endogenous mechanism to induce host cell transformation. The effect of HCV NS(3) C-terminal deleted protein on telomerase activity in the host cell may be stronger than that of HCV NS(3) N-terminal deleted protein. In situ telomerase activity labeling was a reliable technology for studying pathological morphology and telomerase activity in tissues and cells.

3T3 Cells ; Animals ; Enzyme-Linked Immunosorbent Assay ; methods ; Mice ; Plasmids ; genetics ; Polymerase Chain Reaction ; methods ; Telomerase ; genetics ; metabolism ; Transfection ; Viral Nonstructural Proteins ; genetics ; physiology

OBJECTIVE: To explore the effect of Fuzheng Huayu (FZHY) recipe on the fenestration of capillarization in liver sinusoidal endothelial cells (LSECs).
 METHODS: Ten Sprague Dawley (SD) rats were fed with 0.46 g/kg FZHY powder by intragastric administration. Two hours later, a second gavage were given to the rats. The serum from rat heart at 1 hour after second gavage was collected (FZHY group, n=10). Another ten SD rats was administrated with distilled water through the same process and served as the control (control group, n=10). The serum from both groups were separately diluted with Dulbecco minimum essential medium (DMEM) for 10% and served as the culture medium for LSECs. At the different conditions, the vWF and CD31 expressions were detected by immunocytochemistry and Western blot, while the changes of LSECs fenestrae structure were observed under scanning electron microscopy.
 RESULTS: 1) Immunocytochemistry and Western blot showed that the vWF and CD31 protein levels were lower in LSECs in the FZHY group than those in the control group. The gray levels of vWF and CD31 protein were 0.548±0.020 and 0.262±0.010 in the FZHY group, and 0.845±0.090 and 0.383±0.010 in the control group respectively, with statistical significant difference (t=5.18, 9.61, both P<0.05). 2) The results from scanning electron microscopy showed that the fenestration of LSECs was closed and almost lost in the control group, but many fenestra appeared in the LSECs in the FZHY group.
 CONCLUSION FZHY recipe can suppress the expression of vWF and CD31, increase the fenestrae on the LSECs surface and reverse the capillarization of LSECs.
Animals ; Drugs, Chinese Herbal ; pharmacology ; Endothelial Cells ; cytology ; drug effects ; Liver ; cytology ; Platelet Endothelial Cell Adhesion Molecule-1 ; chemistry ; Rats ; Rats, Sprague-Dawley ; von Willebrand Factor ; chemistry

OBJECTIVE: To explore the disease spectrum for abnormal 3-hydroxyisovalerylcarnitine (C5OH) metabolism identified through newborn screening and clinical diagnosis patients and the key points for differential diagnosis so as to raise the awareness of pediatricians for such diseases. METHODS: Clinical data of 85 neonates with abnormal C5OH metabolism identified from February 2004 to January 2022 at Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine were collected. Their clinical manifestations and results of tandem mass spectrometry (MS/MS), gas chromatography mass spectrometry (GC-MS) and genetic testing were retrospectively analyzed. RESULTS: Among the 85 cases, 46 (54.1%) were identified by neonate screening, whilst 39 (45.9%) were clinically diagnosed patients. Five diseases were diagnosed, including 28 cases with multiple carboxylase deficiency (MCD, 32.9%), 29 cases with 3-methylcrotonyl-coenzymeAcarboxylasedeficiency (MCCD, 34.1%), 4 cases with 3-methylglutaconic acid (3-MGA, 4.7%), 7 cases with 3-hydroxy-3-methylglutaric acid (3-HMG, 8.2%), and 17 cases with beta-ketothiolase deficiency (BKD, 20.0%). The disorders were characterized by sudden onset, anorexia, vomiting, diarrhea, abnormal breathing, consciousness disorder, spasm and developmental delay. CONCLUSION Among newborns with abnormal C5OH metabolism, MCCD is the most common disorder, which was followed by BKD and MCD. For patients with abnormal C5OH metabolism, MCD is the most common, followed by BKD and 3-HMG. C5OH related diseases have great heterogeneity. Combination of blood acylcarnitine levels, urinary organic acid levels and genetic testing based on clinical characteristics can help to attain the diagnosis.
Humans ; Infant, Newborn ; China ; Neonatal Screening ; Retrospective Studies ; Tandem Mass Spectrometry/methods*