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 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

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.

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 clinical effects of fiberoptic bronchoscopy airway lavage (FBAL) in the treatment of extremely severe burn patients with severe inhalation injury.Methods:From January 2015 to January 2019, 47 extremely severe burn patients with severe inhalation injury who were hospitalized in Tongren Hospital of Wuhan University & Wuhan Third Hospital, meeting the inclusion criteria, were recruited in this retrospective cohort study. According to whether or not they were treated with FBAL, the patients were divided into fiberoptic bronchoscopy group (23 cases, 19 males and 4 females) and routine group (24 cases, 20 males and 4 females), with the age of (44±11) and (49±9) years, and the admission time of 4 (3, 4) h and 4 (3, 5) h respectively. The patients in routine group were given routine comprehensive treatment, and the patients in fiberoptic bronchoscopy group were treated with FBAL on the basis of routine comprehensive treatment. The pH value, arterial partial pressure of oxygen (PaO 2), arterial partial pressure of carbon dioxide (PaCO 2), arterial oxygen saturation (SaO 2), oxygenation index, white blood cell count (WBC), neutrophils, blood lactic acid, and procalcitonin (PCT) at admission and on post injury day (PID) 3, 5, 7, and 10, the time of mechanical ventilation, the day of intensive care unit (ICU) stay, the incidence of complications and death within PID 28 were compared between the two study groups. The occurrences of bronchospasm and asphyxia of patients in fiberoptic bronchoscopy group were monitored. Data were statistically analyzed with independent sample t test, Mann-Whitney U test, Bonferroni correction, chi-square test, and Fisher′s exact probability test. Results:(1) At admission, the values of pH, PaO 2, PaCO 2, SaO 2, and oxygenation index of patients in the two groups were similar ( Z=-0.118, -0.320, -0.362, -2.416, -0.234, P>0.05). On PID 3, 5, 7, and 10, the values of pH, PaO 2, SaO 2, and oxygenation index of patients in fiberoptic bronchoscopy group were significantly higher than those of routine group ( Z3 d=-4.711, -4.161, -5.525, -2.661; Z5 d=-3.489, -4.678, -5.875, -3.599; Z7 d=-5.104, -4.619, -5.876, -4.844; Z10 d=-4.026, -5.698, -5.877, -4.716; P<0.05 or P<0.01). The PaCO 2 values of patients in fiberoptic bronchoscopy group were significantly lower than those of routine group ( Z=-2.895, -3.162, -3.407, -2.831, P<0.05 or P<0.01). (2) At admission and on PID 3, 5, and 7, the values of WBC, blood lactic acid, and PCT of patients in the two groups were similar ( Z=-0.830, -0.915, -0.458, -0.648, -1.714, -1.479; -0.330, -0.128, -1.766, -0.494, -1.396, -1.522, P>0.05). On PID 10, the values of WBC, blood lactic acid, and PCT of patients in fiberoptic bronchoscopy group were significantly lower than those of routine group ( Z=-3.502, -2.630, -2.662, P<0.05 or P<0.01). At admission, the value of neutrophils of patients in fiberoptic bronchoscopy group was 0.887 (0.862, 0.912), which was similar to 0.887 (0.856, 0.897) in routine group ( Z=-0.404, P>0.05). On PID 3, 5, 7, and 10, the values of neutrophils of patients in fiberoptic bronchoscopy group were respectively 0.848 (0.802, 0.867), 0.831 (0.815, 0.849), 0.798 (0.771, 0.849), 0.796 (0.751, 0.869), which were significantly lower than those of routine group [0.882 (0.820, 0.906), 0.871 (0.835, 0.903), 0.845 (0.819, 0.905), 0.881 (0.819, 0.916), Z=-2.756, -2.810, -2.618, -3.033, P<0.05]. (3) The time of mechanical ventilation and the days of ICU stay of patients were shorter in fiberoptic bronchoscopy group than those in routine group ( Z=-2.199, t=2.368, P<0.05). Within PID 28, the number of patients with complications was significantly less in fiberoptic bronchoscopy group than in routine group ( χ2=5.436, P<0.05), while the incidence of death within PID 28 in fiberoptic bronchoscopy group was similar to that of routine group ( P>0.05). The airway lavage procedures of patients in fiberoptic bronchoscopy group went well with no bronchospasm or asphyxia occurred. Conclusions:FBAL is effective in treating extremely severe burn patients combined with severe inhalation injury. It can improve the oxygenation status of the lung, reduce the systemic inflammatory reaction of patients, shorten the time of mechanical ventilation and ICU stay, and reduce the incidence of complications.

Objective:To investigate the clinical effects of expanded flaps in reconstructing scar contracture deformities in the face and neck after extensive burns.Methods:A retrospective observational study was conducted. From May 2016 to September 2022, 17 patients with scar contracture deformities in the face and neck after extensive burns were admitted to Tongren Hospital of Wuhan University & Wuhan Third Hospital, including 13 males and 4 females, aged 23 to 55 years, with 3 patients having degree Ⅱ cervical contracture, 14 patients having degree Ⅲ cervical contracture, and 12 patients having facial scar contracture deformity. In the first stage, 34 rectangular skin and soft tissue expanders (hereinafter referred to as expanders) with rated capacity of 100-600 mL were inserted into the face, chest, shoulder, and abdomen, and then the normal saline was injected for expansion. In the second stage, the scar tissue was removed and the contracture was released to correct the deformity. Two expanded facial flaps were transplanted in local fashion, 17 expanded flaps were transplanted in pedicled fashion, and 15 expanded flaps were freely transplanted to repair the secondary wounds after release, with artery pressurization was performed in 7 flaps. Indocyanine green fluorescence imaging was used to evaluate the arterial blood perfusion and venous return of the flaps during transplantation. The incision area of 32 flaps except 2 facial flaps was 10 cm×8 cm-36 cm×16 cm. The wounds of 31 flap donor sites were closed by direct suture, and the wound of 1 flap donor site was repaired by autologous split-thickness scalp transplantation. The skin condition of inserted place, expansion time, and total amount of normal saline injection of expanders, complications of skin and soft tissue expansion surgery, and survival of flap after the second stage surgery were observed and recorded. The long-term face and neck reconstruction effect and recovery of flap donor area were followed up. At the last follow-up, the 5-level Likert scale was used to evaluate the efficacy satisfaction of patients.Results:Of the 34 expander inserted places in 17 patients, 22 places were superficial scar skin after deep partial-thickness burns, 8 places were superficial scar skin after multiple skin donations, and 4 places were normal skin. After 4 to 15 months of expansion, the total normal saline injection volume was 238 to 2 000 mL, with no complications occurred. After the second stage surgery, the distal part of 2 pedicled flaps was partially necrotic, and the necrotic wounds were healed after flap dressing and free transplantation of contralateral expanded triangular flaps, respectively; the other flaps survived completely. During 6 to 18 months of follow-up, except for 2 expanded paraumbilical flaps and 1 expanded groin flap, which were bloated and improved by flap thinning, the appearance and texture of the other flaps were good, and all the flap donor sites recovered well. At the last follow-up, the face and neck scar contracture deformities were significantly improved in all patients, and the satisfaction of curative effect of patient was very satisfactory in 8 patients and relatively satisfactory in 9 patients.Conclusions:The expanded flaps of chest, abdomen, and other parts, combined with local advance, pedicled, and free transplantation, can effectively reconstruct scar contracture deformities in the face and neck after extensive burns, restore the function of operative area and improve the appearance simultaneously, with high degree of patient satisfaction, which is worthy of promotion in clinic.

Objective:To explore the risk factors for atrial fibrillation in adult patients with critically severe burns after the first surgery.Methods:This study was a retrospective case series study. From January 1, 2018 to March 31, 2023, 211 adult patients with critically severe burns were admitted to the Department of Burns of Tongren Hospital of Wuhan University & Wuhan Third Hospital and met the inclusion criteria, including 158 males and 53 females, aged 24-81 years. According to whether atrial fibrillation occurred after the first surgery, the patients were divided into postoperative atrial fibrillation (POAF) group (23 cases) and non-POAF group (188 cases). The following indexes of patients in POAF group were collected, including the onset time, duration, treatment method, and number of patients with more than once of atrial fibrillation after the first surgery. The following data of the two groups of patients were collected, including general data, such as gender, age, burn type, total burn area, full-thickness burn area, inhalation injury, underlying diseases, mechanical ventilation, and sepsis; electrolyte imbalance and blood index level before the first surgery; the first surgery-related information such as surgical length and surgical method; volume changes and vital signs during the first surgery, such as total volume of fluid infusion, total volume of blood transfusion, volume of blood loss, hypotension, and hypothermia; postoperative hypothermia; inflammatory index levels before the first surgery and on the first day after the first surgery, such as procalcitonin levels, white blood cell count, neutrophil count, lymphocyte count, platelet count, neutrophil to lymphocyte ratio (NLR), platelet count to lymphocyte ratio (PLR); mortality within 30 days of admission. The independent risk factors for occurrence of atrial fibrillation in adult patients with critically severe burns after the first surgery were screened.Results:The onset time of atrial fibrillation of patients in POAF group was 2 (2, 4) hours after the first surgery, and the duration of atrial fibrillation was 16 (6, 26) hours. Twenty-one patients were treated with intravenous injection of amiodarone, two patients were treated with cardiac electrical cardioversion, and atrial fibrillation of all patients converted to sinus rhythm after treatment. Three patients experienced atrial fibrillation more than once. The age was 59 (42, 70) years and the total burn area was 90% (70%, 94%) total body surface area (TBSA) in patients in POAF group, which were significantly higher than 48 (38, 56) years and 70% (60%, 83%) TBSA in non-POAF group (with Z values of -2.64 and -3.56, respectively, P<0.05). Compared with those in non-POAF group, the incidence rate of inhalation injury of patients in POAF group was significantly higher ( χ2=4.45, P<0.05), the total volumes of fluid infusion and blood transfusion during the first surgery were significantly increased (with Z values of -3.98 and -3.75, respectively, P<0.05), the incidence rates of hypothermia during the first surgery and hypothermia after the first surgery were significantly increased (with χ2 values of 8.24 and 18.72, respectively, P<0.05), the levels of procalcitonin before the first surgery and on the first day after the first surgery, as well as the NLR on the first day after the first surgery were significantly increased (with Z values of -3.03, -2.19, and -2.18, respectively, P<0.05), the lymphocyte count (with Z values of -2.07 and -2.60, respectively, P<0.05) and platelet count (with Z values of -3.35 and -3.58, respectively, P<0.05) were significantly reduced before the first surgery and on the first day after the first surgery, and the mortality rate within 30 days of admission was significantly higher ( χ2=4.03, P<0.05). There were no statistically significant differences in other indexes between the two groups of patients ( P>0.05). Multivariate logistic regression analysis showed that age, total burn area, and intraoperative hypothermia were independent risk factors for the occurrence of atrial fibrillation in adult patients with critically severe burns after the first surgery (with odds ratios of 1.08, 1.07, and 4.18, 95% confidence intervals of 1.03-1.12, 1.03-1.11, and 1.48-11.80, respectively, P<0.05). Conclusions:Age, total burn area, and intraoperative hypothermia are independent risk factors for the occurrence of atrial fibrillation in adult patients with critically severe burns after the first surgery. Patients with atrial fibrillation have an increased risk of death.