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Biomedical subjects

Jing Zhao

Publications and source records attributed to Jing Zhao.

At least 109 records · Page 6Linked to original sources

[Pro-apoptotic efficiencies of three reconstructed human caspase-8 on cervical cancer cell line HeLa].

BACKGROUND & OBJECTIVE: Apoptosis is a kind of evolutional high conservative cell death. Transferring high active pro-apoptotic molecules into cancer cells to induce apoptosis is a potential strategy for cancer gene therapy. Based on our previous generation of reconstructed human caspase-8, which can continuously induce apoptosis of cervical cancer cell line HeLa, by reversing its large and small subunits, this study was designed to investigate the pro-apoptotic efficiencies of 3 reconstructed human caspase-8 (Casp8CD, Rev8, and Rev8L) on HeLa cells, and to explore the feasibility of reconstructed human caspase-8 as potential apoptosis-inducing candidates. METHODS: The eukaryotic expression vectors pIRES2-EGFP carrying Casp8CD, Rev8, and Rev8L genes were transfected into HeLa cells, and breast cancer MCF-7 cells. Expressions and pro-apoptotic effects of Casp8CD, Rev8, and Rev8L genes were observed under fluorescent microscope, and their pro-apoptotic efficiencies were assessed by MTT assay and cells counting. The flexibilities of linking-peptides between subunits of Rev8 and Rev8L were analyzed by bioinformatics. RESULTS: Expressions of the 3 reconstructed caspase-8 genes were observed under fluorescent microscope, and the HeLa and MCF-7 cells expressing Rev8 or Rev8L genes displayed typical apoptotic volume decrease (AVD). MTT assay showed that compared with control cells, A(570) values of Rev8- and Rev8L-transfected cells began to decrease 20 h after transfection. Cell counting results indicated that cell death ratio of Casp8CD-, Rev8-, and Rev8L-transfected cells were 16.9%, 52.3%, and 47.7%, respectively, 24 h after transfection; and 12.9%, 51.6%,and 61.2%,respectively,48 h after transfection. Bioinformatics analysis showed that the linking-peptides between subunits of Rev8 and Rev8L were flexible. CONCLUSION: Rev8 and Rev8L molecules have similar pro-apoptotic effects and efficiencies, but over-expressed Casp8CD had no significant pro-apoptotic effects.

Apoptosis↗

Protein secondary structure prediction using dynamic programming.

In the present paper, we describe how a directed graph was constructed and then searched for the optimum path using a dynamic programming approach, based on the secondary structure propensity of the protein short sequence derived from a training data set. The protein secondary structure was thus predicted in this way. The average three-state accuracy of the algorithm used was 76.70%.

Algorithms↗

[Preliminary study on nasal spray of interferon alpha-2b used for prevention of rubella and measles virus infections].

OBJECTIVE: To evaluate the efficacy of the interferon alpha-2b nasal spray in prevention of rubella and measles virus infections. METHODS: The properly selected volunteer groups have been divided into interferon alpha-2b experimental and control group. The experimental group received interferon alpha-2b treatment by nasal spray for 2 days before the immunization, then both groups were challenged with rubella and measles attenuated live vaccine respectively through nasal spray. The sera from pre-immunization and 21 and 28 days after immunization were collected to test the IgG antibody titers. The influence on the viral antibody titer reflects the viral preventive effect by interferon alpha-2b. RESULTS: The antibody titer difference of measles virus between experimental and control group was 1.26 (21 day) and 2.96 (28 day), there were statistically difference between them; the difference of rubella virus was 0.95 (21 day) and 0.37 (28 day), but there were no statistically differences found. CONCLUSION: The preliminary results showed that the interferon alpha-2b can be used as prevention method for measles and rubella viral infections.

Administration, Intranasal↗

[Determination of positive bronchodilating response using impulse oscillation system in children].

OBJECTIVE: Impulse oscillation system (IOS) as an approach to lung function determination is suitable for children, especially for preschool children in that it only requires the individual to be examined to breathing stably with tidal volume. However, until now there have been no uniformly agreed criteria for positive bronchial reversibility test in clinical practice. A screening method was applied to seek for answer when this question in the hope of providing objective evidence for clinical diagnosis of respiratory diseases, esp. asthma. METHODS: One hundred and fifty-six children patients during asthma attack were recruited randomly for this study from January to December, 2004. These patients included 103 boys and 53 girls aged from 5 to 18 years with mean age of 8.84 +/- 2.58 years, who visited the doctors in the Clinical and Educational Centre for Asthma, Capital Institute of Pediatrics and were diagnosed as asthma. Maximal expiratory flow volume and IOS lung function were determined followed by bronchodilator reversibility test in all patients. The sensitivity and specificity of IOS parameters, total respiratory impedance (Zrs), respiratory resistance at 5 Hz (R5) and reactance at 5 Hz (X5) for diagnosing asthma at different improvement levels after inhalation of bronchodilator were calculated respectively using forced expiratory volume in first second (FEV(1)) and maximal mid-expiratory flow (MMEF) as gold standard. And the regression equation between parameters of lung function which were determined with the two different methods was analyzed. RESULTS: After inhalation of bronchodilator, the lung function of the children was improved significantly (P < 0.01), with the FEV(1), MMEF increase and Zrs, R5, and X5 decrease. There were significantly positive correlations (P < 0.01) between the improvement rate of parameters of lung functions determined with the two methods, and the highest correlation coefficients existed between X5 and FEV(1) and MMEF (respectively 0.676, 0.571), correlation coefficients between X5 and Zrs ranked second (0.519 and 0.505, respectively). When an increase of FEV(1) equal to or greater than 15%, or increase of MMEF equal to or greater than 30% was used as reference for positive bronchial reversibility, the sensitivity and specificity were relatively high for considering positive bronchial reversibility test with at least 20% decrease of Zrs, R5, and 30% decrease of X5 with IOS lung function. The sensitivity and specificity of Zrs, R5 and X5 calculated from FEV(1) were 0.62, 0.51; 0.49, 0.54; 0.70, and 0.54, respectively. The sensitivity and specificity of Zrs, R5 and X5 calculated from MMEF were 0.63, 0.54; 0.53, 0.60; 0.60, and 0.58, respectively. The regression equation showed that the decrease of Zrs, R5 and X5 corresponding to 15% increase of FEV(1) and 30% increase of MMEF were 21.7%, 21.3%; 19.9%, 19.5%; 30.1%, and 29.6%, respectively. CONCLUSION: When lung function is determined with IOS in children, only when decrease of Zrs and R5 is equal to or more than 20% and decrease of X5 is 30% or more after inhalation of bronchodilator, can the bronchial reversibility test be considered as positive.

Adolescent↗

[Spectral properties, protonation and fluorescence quantum yield of ciprofloxacin].

Fluorescence spectra, ultraviolet absorption spectra, and protonation of Ciprofloxacin (CIP) at different pH values have been studied. Fluorescence quantum yield of CIP under neutral condition has been measured. In HCl medium with [H+] > 1 mol x L(-1), CIP molecules (simplified as HL) may accept three protons to exist as H4L3+ with very weak fluorescence, and its maximum fluorescence emission wavelength (lambdamax) is 456 nm. In acidic solution of pH 0 to pH 2, CIP mainly exists as H3L2+ form with lambdamax at 450 nm, and fluorescence intensity is relatively weak and increases with increasing of pH. In the range of pH 2 to pH 4, CIP mainly exists as H2L+ form with a strong fluorescence, and lambdamax is still at 450 nm. When pH>4, lambdamax gradually blue-shifts to 414 nm, fluorescence intensity slightly decreases as pH increases, and at the same time an evident change in ultraviolet absorption spectrum is observed, indicating that H2L+ has lost proton to exist as dipole ion form HL. When pH>8, the fluorescence intensity decreases until disappearance as pH increases, indicating that HL has lost proton to exist as non-fluorescence anion ion form L-. In the molecular form changing process, the maximum excitation wavelength of CIP is essentially constant at 275 nm, but the maximum emission wavelength changes obviously. In a buffer solution with pH 7.0, and using quinine bisulphate as a reference, the fluorescence quantum yield of CIP at maximum excitation wavelength 275 nm was measured to be 0.12.

Algorithms↗

[Multilayer amniotic membrane transplantation for treatment of necrotizing herpes simplex stromal keratitis].

OBJECTIVE: To evaluate the efficacy of multilayer amniotic membrane transplantation (AMT) combined with antivirus and corticosteroid drug to treat necrotizing herpes simplex stromal keratitis. METHODS: Thirteen patients (13 eyes) of necrotizing stromal keratitis were referred to Shandong Eye Institute and Qingdao Eye Hospital between January 2003 and April 2004. The course of disease was 3 - 22 months (mean 15 months). Corneal inflammation persisted and corneal ulcer progressed despite topical and systemic antiviral treatment for over 1 weeks. Multilayer amniotic membrane transplantation was performed after excluding of bacterial and fungal infection by microbiologic studies including smears and cultures of necrotic corneal tissue and confocal microscope. Topical and systemic antiviral medications were given with adjuvant corticosteroid eyedrops postoperatively. We investigated the healing of corneal ulcer and improvement of stromal edema with slit lamp biomicroscope, the integrity of corneal defect with fluorescein staining, the migration of healthy corneal epithelial cells and transformation of amniotic membrane with confocal microscopy. All patients were followed up for 3 - 13 months (mean 10 months). RESULTS: Corneal ulcer healed within 1 - 3 weeks postoperatively with negative fluorescein staining. Corneal stromal edema faded away within 1 month. Superficial amniotic membrane patches dissolved or shed on postoperative day 7 - 10, while the deeper grafts were adhered into the ulcer and fused with the surrounding fibroblasts. One of these grafts remained in situ more than 3 months. Confocal microscope examination indicated flat epithelial progenitor cells on the surface of residual amniotic membrane. Corneal transparence was achieved in 7 eyes, macula in 4 eyes and leucoma in 2 eyes 3 months after the operation. No recurrence of necrotizing stromal keratitis was occurred in 13 patients during the follow-up period. CONCLUSION: Multilayer AMT combined with antivirus and corticosteroid treatment is an effective method to treat necrotizing herpes simplex stromal keratitis.

Acyclovir↗

Forced-air warming and fluid warming minimize core hypothermia during abdominal surgery.

OBJECTIVE: To evaluate the effect of intraoperative combined forced-air warming and fluid warming system on patient's core temperature, blood loss, transfusion demand, extubation time, and incidence of postoperative shivering. METHODS: Forty patients with American Society of Anesthesiologists physical status I and II, aged 18-70 years, scheduled for elective abdominal surgery were randomly assigned to receive intraoperative warming from a forced-air blanket and fluid warming system or conventional cotton blanket, 20 in each group. The core temperature was recorded every 20 minutes during the operation, as well as the blood loss, blood transfusion, extubation time, and incidence of postoperative shivering. RESULTS: The core temperature at the end of the surgery in the warming group was significantly different from that in the control group (36.4 +/- 0.4 degrees C vs. 35.3 +/- 0.5 degrees C, P < 0.001). Application of intraoperative warming significantly shortened the time between the end of the surgery and extubation (P < 0.01). Postoperative shivering occurred in 30% of the patients in the control group compared to no patient in the warming group (P < 0.01). CONCLUSION: Active warming with air-forced blanket and fluid warming system provides sufficient heat to prevent hypothermia during abdominal surgery.

Abdomen↗

Salusins promote cardiomyocyte growth but does not affect cardiac function in rats.

Salusin-alpha and -beta are newly found polypeptides that stimulate proliferation, hypotension and bradycardia in vascular smooth muscle cells (VSMCs) and fibroblasts. Propresalusin mRNA is widespread, and positive stains for salusins have been observed in many human tissues such as endothelium and ventricular tissue. To investigate the bio-effect of salusins on cardiovascular function, 20 nmol/kg salusin-alpha or 2 nmol/kg salusin-beta was intravenously (i.v.) injected into rats, and isolated rat hearts were perfused with 10(-12) to 10(-7) mol/l salusin-alpha or -beta. (45)Ca(2+) uptake and (3)H-Leucine incorporation were determined in cultured neonatal rat cardiomyocytes. Neither salusin-a nor -beta affected cardiac function in vivo or in vitro but salusin-beta decreased mean arterial blood pressure (MAP). The polypeptides' stimulation of (45)Ca(2+) uptake and (3)H-Leucine incorporation was concentration-dependent, and the incorporation was inhibited by nicardipine (Nic) and FK-506 [FK; an inhibitor of calcineurin (CaN)]. PD(98059) [PD; inhibitor of mitogen-activated protein kinase (MAPK)] and chelerythrine [inhibitor of protein kinase C (PKC)] inhibited salusin-stimulated (3)H-Leucine incorporation. Endothelin-1 (ET) synergistically increased salusin-induced (45)Ca(2+) uptake. Our results suggest that salusin-alpha and -beta did not directly affect cardiac function in the rat heart but that they improved calcium uptake and protein synthesis in neonatal rat cardiomyocytes through the calcium, calcineurin, MAPK and PKC signal pathways. Salusins may be regulatory factors for myocardial growth and hypertrophy.

Adenosine Triphosphatases↗

Effects of adrenomedullin on aldosterone-induced cell proliferation in rat cardiac fibroblasts.

Aldosterone induces cardiac remodeling in cardiovascular diseases by stimulating the proliferation, production and secretion of collagen in fibroblasts. It also stimulates vascular smooth muscle cells to produce and secrete adrenomedullin (ADM), which has a cytoprotective effect against cardiovascular damage. We examined the effect of aldosterone on ADM production and secretion in rat cardiac fibroblasts, and the effect of ADM on aldosterone-stimulated fibroblast proliferation to observe the interaction between endogenous ADM and aldosterone. We detected ADM produced and secreted from cultured cardiac fibroblasts and the intracellular cAMP level by radioimmunoassay; evaluated cell proliferation by the level of [3H]-thymine incorporation; measured preproADM gene expression by reverse transcriptase polymerase chain reaction (RT-PCR); and monitored extracellular signal related kinase (ERK) activity by the phosphorylation of myelin basic protein in the presence of [gamma-32P] ATP. Our results showed that aldosterone-stimulated secretion of ADM and its mRNA expression were concentration-dependent, which could be inhibited by the specific antagonist of mineralocorticoid receptor, spironolactone. In contrast, ADM inhibited aldosterone-induced fibroblast proliferation and ERK activity. Treatment with ADM24-50 (a new antagonist of specific ADM receptors) and calcitonin gene-related peptide (CGRP)8-37 (the antagonist of CGRP receptor type 1), to attenuate the action of endogenous ADM, reinforced the aldosterone-induced proliferation and inhibited the intracellular cAMP production stimulated by aldosterone. Thiorphan, an inhibitor of ADM degradation, inhibited the [3H]-thymine incorporation and reinforced the intracellular cAMP level induced by aldosterone. We reach the conclusion that aldosterone stimulates rat cardiac fibroblasts to produce and secrete ADM, which in turn regulates the proliferation-induced effects of aldosterone in these cells.

Adrenomedullin↗

High pressure study of Ru3(CO)12 by X-ray diffraction, Raman, and infrared spectroscopy.

The single-crystal X-ray structure of Ru(3)(CO)(12) is reported at 8 pressures ranging from 1 atm (0.0 GPa) to 8.14(5) GPa. Although intramolecular bonding parameters remain relatively constant, intramolecular and intermolecular nonbonding contact distances decrease by an average of 4% and 15%, respectively. At 8.14 GPa, O...O, C...O, and C...C intermolecular distances as short as 2.54(4), 2.64(6), and 3.07(4) A, respectively, are observed, and the unit cell compresses to 75% of the ambient pressure volume. Raman and infrared spectroscopic measurements show that carbonyl stretching frequencies shift to higher wavenumber values by as much as 80 cm(-)(1), even though Ru-C and C-O distances stay roughly constant throughout the entire pressure range studied. Compression of the sample to above 18 GPa with laser radiation results in an irreversible transformation due to either decomposition or a total collapse of D(3)(h) molecular geometry accompanied by color darkening.

Journal Article↗

[The effect of intraoperative warming on patient core temperature].

OBJECTIVE: To investigate the influence of using fluid warming and forced-air warming system on patient core temperature, blood loss, blood transfusion, extubation time, and postoperative shivering. METHODS: Forty ASA (American Society of Anesthesiologists' Physical Status) I-II patients, aged 21-69 years, scheduled for elective abdominal surgery under general anesthesia, were enrolled in the study. The patients were premedicated with intramuscular dolantin 50 mg and atropine 0.5 mg. Anesthesia was induced with midazolam 1 mg, fentanyl 50-100 microg and propofol 1.5-2.0 mg/kg. Tracheal intubation was facilitated with vecuronium 1mg and succinylcholine 1.5-2.0 mg/kg. The patients were mechanically ventilated and anesthesia was maintained with isoflurane 1.5-2.0%, 50% N2O in oxygen and intermittent iv boluses of fentanyl (total dose 5-6 microg/kg). Vecuronium was used for muscle relaxation during maintenance of anesthesia. The patients were randomly divided into 2 groups: control group (n = 20) and warming group (n = 20). In both groups, the patients were covered with surgery blanket. In the warming group, patients were additionally warmed with fluid warming device and forced-air warming system during the operation. The core temperature was recorded every 20 minutes during the operation, as well as the blood loss, blood transfusion, extubation time and postoperative shivering. RESULTS: The core temperature at the end of the surgery was (36.4 +/- 0.4) degrees C in the warming group and (35.3 +/- 0.5) degrees C in the control group. The difference was statistically significant (t = 7.547, P < 0.001). There was no significant difference of blood loss and blood transfusion between two groups. The extubation time was significantly shorter in the warming group [(18 +/- 6) vs (26 +/- 10) min, t = -3.364, P = 0.002]. 6 patients shivered postoperatively in the control group and none in the warming group (chi2 = 7.059, P = 0.008). CONCLUSION: Fluid warming system and forced-air warming system can effectively maintain normothermia during the surgery and then help to reduce the extubation time and postoperative shivering.

Abdomen↗

Alterations of adrenomedullin and its receptor system components in calcified vascular smooth muscle cells.

Vascular calcification is a common finding in many cardiovascular diseases. Paracrine/autocrine changes in calcified vessels, and the secreted factors participate in and play an important role in the progress of calcification. Adrenomedullin (ADM) is a potent vasodilator peptide secreted by vascular smooth muscle cells (VSMCs) and vascular endothelial cells. Recently, receptor activity-modifying proteins (RAMPs) have been shown to transport calcitonin receptor-like receptor (CRLR) to the cell surface to present either as CGRP receptor or ADM receptor. In this work, we explored the production of ADM, alterations and significance of ADM mRNA and its receptor system components--CRLR and RAMPs mRNA in calcified VSMCs. Our results showed that calcium content, 45Ca2+ uptake and alkaline phosphatases (ALPs) activity in calcified VSMCs were increased, respectively, compared with control VSMCs. Content of ADM in medium was increased by 99% (p < 0.01). Furthermore, it was found that the levels of ADM, CRLR, RAMP2 and RAMP3 mRNA in calcified cells were elevated, respectively, compared with that of control. The elevated levels of CRLR, RAMP2 and RAMP3 mRNA were significant correlation with ADM mRNA (r = 0.83, 0.92 and 0.93, respectively, all p's < 0.01) in calcified VSMCs. The results show that calcified VSMCs generate an increased amount of ADM, up-regulate gene expressions of ADM and its receptor system components--CRLR, RAMP2 and RAMP3, suggesting an important role of ADM and its receptor system in the regulation of vascular calcification.

Adrenomedullin↗

A caspase-6 and anti-human epidermal growth factor receptor-2 (HER2) antibody chimeric molecule suppresses the growth of HER2-overexpressing tumors.

Clinical studies have suggested that human epidermal growth factor receptor-2 (HER2) provide a useful target for antitumor therapy. We previously described the generation of a chimeric HER2-targeted immunocasp-3 protein. In this study, we extend the repertoire of chimeric proapoptotic proteins with immunocasp-6, a construct that comprises a HER2-specific single-chain Ab, a single-chain Pseudomonas exotoxin A, and an active caspase-6, which can directly cleave lamin A leading to nucleus damage and inducing programmed cell death. We demonstrate that the secreted immunocasp-6 molecule selectively recognizes and induces apoptosis in HER2-overexpressing tumor cells in vitro, but not in cells with undetectable HER2. The immunocasp-6 gene was next transferred into BALB/c athymic mice bearing human breast SK-BR-3 tumors by i.m. injection of liposome-encapsulated vectors, by intratumor injection of adenoviral vectors, or by i.v. injection of PBMC modified by retroviral infection. Regardless of the method used, expression of immunocasp-6 suppressed tumor growth and prolonged animal survival significantly. Our data show that the chimeric immunocasp-6 molecule can recognize HER2-positive tumor cells, promptly attack their nucleus, and induce their apoptotic death, suggesting the potential of this strategy for the treatment of human cancers that overexpress HER2.

ADP Ribose Transferases↗

Patterning of renal cGMP production by the natriuretic peptide receptor type A and blood pressure in spontaneously hypertensive rats.

Although important advances have been made over past decades in studying the mechanisms of hypertension, the nature of cellular signaling patterns involved and their relationship remain unclear. High cGMP production rates in isolated renal glomeruli have been presented as a characteristic of spontaneously hypertensive rat (SHR) even before the development of hypertension, which suggests that this event might be a cause of the increase in blood pressure. Using cross-breeding between SHR and WKY parental strains to obtain F1 and F2 hybrids, we have investigated the patterning of high blood pressure and cGMP production rates. We have found that, in the F2 population, the mean blood pressure and both basal and ANP(1-28)-stimulated cGMP production are similar to the parental SHR. In addition, we have found a positive correlation between blood pressure and high cGMP production rates in the F2 population. The higher cGMP production was not a consequence of hypertension, since in DOCA-salt hypertensive rats cGMP production was similar to that observed in normotensive WKY rats. These observations suggest that high cGMP production is a characteristic linked to hypertension. Finally, reciprocal crosses between the SHR and WKY parental strains showed that in the F1 population blood pressure but not cGMP production are associated with the Y chromosome.

Animals↗

Expression of gonadotropin-releasing hormone receptor and effect of gonadotropin-releasing hormone analogue on proliferation of cultured gastric smooth muscle cells of rats.

AIM: To investigate the expression of gonadotropin-releasing hormone (GnRH) receptor and the effects of GnRH analog (alarelin) on proliferation of cultured gastric smooth muscle cells (GSMC) of rats. METHODS: Immunohistochemical ABC methods and in situ hybridization methods were used to dectect protein and mRNA expression of GnRH receptor in GSMC, respectively. Techniques of cell culture, OD value of MTT test, measure of (3)H-TdR incorporation, average fluorescent values of proliferating cell nuclear antigen (PCNA) and flow cytometric DNA analysis were used in the experiment. RESULTS: The cultured GSMC of rats showed immunoreactivity for GnRH receptor; positive staining was located in cytoplasm. GnRH receptor mRNA hybridized signals were also detected in cytoplasm. When alarelin (10(-9), 10(-7), 10(-5) mol/L) was administered into the medium and incubated for 24 h, OD value of MTT, (3)H-TdR incorporation and average fluorescent values of PCNA all decreased significantly as compared with the control group (P<0.05). The maximum inhibitory effect on cell proliferation was achieved a concentration of 10(-5) mol/L and it acted in a dose-dependent manner. Flow cytometric DNA analysis revealed that alarelin could significantly enhance ratio of G(1) phase and decrease ratio of S phase of GSMC of rats (P<0.05). The maximum inhibitory effect on ratio of S phase was at the concentration of 10(-5) mol/L and also acted in a dose-dependent manner. CONCLUSION: Our data suggest that GnRH receptor can be expressed by GSMC of rats. GnRH analogue can directly inhibit proliferation and DNA synthesis of rat GSMC through GnRH receptors.

Animals↗