BACKGROUNDCalreticulin (CRT) is major Ca 2+ -binding chaperone mainly resident in the endoplasmic reticulum (ER) lumen. Recently, it has been shown that non-ER CRT regulates a wide array of cellular responses. We previously found that CRT was up-regulated during hypoxia/reoxygenation (H/R) and this study was aimed to investigate whether CRT nuclear translocation aggravates ER stress (ERS)-associated apoptosis during H/R injury in neonatal rat cardiomyocytes.

METHODSApoptosis rate and lactate dehydrogenase (LDH) leakage in culture medium were measured as indices of cell injury. Immunofluorescence staining showed the morphological changes of ER and intracellular translocation of CRT. Western blotting or reverse transcription polymerase chain reaction was used to detect the expression of target molecules.

RESULTSCompared with control, H/R increased apoptosis rate and LDH activity. The ER became condensed and bubbled, and CRT translocated to the nucleus. Western blotting showed up-regulation of CRT, Nrf2, activating transcription factor 4 (ATF4), CHOP and caspase-12 expression after H/R. Exogenous CRT overexpression induced by plasmid transfection before H/R increased cell apoptosis, LDH leakage, ER disorder, CRT nuclear translocation and the expression of ERS-associated molecules. However, administration of the ERS inhibitor, taurine, or CRT siRNA alleviated cell injury, ER disorder, and inhibited ERS-associated apoptosis.

CONCLUSIONSOur results indicated that during H/R stress, CRT translocation increases cell apoptosis and LDH leakage, aggravates ER disorder, up-regulates expression of nuclear transcription factors, Nrf2 and ATF4, and activates ERS-associated apoptosis.

Animals ; Apoptosis ; genetics ; physiology ; Calreticulin ; genetics ; metabolism ; Cell Hypoxia ; genetics ; physiology ; Cell Survival ; genetics ; physiology ; Cells, Cultured ; Endoplasmic Reticulum Stress ; physiology ; Myocytes, Cardiac ; cytology ; metabolism ; RNA Interference ; Rats

Objective To evaluate the potential relationship between isoforms of BRCA1 associated RING domain 1 ( BARD1 ) and the pathophysiologic markers of sporadic breast carcinoma of female Han ethnic group. Methods The expression of BARD1 isoforms in 39 breast carcinomatous tissue, 12paracancerous-normal breast tissue and 7 controlled normal breast tissue was detected by reverse transcription polymerase chain reaction (RT-PCR) and then cloned and sequenced.The difference of isoforms expression and their clinical significance were analyzed. Results There were four transcriptive products of BARD1 found in all these candidates,named full lenth,isoform γ,isoform δ and isoform e.The positive rate of isoform γ and δ was higher in carcinomatous tissues than in paracancerous-normal tissues and normal breast tissue in healthy women ( P ＜ 0.05 ). Carcinomatous tissue expressed more kinds of isoforms.There was significant difference between carcinomatous tissue and paranormal/nomal tissue ( P =0.0075 ).There was significant correlation between isoform ε positive and poor prognosis factors such as poorly differentiation,HER2 positive,poor pathologic type and larger breast cancer lumps(P ＜ 0.05 ). Conclusions There are significant differences in the expression of BARD1 isoforms among different kinds breast tissues in the female Han ethnic group.Positive isoform ε may predict poor prognosis of breast carcinoma in the female Han ethnic group.

Objective To explore the effect of different indexes on evaluating insulin resistance in obese children. Methods Oral glucose tolerance test (OGTT) was performed in obese children (n= 61) and age - matched normal volunteers( n= 23) Serum glucose and insulin levels were determined at 0,30,60,120,180 min after OGTT, insulin resistance was estimated by homeostasis model assessment of insulin resistance (HOMA-IR). Indexes such as the ratio of area under the curve of glucose(AUCG)/area under the curve of insulin(AUCI), the ratio of fasting blood sugar(FBG) and fasting blood insulin (FINS) were meanwhile calculated. Results The level of serum FINS was significantly higher in obese children(P

OBJECTIVETo investigate the characteristics, diagnosis, and treatment of ovotesticular disorder of sex development (OT-DSD).

METHODSWe retrospectively analyzed 2 cases of OT-DSD treated in our hospital. The patients were 19 and 15 years old, respectively, and both received systematic physical examination and examinations of the karyotype, sex hormone, adrenocorticotropic hormone (ACTH), color Doppler ultrasonography, urethrocystoscopy, and human chorionic gonadotropin (HCG) test. Under the laparoscope, we performed surgical gonad exploration, gonadectomy, and vulvar orthopedics. Intraoperative exploration and pathology confirmed true hermaphroditism in both cases, with sex selection as female. One underwent laparoscopic resection of the ovotestis, and the other removal of the testis with the ovarian tissue reserved.

RESULTSThe patients were followed up for 12 months postoperatively, which found no abnormality in either the vulvas or the genital glands.

CONCLUSIONSurgical exploration of the gonad is the only method for the diagnosis of OT-DSD and sex selection is the key to treatment. Laparoscopic surgical exploration of the gonad and vulvar orthopedics are the first treatment options.

Adolescent ; Chorionic Gonadotropin ; Female ; Gonadal Steroid Hormones ; Humans ; Karyotype ; Laparoscopy ; Male ; Ovary ; Ovotesticular Disorders of Sex Development ; diagnosis ; surgery ; Retrospective Studies ; Testis ; surgery ; Young Adult

In order to investigate the influencing factors of 5-hydroxymethyl-2-furaldehyde (5-HMF) content in Schisandra, confirm the theory of 5-HMF deriving mainly from Schisandra processing course, and give some suggestions about the Schisandra processing method, the 5-HMF contents in decoctions of Schisandra under different heating temperature, decocting time, soaking time, processing methods and treatment with different solvents before decocting the Schisandra were measured by RP-HPLC method. The results showed that there is great difference of 5-HMF level in decoctions from differently processed Schisandra and unprocessed Schisandra; decocting time of 60 min has some effects on 5-HMF level in decoctions and there is certain quantity 5-HMF in processed Schisandra itself and very little 5-HMF in unprocessed Schisandra. Heating time, heating temperature and treating solvents all have effect on 5-HMF level in decoction of Schisandra. 5-HMF in Schisandra was mainly from processing course. Both long heating time and high heating temperature can increase 5-HMF level in Schisandra. The production of 5-HMF in Schisandra may have some relationships with some polar components, which can dissolve in water, ethanol and acetone, especially in ethanol. To control processing temperature, processing time and treatment with some solvent is very important for controlling 5-HMF level in Schisandra.
Chromatography, High Pressure Liquid ; Furaldehyde ; analogs & derivatives ; analysis ; Plant Extracts ; chemistry ; Schisandra ; chemistry ; Temperature ; Time Factors

Objective To investigate the clinical and mammographic features of plasma cell mastitis.Methods Twenty-five patients(28 lesions)with histologically confirmed plasma cell mastitis, aged from 26 to 70 years(mean age 41 years),were examined with X-ray mammography.The clinical manifestations and imaging features were retrospectively reviewed.Results No case was in lactation.The painful irregular masses,ranged from 1.3 to 8cm in size,were found in 22 patients,while 3 patients with acute episode.Recurrent episodes of breast masses were noted in 4 patients.Based on the mammographic appearances,the plasma cell mastitis were classified as the following four types:inflammation-like type (2/28),ductal ectasia type(3/28),focal infiltration type(10/28)and nodular type(13/28).The valuable radiogyaphic signs:(1)An asymmetrically increased density along the lactiferous duct with a flame-like appearance,inhomogeneous low density tubular structures and scattered stick-shape calcifications.(2) Architectural distortion and oil cysts formation in adjacent area,(3)Subareolar ductal ectasia.Conclusions The clinical and mammographic characteristics of plasma cell mastitis are critical to avoiding unnecessary surgery.Histopathological result is needed for the diagnosis in patients highly suspected of malignancy.

OBJECTIVETo study changes of left ventricular remodeling (LVR) in hypertension patients with carotid atherosclerosis (CAS) of phlegm-dampness syndrome (PDS).

METHODSDoppler ultrasonography data of CAS were observed in 223 hypertension patients with CAS (as the hypertension group, including 119 patients of the PDS group and 104 of the non-PDS group), 81 CAS patients with non-hypertension, and 19 non-hypertension non-CAS patients (as the control group). The difference in the degree of LVR was compared among the above groups.

RESULTSThe left ventricular posterior wall thickness (LVPWT), inter ventricular septum thickness (IVS), E/A were higher in the hypertension group than in the non-hypertension group (P < 0.05). The left ventricular end-diastolic dimension (LVEDD), left ventricular end-systolic diameter (LVESD), stroke volume (SV) were higher in the soft plaque hypertension group and the soft plaque non-hypertension group than in the hard plaque group, the thickening intimal group, and the normal intimal group (P < 0.01 , P < 0.05). The LVEDD, LVESD, and SV were higher, and the ejection fraction (EF) was lower in the PDS hypertension group than in the non-PDS hypertension group (all P < 0.05). Of them, LVEDD, LVESD, and SV were higher in the soft plaque group than in the hard plaque group (P < 0.01), the thickening intimal group (P < 0.01) and the normal intimal group (P < 0.05). There was no statistical difference in PDS hypertension between the soft plaque group and the hard plaque group (P > 0.05).

CONCLUSIONThe hypertension patients with CAS of PDS might be correlated to LVR, and LVR was more obviously in the soft plaque patients.

Aged ; Aged, 80 and over ; Carotid Artery Diseases ; diagnosis ; diagnostic imaging ; physiopathology ; Case-Control Studies ; Female ; Humans ; Hypertension ; diagnosis ; diagnostic imaging ; physiopathology ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Ultrasonography ; Ventricular Remodeling

Objective To compare the antimierobial resistance of the gram-negative isolates from blood stream between Beijing and Hong Kong patients.Methods Non duplicate blood stream isolates were collected between 1998 and 2003 from Peking U- nion Medical College Hospital(n=530)and Hong Kong Prince of Wales Hospital(n=2913).Disk diffusion method and broth microdilution(MB)method were used to determine the antimierobial susceptibility of these strains to 13 antimicrobial a- gents including imipenem,ceftazidime,cefepime,etc.Results The resistance rates of E.coli to amikaein,ceftazidime and cefepime were 0.6%-5.0% and 7.5%-23.5%,and to gentamicin,cefuroxime,ciprofloxacin and cefuroxime were 12.6%- 28.4% and 29.4%-68.4% in Hong Kong isolates(n=1471)and Beijing isolates(n=213),respectively.The resistance rates of Klebsiella spp.to piperacillin-tazobactam and ceftazidime were 3.3%-11.3% and 4.3%-17.7%,and to amikacin, gentamicin,cefotaxime and ceftriaxone were 1.5%-10.1% and 5.8%-27.1% in Hong Kong isolates(n=586)and Beijing i- solates(n=70),respectively.The strains above and the ECS group of bacteria in Hong Kong(consisting of Enterobacter spp.,Citrobacter spp.,and Serratia spp.,n=333)were all susceptible to imipenem and meropenem,but 4.6%(n=65)of the ECS group bacteria in Beijing were resistant.About 8.8% and 14% of ECS group isolates were resistant to cefepime.Such isolates resistant to the other?-lactams ranged from 28.4% to 96%.In Hong Kong,Salmonella typhi remained susceptible to most of the antimierobial agents tested,but in Beijing,the resistance rates to ampicillin and piperacillin were from 9.1% to 18.2%.In Hong Kong,The resistance rates of Acinetobacter spp.and P.aeruginosa to imipenem and meropenem were lower than 10%,but in Beijing their resistance rates to imipenem were 13.6%-15.3% and 8.8%-30.8%,respectively.Conclusions The resistance rates of gram-negative bacteria isolated from blood stream in Beijing to most antimicrobial agents were higher than the corresponding rates in Hong Kong.This result will be useful for empirical therapy of local bacteremia.

The present study was aimed to investigate whether calreticulin (CRT) was involved in the protective effect of hypoxic preconditioning (HPC) against oxidative stress injury in rat cardiomyocytes. Neonatal cardiomyocytes were prepared from Sprague-Dawley rats aged 24 h, and cultured in DMEM medium containing 10% fetal bovine serum. The cultured cardiomyocytes were randomly divided into 8 groups as follows: (1) hydrogen peroxide stress (H(2)O(2) group); (2) brief hypoxic exposure for 20 min to simulate HPC (HPC group); (3) hypoxic exposure for 20 min followed by normoxic reoxygenation for 24 h before hydrogen peroxide stress (HPC + H(2)O(2) group); (4) SB203580 (a specific inhibitor of p38 MAPK) + HPC + H(2)O(2) group; (5) CRT antisense oligonucleotide transfection (AS group); (6) AS + H(2)O(2) group; (7) AS + HPC + H(2)O(2) group; (8) control group. Morphological observation, lactate dehydrogenase (LDH) leakage and flow cytometry were employed to assess cell apoptosis and necrosis. RT-PCR and Western blot were used to detect CRT expression and activity of p38 MAPK. All experiments were repeated at least four separate times. The results obtained were as follows: (1) HPC relieved cell injury caused by H(2)O(2). Compared with that in H(2)O(2) group, the cell survival rate increased by 18.0% (P<0.05), apoptotic rate and LDH leakage in culture medium decreased by 19.4% and 53.0%, respectively (P<0.05) in HPC + H(2)O(2) group. (2) H(2)O(2) induced CRT over-expression (7.1-fold increase compared with control, P<0.05), while HPC resulted in mild CRT up-regulation (2.4-fold increase compared with control, P<0.05), suggesting that HPC can relieve the over-expression of CRT induced by H(2)O(2). (3) CRT AS transfection weakened the protection of HPC. Compared with that in HPC + H(2)O(2) group, the cell survival rate decreased by 4% (P<0.05), and apoptotic rate and LDH leakage in culture medium increased by 2.6% and 39.0%, respectively (P< 0.05) in AS + HPC + H(2)O(2) group. (4) The protection of HPC and HPC-induced upregulation of CRT were almost eliminated when SB203580 was administered before HPC. These results suggest that HPC up-regulates CRT expression through the p38 MAPK signaling pathway and protects cardiomyocytes from oxidative stress injury.
Animals ; Apoptosis ; Calreticulin ; metabolism ; Cell Hypoxia ; Cell Survival ; Cells, Cultured ; Hydrogen Peroxide ; Imidazoles ; pharmacology ; Ischemic Preconditioning, Myocardial ; L-Lactate Dehydrogenase ; metabolism ; Myocytes, Cardiac ; pathology ; Oxidative Stress ; Pyridines ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Up-Regulation ; p38 Mitogen-Activated Protein Kinases ; antagonists & inhibitors

In order to understand the intracellular mechanism of preconditioning, we investigated the relationship among activities of extracellular signal-regulated protein kinases (ERKs), the expression of hypoxia-inducible factor -1alpha (HIF-1alpha) and the effect of hypoxic preconditioning (HPC) on cell injury induced by hypoxia-reoxygenation in cultured neonatal rat cardiomyocytes 24 h after brief hypoxia. Cultured cardiomyocytes of neonatal Sprague-Dawley rats were divided into four groups: hypoxia/reoxygenation (H/R), hypoxia preconditioning (HPC), hypoxia preconditioning + mitogen-activated protein kinase (MAPK) inhibitor PD98059 (HPC+PD98059), and control (C). We measured the survival rate and apoptosis rate of cardiomyocytes at 6 or 12 h after hypoxia/reoxygenation, activities of extracellular signal-regulated protein kinases (ERKs), and expression of hypoxia-inducible factor-1alpha (HIF-1alpha). We found that the survival rate of cardiomyocytes in hypoxic preconditioning group increased by 6.08% and 7.91% at 6 and 12 h after hypoxia/reoxygenation (n=6, P<0.05), respectively, and the apoptotic rate decreased by 10.92% and 14.34% (n=6, P<0.05) respectively. Hypoxic preconditioning increased the abundance of phospho-ERK1/2 by 3-folds and expression of HIF-1alpha by 1-fold in whole cell extracts from hypoxic preconditioned cardiomyocytes. PD98059, an inhibitor of the upstream kinase of ERKs, abolished the anti-injury effect, ERKs activation, and expression of HIF-1alpha induced by hypoxic preconditioning. Statistical analysis indicated that there was negative correlation between apoptotic rate and activities of ERKs or expression of HIF-1alpha, and positive correlation between activities of ERKs and expression of HIF-1alpha. It is concluded that hypoxic preconditioning protects cardiomyocytes from hypoxia/reoxygenation-induced injury and that upregulation of HIF-1alpha through ERKs pathway mediates the cardioprotection of hypoxic preconditioning.
Animals ; Animals, Newborn ; Apoptosis ; drug effects ; Cell Hypoxia ; Cell Survival ; Cells, Cultured ; DNA-Binding Proteins ; biosynthesis ; physiology ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Hypoxia-Inducible Factor 1 ; Hypoxia-Inducible Factor 1, alpha Subunit ; Ischemic Preconditioning, Myocardial ; Myocytes, Cardiac ; cytology ; pathology ; Nuclear Proteins ; biosynthesis ; physiology ; Rats ; Rats, Sprague-Dawley ; Transcription Factors ; biosynthesis ; physiology