OBJECTIVETo explore cell death and apoptosis in rat hippocampal neurons at different time points after ischemia, hypoxia and reperfusion injury and to elucidate time window characteristics in ischemia neuronal injury.

METHODSHippocampal neurons were obtained from rat embryo and were cultured in vitro. The ischemia and reperfusion of cultured rat hippocampal neurons were simulated by oxygen-glucose deprivation (OGD) and recovery. OGD at different time points (0.25 h to 3.0 h) and then the same recovery (24 h) were prepared. Annexin V-PI staining and flow cytometry examined neuron death and apoptosis at different time after injury.

RESULTSAfter OGD and recovery, both necrosis and apoptosis were observed. At different times after OGD, there were statistically significant differences in neuron necrosis rate (P < 0.05), but not in apoptosis rate (P > 0.05). At recovery, survival rate of hippocampal neurons further decreased while apoptosis rate increased. Furthermore, apoptosis rates of different time differed greatly (P < 0.05). Apoptosis rate gradually increased with significant difference among those of different time points (P < 0.05). However, 2 h after ischemia, apoptosis rate decreased markedly.

CONCLUSIONSApoptosis is an important pathway of delayed neuron death. The therapeutic time window should be within 2 h after cerebral ischemia and hypoxia.

Animals ; Apoptosis ; physiology ; Brain Ischemia ; pathology ; Cell Death ; physiology ; Cell Hypoxia ; Cells, Cultured ; Disease Models, Animal ; Female ; Fetus ; cytology ; Flow Cytometry ; Hippocampus ; pathology ; Neurons ; pathology ; Pregnancy ; Pregnancy, Animal ; Probability ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; pathology ; Sensitivity and Specificity ; Time Factors

OBJECTIVETo prepare cryptotanshinone (CT)-cyclodextrin inclusion compound and improve dissolution of CT.

METHODInclusion ratio was determined by plotting the phase solubility curve of CT versus hydroxypropyl-beta-cyclodextrin (HPCD). CT-cyclodextrin inclusion compound was made by wet grinding method. Properties of the inclusion compound was investigated by in vitro dissolution test, DTA and IR spectrum.

RESULTInclusion ratio of CT versus HPCD was 1:1. Dissolution of CT-HPCD inclusion compound at 45 min was 21.6 times of material drug.

CONCLUSIONDissolution of CT was improved remarkably in CT-HPCD inclusion compound. The complexation force of the inclusion compound was hydrogen bond formed by carbonyl group of CT and hydroxyl group of HPCD.

2-Hydroxypropyl-beta-cyclodextrin ; Biological Availability ; Drug Carriers ; Drugs, Chinese Herbal ; chemistry ; Phenanthrenes ; chemistry ; isolation & purification ; Salvia miltiorrhiza ; chemistry ; Solubility ; Technology, Pharmaceutical ; methods ; Time Factors ; beta-Cyclodextrins ; chemistry

Adolescent ; Antibodies, Viral ; blood ; China ; epidemiology ; Female ; Humans ; Influenza A virus ; immunology ; Male ; Seroepidemiologic Studies ; Students, Medical ; Vietnam ; epidemiology ; Young Adult

OBJECTIVETo develop an HPLC method for determining oxyphyllenodiol A (1) and teuhetenone A (2) contained in Alpinia oxyphylla and to compare the contents of the two components contained in medicinal materials and prepared herbal medicines in pieces sold in the market and different fractions.

METHODHPLC and Waters sunfire C18 column (4. 6 mm x 250 mm, 5 microm) were adopted for gradient elute with the mobile phase of acetonitrile and water. The flow rate was 1.0 mL x min(-1) and the detection wavelength was 250 nm.

RESULT1 and 2 showed a good linear relationship within the range of 0.1296 - 0.8640 microg and 0.1635 - 1.0900 microg respectively, with the average recoveries of 99.08% and 97.80%. Their content ranges were 0.0059% - 0.0149% and 0.0080% - 0.0164% in different samples. The mean value of 1 and 2 were 0.0085% and 0.0104% in the whole fruits, and 0.0137% and 0.0157% in the seeds. They were undetected in the nutshells.

CONCLUSIONThe method is so precise, accurate and highly reproducible that it can be used to determine the contents of oxyphyllenodiol A and teuhetenone A in A. oxyphylla. The contents of the two components are mainly extracted from the seeds, with certain difference among different samples. There are a higher contents and no significant difference in the salted and raw seeds.

Alpinia ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; isolation & purification ; Sesquiterpenes ; analysis ; isolation & purification

Objective To evaluate the efficacy of a novel hemostatic material AristaTM in the management of active and local bleeding in neurosurgery, and discuss the indications for its application. Methods Forty-eight patients undergoing elective craniotomy in our department between April, 2008 and May, 2009 were randomized into the test group (n=24) and control group (n=24) with intraoperative hemostatic management using AristaTM and gelatin sponge, respectively. The hemostasis time and efficacy of the two materials were compared. Results Hemostasis was effective in all the 48 patients. The mean hemostasis time in the test group and control group was 1.88±0.74 min and 3.38±0.92 min, respectively, showing a significant difference between them (Z=4.711, P=0.001). Conclusion AristaTM allows more efficient management of active and local bleeding than gelatin sponge during neurosurgeries and has great potential for clinical application.

AIM:To elucidate the association between chronic kidney injury and interleukin-33 (IL-33;an alarmin)/suppression of tumorigencity 2 (ST2) in patients with systemic lupus erythematosus (SLE).METHODS:Serum levels of IL-33 and soluble ST2 (sST2) were assessed by ELISA in 50 SLE patients and 30 healthy controls (HC).RESULTS:The levels of IL-33 and sST2,and IL-33/sST2 ratio were significantly higher in SLE patients than those in the HC.The IL-33 and sST2 levels were positively associated with SLE disease activity index (SLEDAI),erythrocyte sedimentation rate (ESR),proteinuria and triglyceride,but negatively associated with complement C3.IL-33/sST2 ratio was positively associated with SLEDAI and estimated glomerular filtration rate (eGFR).Independent explanatory variables associated with high IL-33/sST2 included chronic kidney disease (CKD) staging and albumin (R2 =0.442),especially CKD staging.CONCLUSION:Elevated serum sST2 and IL-33 levels in SLE patients are correlated with disease activity and risk factors of kidney injury.IL-33/sST2 ratio may serve as a potential biomarker for chronic kidney injury in SLE patients.

OBJECTIVEto investigate the effect of somatostatin on inflammatory immune disorders and prognosis in patients with severe sepsis caused by abdominal diseases.

METHODSfifty-three patients with severe abdominal sepsis (age > 18 years, APACHE-II score > 15) from June 2005 to June 2009 were randomly divided into Somatostatin group (n = 23) and SSC Group (n = 30). Fifteen healthy volunteers of the same age range were chosen as Control group. The SSC group was treated with classical SSC therapy, and the Somatostatin Group was treated with the same regime plus 14-peptide somatostatin continuous infusion at the dose of 6 mg/24 h for 7 days. The serum levels of interleukin-10 (IL-10), tumor necrosis factor-α (TNF-α) were determined by using ELISA. CD(4)(+), CD(8)(+) T cell subsets were determined by fluorescence activated cell sorter(FACS) and CD(4)(+)/CD(8)(+) was calculated. APACHE-II score was observed on admission (d1) and day 3, 7 and 14 after treatment. Morality rates in 28 days in two groups were recorded.

RESULTScompared with Control group, IL-10 and TNF-α levels were significantly elevated in patients with severe abdominal sepsis (P < 0.05), while CD(4)(+), CD(8)(+) T cell and CD(4)(+)/CD(8)(+) decreased significantly (P < 0.05). Compared with the Somatostatin group CD(4)(+), CD(8)(+) T cell and CD(4)(+)/CD(8)(+) on d7 and d14 in SSC Group were significantly increased (P < 0.05), while IL-10 and TNF-α decreased significantly(P < 0.05). APACHE-II scores on d3, d7, d14 of Somatostatin group were significantly lower than those of SSC group, and 28 d mortality rate also declined.

CONCLUSIONSin patients with severe abdominal sepsis, systemic inflammatory response and immune suppression exist simultaneously. Somatostatin has a dual immunomodulatory activity in these patients.

APACHE ; Case-Control Studies ; Female ; Humans ; Interleukin-10 ; blood ; Male ; Prognosis ; Prospective Studies ; Sepsis ; drug therapy ; etiology ; immunology ; Somatostatin ; therapeutic use ; T-Lymphocyte Subsets ; immunology ; Tumor Necrosis Factor-alpha ; blood

OBJECTIVECarnitine deficiency has been associated with progressive cardiomyopathy due to compromised energy metabolism. The objective of this study was to investigate clinical features of carnitine deficiency-induced cardiomyopathy and the therapeutic efficacy of L-carnitine administration.

METHODBetween January 2010 and December 2011, filter-paper blood spots were collected from 75 children with cardiomyopathy. Free carnitine and acylcarnitine profiles were measured for each individual by tandem mass spectrometry (MS/MS). For those in whom carnitine deficiency was demonstrated, treatment was begun with L-carnitine at a dose of 150 - 250 mg/(kg·d). Clinical evaluation, including physical examination, electrocardiography, chest x-ray, echocardiography and tandem mass spectrometry, was performed before therapy and during follow-up.

RESULTOf 75 cardiomyopathy patients, the diagnosis of carnitine deficiency was confirmed in 6 patients, which included 1 boy and 5 girls. Their age ranged from 0.75 to 6 years. Free carnitine content was (1.55 ± 0.61) µmol/L (reference range 10 - 60 µmol/L). Left ventricular end-diastolic diameter (LVDd) was (5.04 ± 0.66) cm and left ventricular ejection fraction (LVEF) was (38.5 ± 10.5)%. After 10 - 30 d therapy of L-carnitine, free carnitine content rose to (30.59 ± 15.02) µmol/L (t = 4.79, P < 0.01). LVDd decreased to (4.42 ± 0.67) cm (t = 4.28, P < 0.01) and LVEF increased to (49.1 ± 7.6)% (t = 6.59, P < 0.01). All patients received follow-up evaluations beyond 6 months of treatment. Clinical improvement was dramatic. LVEF returned to normal completely in all the 6 patients. LVDd decreased further in all the 6 patients and returned to normal levels in 3 patients. No clinical signs or symptoms were present in any of the 6 patients. The only complications of therapy had been intermittent diarrhea in 1 patient.

CONCLUSIONTandem mass spectrometry is helpful to diagnose carnitine deficiency and should be performed in all children with cardiomyopathy. L-carnitine has a good therapeutic effect on carnitine deficiency-induced cardiomyopathy.

Adolescent ; Cardiomyopathies ; diagnosis ; drug therapy ; etiology ; Cardiotonic Agents ; administration & dosage ; therapeutic use ; Carnitine ; blood ; deficiency ; therapeutic use ; Child ; Child, Preschool ; Electrocardiography ; Female ; Follow-Up Studies ; Humans ; Infant ; Male ; Retrospective Studies ; Tandem Mass Spectrometry ; Treatment Outcome ; Ventricular Function, Left ; drug effects

OBJECTIVETo identify the virus isolated from Jiangmen, Guangdong province and to discuss the possible origin.

METHODSUsing characteristics of indirect fluorescent antibody tests (IFA), reverse transcription-polymerase chain reaction (RT-PCR), mouse neurovirulence and cell culture to identify the isolated virus. According to the nature of dengue virus type 2 NGC strain, two pairs of primers were designed. The structural protein gene of isolated dengue virus type 2 strain was then amplified by RT-PCR, cloned into pMD18-T vector and sequenced.

RESULTSTwenty-two of 37 serum samples showed a positive reaction to dengue antibody IgG, and 36 of 37 with IgM with the highest antibody titer 1:640. Ten samples were resulted in a cytopathy on C6/36 cells and showed a neurovirulence in suckling mice when inoculated intracerebrally. The structural gene of new isolate GD19/2001 containing 2 325 nucleotides which encoded 774 amino acids. Data on nucleotide homology were 98%, 96%, 94%, 94%, 92%, 92%, 92% and 91% compared with TSV01, GD06/93, NGC and 44, ThNH81/93, 04 and GD08/98, and S1 respectively.

CONCLUSIONThe isolated virus from Jiangmen, Guangdong province belonged to dengue virus type 2, which might come from Australia.

Animals ; Antibodies, Viral ; blood ; China ; epidemiology ; DNA, Viral ; genetics ; Dengue ; epidemiology ; virology ; Dengue Virus ; genetics ; immunology ; isolation & purification ; Fluorescent Antibody Technique ; Humans ; Polymerase Chain Reaction ; RNA, Viral ; genetics ; Sequence Analysis, DNA

OBJECTIVETo investigate the feasibility and clinical significance of detecting sperm mitochondrial function by using Rh123/PI dual fluorescent staining and flow cytometry analysis, and to explore the relationship between the results of Rh123/PI dual fluorescent staining and seminal parameters.

METHODSSixty-three semen samples were classified as normal (n=31) and abnormal (n=32) according to the World Health Organization guidelines. Rh123/PI dual fluorescent staining was then carried out to evaluate sperm mitochondrial function by flow cytometry analysis.

RESULTSSignificant differences in Rh123+ PI-, Rh123- /PI+ and Rh123- /PI- were detected between the normal and abnormal semen samples (P < 0.05). There was a significant positive correlation between the Rh123+ PI- sperm and sperm motility and a significant inverse correlation between Rh123+ PI- and immotile sperm. But the Rh123- PI+ sperm showed a contrary relationship with Rh123+ PI-. A significant inverse correlation was also observed between the Rhl23- /PI- sperm and sperm concentration in the abnormal group.

CONCLUSIONRh123/PI dual fluorescent staining and flow cytometry analysis can readily and quickly detect sperm mitochondrial function and be used to evaluate semen quality.

Adult ; Case-Control Studies ; Flow Cytometry ; Humans ; Male ; Mitochondria ; physiology ; Rhodamine 123 ; Sperm Count ; Sperm Motility ; Spermatozoa ; Staining and Labeling ; methods