Whey protein concentrate (WPC 80) and sodium caseinate were hydrolyzed by Protamex to 5%, 10%, 15%, and 20% degree of hydrolysis (DH). WPC 80, sodium caseinate and their hydrolysates were then analyzed, compared and evaluated for their nutritional qualities. Their chemical composition, protein solubility, amino acid composition, essential amino acid index (EAA index), biological value (BV), nutritional index (NI), chemical score, enzymic protein efficiency ratio (E-PER) and in vitro protein digestibility (IVPD) were determined. The results indicated that the enzymatic hydrolysis of WPC 80 and sodium caseinate by Protamex improved the solubility and IVPD of their hydrolysates. WPC 80, sodium caseinate and their hydrolysates were high-quality proteins and had a surplus of essential amino acids compared with the FAO/WHO/UNU (1985) reference standard. The nutritive value of WPC 80 and its hydrolysates was superior to that of sodium caseinate and its hydrolysates as indicated by some nutritional parameters such as the amino acid composition, chemical score, EAA index and predicted BV. However, the E-PER was lower for the WPC hydrolysates as compared to unhydrolyzed WPC 80 but sodium caseinate and its hydrolysates did not differ significantly. The nutritional qualities of WPC 80, sodium caseinate and their hydrolysates were good and make them appropriate for food formulations or as nutritional supplements.
Amino Acids ; chemistry ; Caseins ; chemistry ; metabolism ; Dietary Proteins ; analysis ; Evaluation Studies as Topic ; Hydrolysis ; Milk Proteins ; chemistry ; metabolism ; Models, Statistical ; Nutritive Value ; Protein Hydrolysates ; chemistry ; Solubility ; Temperature ; Time Factors ; Tryptophan ; chemistry ; Whey Proteins

Objective To study the neuroprotective effect of erythropoietin(EPO) on neonatal rats model with hypoxia-ischemia encephalopathy(HIE).Methods HIE was induced in rats on 7th day of postnatal age by ligation of right common carotid artery,followed by 2 h of hypoxia(80 mL/L O2).The subjects were divided into sham-operated group,control group and EPO group.EPO 4 000 U/(kg?day) was injected daily from day 2 pre-surgery for 9 to 16 days and PBS was injected in the control group.The neuroprotective effect of EPO on HIE model was detected by brain weight,the difference in weights between the ipsilateral(right) and contralateral(left) brain and the function test.In vitro study,the neural progenitor cell line C17.2 under gone apoptosis following an ischemia-like metabolic inhibition.The effect of EPO on the cell line ischemia modle 17.2 was evaluated by detecting Annexin V with flow cytometry.Results The signi-ficant and sustained brain injury in the hypoxia-ischemia and vehicle-treated group was observed and measured by reduction in relative weights of ipsilateral to contralateral and compromised sensorimotor functions in response to postural reflex test,compared with those of sham-operated animals(Pa

Objective To discuss a real-time fluorescence quantitative polymerase chain reaction (FQ-PCR) wether if can be used to detect Brucella. Methods According to the BCSP31 gene sequences specific for Brucella, one pair of primers and one TaqMan probe were designed. A real-time PCR was developed with the BCSP31 fragments cloned into PMD18-T vector. The standard cure was established and the sensitivity, the species specificity and the stability of the assay were evaluated. The clinical blood specimens were detected by QT-PCR and compared with clinical diagnosis. Results The standard curve was established with the standard template and the relationship between the Ct and the DNA copy number was linear(r=0.999). The sensitivity of the real-time PCR was 5 copies/μl. The sensitivity of the common PCR was 5×102 copies/μl. The sensitivity was about 100 times higher than common PCR. Species specificity of this FQ-PCR assay evaluated using genomic DNA from 6 Bmcella strains and 5 non-Brucella strains and strong fluorescence was detected in all Brucella strains. The CV of intra-assay and inter-assay reproducibility were 0.71%,7.23%, reprectively. Twenty-four specimens from clinical brucellosis cases, 19 showed positive, the positive coincident rate was 79%(19/24). The negative results were obtained for all 31 negative control, and the negative coincident rate was 100%(31/31). Two were positive from all 30 specimens clinically suspected. Conclusions Highly specific, sensitive, repeatable and coincidental with clinic, this FQ-PCR is quite useful for rapid detection of tiny DNA of Brucella in various samples and laboratory diagnosis.

Adult ; Female ; Gastrointestinal Hemorrhage ; metabolism ; Glutathione ; therapeutic use ; Glutathione Peroxidase ; metabolism ; Humans ; Lipid Peroxidation ; Liver ; blood supply ; Liver Cirrhosis ; complications ; metabolism ; Male ; Middle Aged ; Reperfusion Injury ; prevention & control

OBJECTIVEThe cascade of physiological events underlying hypoxic-ischemic brain damage (HIBD) remains to be fully established. The perinatal brain shows both an increased tolerance to hypoxic-ischemic (HI) injury and a faster and more complete recovery than the adult. It is, therefore, important to understand the sequence of events following hypoxia and ischemia in young animals. The present study aimed to clarify the time-course of the activation of the mu-calpain, and the expression of c-Fos, c-Jun, HSP70 and HSP27 proteins following severe HI (2 h hypoxia) and their relationship with each other.

METHODSA modified newborn rat model of HIBD that included a combination of hypoxia and ischemia as described by Rice was used. Forty-two postnatal 7-day-old Sprague-Dawley rats were randomly divided into seven groups (6 rats in each): 6 time-window groups and a normal control group. Samples were collected at 0, 1, 2, 4, 12 and 24 h after HI insults. The protein concentration was determined using a modified Bradford assay. mu-calpain activation, c-Fos, c-Jun, HSP70 and HSP27 expressions were observed respectively by Western blot from cortical and hippocampal samples.

RESULTSThe cleavage of cytosolic mu-calpain was observed from both cortical and hippocampal samples in neonatal rats after HI. The ratio 76:80 of mu-calpain was increased significantly post-HI and reached a maximum at 24 h in cortex and at 12 h in hippocampus after HI. The expressions of c-Fos and c-Jun from both cortical and hippocampal samples in neonatal rats were up-regulated and peaked at 2 or 4 h after HI, demonstrating significant differences at 1, 2, 4, and 12 h compared with that observed in the control (P < 0.05). When compared with that observed in cortex, the nuclear c-Fos expression from hippocampal samples was highly elevated at 2, 4 and 12 h but significantly decreased at 24 h after HI (P < 0.05), while the nuclear c-Jun expression from hippocampal samples was highly elevated at 0 and 1 h but significantly decreased at 4 and 24 h after HI (P < 0.05). Similarly, the expressions of HSP70 and HSP27 from both cortical and hippocampal samples were up-regulated and reached a maximum at 12 or 24 h after HI, demonstrating significant differences at 12 or 24 h both in cortex and hippocampus for HSP70, and at 24 h in cerebral cortex as well as at 12 and 24 h in hippocampus for HSP27 compared with the control (P < 0.05). Furthermore, in comparison with that observed in cortex, the HSP70 expression from hippocampal samples was highly elevated at 1 h, but significantly decreased at 4, 12 and 24 h after HI (P < 0.05), while the HSP27 expression was permanently elevated in hippocampus after HI.

CONCLUSIONThe neuronal injury induced by HI insults appears to involve many ongoing and simultaneous mechanisms. HI activates the calpains immediately, which may contribute to neuron apoptosis, and induces a significant brain neuroprotection, since there is an increased HSP70 expression and a relatively late remarkable HSP27 expression in hypoxic-ischemic neonatal rat brain. Nuclear c-Fos and c-Jun may participate in the pathogenesis of HIBD.

Animals ; Animals, Newborn ; Blotting, Western ; Brain ; metabolism ; pathology ; Calpain ; metabolism ; Enzyme Activation ; Female ; HSP27 Heat-Shock Proteins ; HSP70 Heat-Shock Proteins ; metabolism ; Heat-Shock Proteins ; metabolism ; Hypoxia, Brain ; metabolism ; Male ; Neoplasm Proteins ; metabolism ; Proteins ; metabolism ; Proto-Oncogene Proteins c-fos ; metabolism ; Proto-Oncogene Proteins c-jun ; metabolism ; Rats ; Rats, Sprague-Dawley ; Time Factors

Objective To study the application value of dynamic monitoring of blood pressure in the prevention and treatment of elderly hypertension. Methods 519 hypertensive patients from December 2017 to December 2018 were monitored with ambulatory blood pressure monitoring, and were divided into the elderly group (≥60 years old, 264 cases) and the control group (<60 years old, 255 cases). The results of ambulatory blood pressure monitoring in two groups were analyzed, which inclued the circadian rhythm of blood pressure, blood pressure, pulse pressure, coefficient of variation of blood pressure, blood pressure load value, average heart rate and morning blood pressure surge. Results The incidence of abnormal circadian rhythm of ambulatory blood pressure in the elderly group was 76.5%. Compared with the control group, there were differences in the indexes of diastolic blood pressure (DBP), diastolic blood pressure load value (DBPLV), pulse pressure (PP), 24 h average heart rate (24 hAHR), systolic blood pressure coefficient of variation (SBPCV), 24 h diastolic blood pressure coefficient of variation (24 h DBPCV) and morning diastolic blood pressure surge (MDBPS) between the two groups(all P<0.05). There were differences in 24 h systolic blood pressure (24hSBP), night systolic blood pressure (nSBP), night diastolic blood pressure (nDBP), night pulse pressure (nPP), day systolic blood pressure load value (dSBPLV), ninght systolic blood pressure load value (nSBPLV), 24 h SBPCV, 24 h dDBPCV and other indicators among different blood pressure types in the elderly group (all P<0.05). Conclusion Ambulatory blood pressure monitoring indicators have important guiding value for the prevention and treatment of elderly hypertension.

OBJECTIVETo investigate the effect of human platelet lysates (HPL) obtained from platelet-rich plasma on the proliferation and biological characteristics of human mesenchymal stem cells (MSCs) in vitro.

METHODSHPL was obtained by repeated freeze-thawing of human plateletes, and the MSCs separated by density gradient centrifugation from 6 donors were expanded in medium supplemented with 10% fetal bovine serum (FCS) or HPL at different concentrations. The optimal concentration of HPL for cells culture was determined according to the cell proliferation kinetics. The cultured MSCs were characterized for their proliferation, cell phenotype, and cell cycle distribution.

RESULTSThe HPL-supplemented medium contained 4 essential growth factors for the growth of MSCs, namely platelet-derived growth factors (0.53∓0.06 ng/ml), basic fibroblast growth factor (37.5∓4.31 pg/ml), insulin-like growth factor-1 (0.15∓0.06 mg/ml) and transforming growth factor (5150∓463 pg/ml). Cultured in the presence of HPL at the optimal concentration of 7.5%, the MSCs displayed a spindle-shaped fibroblast-like morphology without obvious changes in the proliferation activity till passage 8 (P>0.05), similar to those of cells in FCS-supplemented culture medium. Flow cytometry and cell cycle analysis revealed no differences in the phenotypes or cell cycle distribution between the cells cultured in the presence of 7.5% HPL and 10% FCS.

CONCLUSIONThe culture medium supplemented by 7.5% HPL can promote the expansion of human MSCs and maintain the basic biological characteristics of the cells.

Blood Platelets ; cytology ; metabolism ; Cell Extracts ; pharmacology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Culture Media ; pharmacology ; Fibroblast Growth Factor 2 ; pharmacology ; Humans ; Mesenchymal Stromal Cells ; cytology ; drug effects ; Platelet-Derived Growth Factor ; pharmacology

OBJECTIVETo establish a predictive scoring system which may serve for the prediction of sustained response to conventional interferon-alpha (IFN-alpha) treatment on chronic hepatitis B.

METHODSA total of 474 IFN-alpha treated hepatitis B virus e antigen (HBeAg)-positive patients were enrolled in the present study. The patients' baseline characteristics, such as age, gender, aminotransferases, activity grading (G) of intrahepatic inflammation, score (S) of liver fibrosis, hepatitis B virus (HBV) DNA and genotype were evaluated; therapy duration and response of each patient at the 24th wk after cessation of IFN-alpha treatment were also recorded. A predictive scoring system for a sustained complete response (CR) to IFN-alpha therapy was established based on genetic algorithm. About 10% of the patients were randomly drawn out as the test set. Responses to IFN-alpha therapy were divided into CR, partial response (PR) and non-response (NR). The mixed set of PR and NR was recorded as PR + NR.

RESULTSFor the scoring system, the sensitivity and specificity were 78.8% and 80.6%, respectively.

CONCLUSIONThis SCR scoring system has satisfying prediction efficiency and is easily employed in clinical practice. With this scoring system, practitioners can propose individualized decisions that have an integrated foundation on both evidence-based medicine and personal characteristics.

Adolescent ; Adult ; Antiviral Agents ; therapeutic use ; Child ; Dose-Response Relationship, Drug ; Female ; Hepatitis B, Chronic ; drug therapy ; Humans ; Interferon-alpha ; therapeutic use ; Male ; Middle Aged ; Predictive Value of Tests ; Prognosis ; Sensitivity and Specificity ; Treatment Outcome ; Young Adult

BACKGROUNDTo construct the full-length complementary DNA of HCV genome from an HCV infected patient.

METHODSFour HCV gene fragments (1.6, 3.5, 2.4 kb and 2.6 kb) were amplified by RT-PCR from serum of a Chinese patient and fused and connected together to produce a 9.2 kb subgenomic fragment, which was further cloned into a cassette vector with fixed 5-prime and 3-prime termini of HCV to make the full-length cDNA. The cDNA heterogeneity was analyzed by comparing the sequences of 4 isolated HVR1 regions. The prokaryotic expressed Core, NS3 protease, NS3 helicase were detected for their specific reactivities with patient serum by Western blot analysis. And the protease activity of NS3 was evaluated in a cell-based NS3/4A-SEAP expression system.

RESULTSThe cDNA covered the near full-length of HCV genome from the patient's serum. The difference among HVR1 regions indicated no selection of HCV variants during RT-PCR and the quasi-species characteristic of the amplified cDNA. The prokaryotic expressed viral proteins could be identified by patient serum. In the NS3/4A-SEAP system, NS3 could cleave the 4A-4B site between NS4A and SEAP proteins and resulted in the secretion of SEAP in culture media.

CONCLUSIONThese results suggest that the cloned HCV cDNA encodes a complete and functional open reading frame and will be useful for further construction of infectious cDNA clone.

Base Sequence ; Cloning, Molecular ; DNA, Complementary ; genetics ; Genome, Viral ; Hepacivirus ; genetics ; Hepatitis C ; virology ; Humans ; Molecular Sequence Data ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo determine the distribution of HCV genotypes among volunteer blood donors in Guangzhou.

METHODSSix-nine HCV RNA-positive samples were collected from volunteer blood donors in Guangzhou. NS5B fragments of HCV were amplified followed by DNA sequencing and phylogenetic analysis.

RESULTSHCV genotypes were determined for 67 samples. Among them, the subtypes 1b, 2a, 3a, 3b, 6a and 6n were detected at the frequencies of 37.31%, 4.48%, 7.46%, 4.48%, 44.78% and 1.49%, respectively.

CONCLUSIONHCV 1b and 6a are the most predominant two subtypes among volunteer blood donors in Guangzhou.

Blood Donors ; China ; Genotype ; Hepacivirus ; classification ; genetics ; isolation & purification ; Humans ; Phylogeny ; RNA, Viral ; genetics ; Sequence Analysis, DNA