Objective To screen potential serum HCC associated proteins with low molecular weight and low abundance for better understanding the pathological mechanism of HCC and discovering new biomarkers.Methods All serum samples were collected from 81 HBV-related HCC patients,43 chronic hepatitis B patients and 36 cirrhosis patients.Serum protein fingerprint profiles were first generated by selected WCX2 protein chip integrating with SELDI-TOF-MS,and then normalized and aligned by Ciphergen SELDI Software 3.1.1 with Biomarker Wizard.Comparative analysis of the intensity of corresponding protein fingerprint peaks in normalized protein spectra was performed.Some protein peaks with significant difference among HCC,cirrhosis and chronic hepatitis B groups were found.The reproducibility of the SELDI system was assessed before serum protein fingerprint profiles analysis.Results The intra-and inter-assay CV for intensity and m/z in this SELDI system were 17.46% and 0.024%,and 17.74% and 0.024% respectively.Total 128 protein fingerprint peaks between 2 000 to 30 000 Da were identified under the condition of signal to noise＞5 and minimum threshold for cluster＞20%.Eighty-seven proteins were found to significantly expressed between HCC and cirrhosis groups(P＜0.05).Of the above differential proteins,forty-five proteins had changes greater than two fold,including 15 up-regulated proteins and 30 downregulated proteins in HCC sera.Between HCC and chronic hepatitis B groups,nine of fifty-two differential proteins(P＜0.05) had intensities of more than two folds,including 2 up-regulated proteins and 7 downregulated proteins in HCC sera.Between cirrhosis and chronic hepatitis B groups,twenty-eight of seventynine significantly differential proteins(P＜0.05) changed greater than two folds in intensity,including 17 up-regulated proteins in cirrhosis seru and 11 down-regulated proteins in chronic hepatitis B sera.Analysis of above leading differential proteins among three diseases using subtraction difference mode,the 5 common down-regulated proteins 2 870,3 941,2 688,3 165 and 5 483 m/z in HCC sera and 2 common up-regulated proteins 3 588 and 2 017 m/z in cirrhosis and HCC sera were screened.But no statistic difference in the level of protein 2 017m/z was found between HCC group and normal group inour previous study.Conclusion Because the interference of unspecific proteins from hepatitis B and cirrhosis could be eliminated partly in HCC sera through subtraction difference analysis,these 6 common differential proteins (2 870,3 941,2 688,3 165,5 483,3 588 m/z)have obvious advantages of increased specificity for evaluating the pathological state of HCC and might become promising candidate biomarkers in the diagnosis of HCC.

To study the effect of interleukin-15 (IL-15) on the proliferation, differentiation and apoptosis of MDS CD34(+) cells, CD34(+) cells of high enrichment were separated by MACS system, and cultured in liquid media with different concentration of IL-15 in treated group and without IL-15 in the control group. Apoptosis of hematopoietic precursors was assayed by propidium iodine staining and cell by FCM, and the other MDS CD34(+) cells were stained by cytochemical staining after culture. The results showed that after culture with IL-15 the proliferation and differentiation of MDS CD34(+) cells were obviously promoted. It was found the every lineage of mature cells developed, the expressions of cell surface antigens CD71, CD33 and CD19 all increased in the MDS CD34(+) cell treated with IL-15. It is suggested that IL-15 stimulates the proliferation and differentiation of MDS CD34(+) cells, and partly shows anti-apoptosis effects which may be applicable to the therapy MDS.
Antigens, CD ; immunology ; Antigens, CD19 ; immunology ; Antigens, CD34 ; immunology ; Antigens, Differentiation, Myelomonocytic ; immunology ; Apoptosis ; drug effects ; Bone Marrow Cells ; drug effects ; immunology ; pathology ; Cell Cycle ; drug effects ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Flow Cytometry ; Humans ; Interleukin-15 ; pharmacology ; Microscopy, Fluorescence ; Myelodysplastic Syndromes ; blood ; immunology ; pathology ; Receptors, Transferrin ; immunology ; Sialic Acid Binding Ig-like Lectin 3

Objective: To screen for the differentially expressed genes in laryngeal squamous cell carcinoma and normal laryngeal tissue using cDNA microarray. Methods: The PCR products of 4 096 genes were spotted on a chemical material coated glass plates in array. The DNAs were then fixed on the glass plate by a serial of treatments. The total RNAs were isolated from the tissues, and then were purified to mRNAs by Oligotex. Both the mRNAs from the laryngeal squamous cell carcinoma and normal tissue were reversely transcribed to cDNAs with the incorporations of fluorescent dUTP, for preparing the hybridization probes. The mixed probes were then hybridized to the cDNA microarray. After high stringent washing, the cDNA microarray was scanned for the fluorescent signals and showed the differences between 2 tissues. Results: Among the 4 096 target genes, there were 36(0.88%) genes whose expression levels differed between the carcinoma and normal tissues in all 4 cases. Bioinformatical analysis of those genes had been performed. Conclusion: DNA microarray technology is an effective technique in screening for differentially expressed genes between 2 different kinds of tissue. Further analysis of the obtained genes will help to understand the molecular mechanism of malignant carcinoma. [

Objective To observe the effect of low-frequency repetitive transcranial magnetic stimulation in improving the memory function and quality of life of patients with traumatic brain injury. Methods Sixty patients with brain injury were randomly divided into the treatmem group and control group. The patients in the control group received conventional treatments (including medications, acupuncture and exercise training), and those in the treatment group were given additional low-frequency repetitive transcranial magnetic stimulation treatment. Scores of the Rivermead Behavioral Memory Test (RBMT) and Functional independence Measure (FIM) were recorded before and after the treatments and compared between the two groups. Results The patients in both of the groups showed significant improvements in RBMT and FIM scores after the treatments (P<0.05). The improvements in RBMT and FIM scores after the treatments differed significantly between the two groups (P<0.05). Conclusion Low-frequency repetitive transcranial magnetic stimulation can significantly improve the cognitive function and functional independence of patients with traumatic brain injury.

Objective To introduce an improved method for collecting the blood of rat by cardiac puncture .Methods The study se-lected 90 Wistar rats,half male and half female ,and then the rats were anesthetized by intraperitoneally injecting 10％chloral hydrate solution according to 0.3 mL/kg body weight.Then using three-line positioning method to determine the puncture site ,the blood collection needle and blood collection tube were used to collecting the blood ,Finally,the blood volume of each rat was recorded .Results The success rate was cal-culated according to the single blood collection of 5 mL or more,and our 90 rats were collected 90 times,the success rate was about 82.2％. After centrifugation,the total serum was about 260 mL,the average serum separation of each rat was about 2.8 mL.Conclusion The im-proved positioning method of heart blood collection is intuitive and easy to learn ,and the total volume and quality of blood is high ,besides ,the new method has a small surgical trauma and a high efficiency ,which is worth to extend .

OBJECTIVE: To obtain regression formula for estimation of stature in Han population in Sichuan Province from length of the upper extremity long bones by digital radiography. METHODS: The statures of 365 healthy adults and digital radiographs of their right upper extremity long bones were measured. All statistical dispositions were done in SPSS including description and regression analysis. RESULTS: Twenty-seven simple regression and 3 multiple regression formulae with statistic significance in estimation of the stature from the sum of the length of the upper extremity long bones were established. Regression coefficient was higher in ulna than in radius, in male than in female, and was more accurate in multiple regression formulae than in simple regression formulae. CONCLUSION Digital radiograph measurement of the length of the upper extremity long bones seems to be a simple and practical method to estimate human stature in forensic practice.
Adult ; Aged ; Anthropometry ; Asian People ; Body Height ; Female ; Forensic Anthropology/methods* ; Humans ; Humerus/diagnostic imaging* ; Male ; Middle Aged ; Radiography ; Radius/diagnostic imaging* ; Regression Analysis ; Sex Characteristics ; Ulna/diagnostic imaging* ; Young Adult

Objective:To evaluate the influence of medical insurance payment standard reform on clinical drug use in designated medical institution.Methods:The medical expenses and drug consumption data of provincial level and Fuzhou medical insurance management center 3.31 million is included from April to September of 2016 and the same period of last year were statistically analyzed.The annual drug cost,drug use and the payment standard of med-ical insurance analysis were performed according to the annual statistics.Results: Before and after the reform, the proportion of the payment of standard drugs increased from 22.50%to 64.95%.The top 10 outpatient drugs in pub-lic hospitals and non-public hospitals were changed only in serial number,and after the reform,the cost of outpatient service fee was 73.21%and 39.30%respectively.Meanwhile,the variety and amount of drugs changed greatly of top 10 drugs public hospitals.The increase in drug cost of drugs with joint purchasing price difference was higher,and non-public hospitals were relatively stable.Conclusions: As per the findings of this study, Medicare insurance payment standard reform has significantly affected the choice of drug brands and drug use structure.To focus on the medical insurance payment standard of concentrated purchase of drugs combined with the amount of joint procurement of medicines is beneficial to reduce drug prices.The medical insurance payment standard reform combined with com-prehensive reform of medical and health system measures are much more conducive to play a guiding role.

OBJECTIVETo detect the RNA of severe acute respiratory syndrome virus (SARS-CoV) by using reverse transcription polymerase chain reaction (RT-PCR) targeted for a two loci and a modified nested real-time RT-PCR as to improving the reliability and sensitivity of tests.

METHODSA nested RT-PCR was used for detecting one fragment of SARS-CoV RNA in oropharyngeal swabs from 3 SARS probable patients, 4 SARS suspect patients and other 27 patients with fever in Hangzhou, and the nested RT-PCR product from one SARS probable patient was sequenced. Meanwhile in these 3 SARS probable patients, other three RT-PCR methods, including a hemi-nested RT-PCR targeted for another fragment of SARS-CoV RNA, a real-time RT-PCR and a modified nested real-time RT-PCR, were employed to detect SARS-CoV RNA.

RESULTSTwo positives were found in the 3 SARS probable patients, and none positive in 4 SARS suspect patients and other 27 patients with fever, using the nested RT-PCR. The sequence of the nested RT-PCR product from one SARS probable patient was identified with the counterpart of SARS-CoV genomes published in public database. The results of the hemi-nested RT-PCR, the real-time RT-PCR and the modified nested real-time RT-PCR in the 3 SARS patients were consistent with the one of the nested RT-PCR. During detecting specimen with low copies of RNA, a weak positive signal was produced after about 35 cycles in the real-time RT-PCR, but a strong positive signal was found only after 10 cycles in the modified nested real-time RT-PCR.

CONCLUSIONIt might improve the reliability of test by employing RT-PCR targeted for two or more fragments in SARS-CoV genome. The modified nested real-time RT-PCR might have higher sensitivity than the routine real-time RT-PCR.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Base Sequence ; Humans ; Middle Aged ; RNA, Viral ; genetics ; metabolism ; Reproducibility of Results ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; SARS Virus ; genetics ; Sensitivity and Specificity ; Severe Acute Respiratory Syndrome ; diagnosis ; virology ; Young Adult

OBJECTIVEDendritic cells an hyperinsulinemia are both implicated in the pathogenesis of atherosclerosis. The aim of this study is to explore the effect of high concentration of insulin on the maturation of monocyte-derived dendritic cells (MoDCs) and related signal transduction pathways.

METHODSHuman monocytes were purified (over 98%) using Anti-CD14 micro-beads and cultured for 5 days with DC Cellgro medium containing rhGM-CSF (100 microg/L) and rhIL-4 (20 microg/L). Immature DC were then incubated with insulin of various concentrations (0, 1, 10, 100 nmol/L) for 24 hours in the presence or absence of LY294002 (PI3K inhibitor) or PD98059 (MAPK inhibitor). Immunophenotypic expression of CD86 and CD83 were detected using flow cytometry. Endocytosis function of the MoDCs was evaluated using FITC-Dextran and MoDCs secretion IL-12, IFN-gamma and TNF-alpha were measured by ELISA.

RESULTSInsulin induced significantly higher CD83 and CD86 expressions on MoDCs in a dose-dependent manner. The endocytosis function of MoDCs were significantly inhibited and cytokine secretions of IL-12, IFN-gamma and TNF-alpha significantly increased by 10 nmol/L and 100 nmol/L insulin. These effects could be blocked by the LY294002 and PD98059.

CONCLUSIONHyperinsulinemia contributed to atherosclerosis via stimulating immune maturation of MoDCs via both PI3K and MAPK pathways.

Cell Differentiation ; drug effects ; immunology ; Cells, Cultured ; Cytokines ; metabolism ; Dendritic Cells ; drug effects ; immunology ; metabolism ; Humans ; Insulin ; administration & dosage ; pharmacology ; Monocytes ; cytology ; Phagocytosis ; drug effects ; Signal Transduction

OBJECTIVETo investigate the protective effect of L-carnitine (LC) combined with sildenafil on the reproductive endocrine function of male rats with diabetes mellitus (DM).

METHODSA total of 40 male SD rats were randomly divided into five groups, group A taken as normal controls, and groups B, C, D and E made into DM models by injection of streptozotocin at 65 mg/kg. Then the rats in groups A and B were treated with normal saline, C with sildenafil at 5 mg per kg per d, D with LC at 300 mg per kg per d, and E with sildenafil at 5 mg per kg per d plus LC at 300 mg per kg per d, all via gastric gavage for 6 weeks, followed by determination of the levels of testosterone (T), follicle-stimulating hormone (FSH) and luteinizing hormone (LH) in the serum of the rats.

RESULTSAfter 6 weeks of treatment, the T, FSH and LH levels were (25.25 +/- 2.67) nmol/L, (5.78 +/- 0.61) IU/L and (625.21 +/- 43.45) ng/L in group A, (9.63 +/- 1.71) nmol/L, (1.98 +/- 0.42) IU/L and (479.89 +/- 27.62) ng/L in group B, (18.98 +/- 3.07) nmol/L, (5.08 +/- 0.33) IU/L and (586.57 +/- 31.72) ng/L in group C, (16.18 +/- 2.65) nmol/L, (4.63 +/- 0.30) IU/L and (540.78 +/- 25.52) ng/L in group D, and (23.65 +/- 2.66) nmol/L, (5.59 +/- 0.48) IU/L and (621.53 +/- 36. 40) ng/L in group E. The three parameters were significantly lower in B than in the other four groups (P < 0.01), and so were they in C and D than in A and E (P < 0.05), but showed no significant differences either between C and D (P > 0. 05) or between A and E (P > 0.05).

CONCLUSIONSix-week medication of either sildenafil or LC alone could increase the levels of T, FSH and LH in the serum of DM rats, but the combination of the two had an even more obvious increasing effect, which indicates a still better protective effect on the reproductive endocrine function of diabetic male rats.

Animals ; Carnitine ; adverse effects ; therapeutic use ; Diabetes Mellitus, Experimental ; drug therapy ; metabolism ; Drug Therapy, Combination ; Follicle Stimulating Hormone ; blood ; Luteinizing Hormone ; blood ; Male ; Piperazines ; administration & dosage ; therapeutic use ; Purines ; administration & dosage ; therapeutic use ; Rats ; Rats, Sprague-Dawley ; Sildenafil Citrate ; Sulfones ; administration & dosage ; therapeutic use ; Testosterone ; blood