Objective To find a proper way for accurate results on completing blood counts.Methods Based on the results from automatic hematology analyzer XE-2100,set up the criteria for blood cell microscopic examination.1368 blood specimen were detected and the results were analyzed according to the criteria.Statistics on the data were made to evaluate the accordance between warnings of analyzer and manual examination,likewise the reliability of the criteria.Results Comparing with microscopic examination,analyzer warning on low PLT has good accordance,Kappa value was 0.95,U value was 35.19,P

Objective To develop an effective way to evaluate the accurate platelet count in a patient with anticoagulants-induced pseudothrombocytopenia (PTCP).Methods It was studied that various anticoagulants effect on the platelets count for an infrequent patient with anticoagulants-dependent PTCP. When vitamin B6,aminophylline,gentamicin and amikacin were separately added to four anticoagulated blood samples from anticoagulants-dependent patient within 15 min after blood withdrawal,platelets count and morphological changes of blood cells after 4 hours of incubation at room temperature were investigated. The best anti-aggregating agent and its optimal concentration among them were explored.Results The four anticoagulants all could not inhibit the aggregation of the patient's platelets.Only amikaein among the above anti-aggregating agents can prevent and dissociate the aggregation of platelets without apparent morphological changes of blood cells and the platelet counts was stable within 4 hours after blood drawn when amikacin was added either before or after blood sampling.With increasing the concentration of amikaein,the platelet counts increase and then tend to be stable.The optimal concentration of amikacin is 5 mg/ml blood.Conclusions The supplementation of amikaein either before or after blood sampling is a useful method for the diagnosis anticoagulants-dependent PTCP and for the eva/uation of platelet counts in infrequent patients with anticoagulants-dependent PTCP.

In order to demonstrate the taxonomic position of paramyxovirus Tianjin strain and explore its mechanism of pathogenesis. Bioinformatics methods were used to analyze the deduced amino acid sequences of NP, P, M, and L protein of Tianjin strain. Phylogenetic analysis based on NP, P, M, and L protein sequences demonstrated that Tianjin strain belonged to the genus Respirovirus, in the subfamily Paramyxovirinae and most likely a new genotype of Sendai virus. Sequence similarities comparisons indicated that Tianjin strain P protein was poorly conserved, sharing only 78.7%~91.9% amino acid identity with 6 known Sendai viruses, while L protein was the most conserved, having 96.0%~98.0% amino acid identity with other Sendai viruses. Multiple-sequence alignments of Tianjin strain NP, P, M, and L protein with those of 6 known Sendai viruses showed that Tianjin strain possessed a lot of unique amino acid substitutions in protein sequences, 15 in NP, 29 in P, 6 in M, and 29 in L. The presence of these unique amino acid substitutions suggests that Tianjin strain maybe has a significant difference in host or pathological characteristics from the known Sendai viruses.

To culture bone marrow mesenchymal stem cells (BMSCs) of rat in vitro and observe HSV-1 infection on BMSCs, BMSCs were separated from the bone marrow and identified by alizarin red staining and detection of ALP. The morphology of HSV-1 infected BMSCs and the CPE were observed. The total DNA was extracted from HSV-1 infected BMSCs and the desired specific gene fragment of 477bp of HSV-1 was amplified by PCR. Results showed that after BMSCs were induced by mineral-fluid for 14 days, the ALP level was increased and the nodule calcification was formed. The induced BMSCs were manifested to have the characteristics of osteoblasts. CPE couldn't be found in HSV-1 latently infected BMSCs but the 477bp gene fragment was still detectable. HSV-1 could establish latent infection in BMSCs after 7 passages. This study indicated that rat BMSCs could be induced to differentiate into osteoblasts in vitro, therefore they can be used as the seed cells for the tissue engineering. HSV-1 can infect rat BMSCs and develop the latent infection in vitro.
Alkaline Phosphatase ; analysis ; Animals ; Bone Marrow Cells ; virology ; Cell Differentiation ; Female ; Herpesvirus 1, Human ; physiology ; Male ; Mesenchymal Stromal Cells ; cytology ; virology ; Polymerase Chain Reaction ; Rats ; Rats, Wistar ; Tissue Engineering ; Virus Latency

OBJECTIVETo compare the prognosis of small hepatocellular carcinoma patients with hepatitis B virus infection versus hepatitis C virus infection.

METHODS413 patients receiving curative resections at Tianjin Cancer Hospital for small HCC (< or = 3 cm) from January 1997 to December 2003 were divided into four groups: HCV only (n = 75), HBV only (n = 251), HBV and HCV (n = 33), and neither HBV nor HCV (NBNC, n = 54). The preoperative status and postoperative recurrence were recorded. Survival analysis were used to assess the impact of HBV/HCV status on HCC recurrence.

RESULTSPatients with HCV were associated with older age, lower mean preoperative platelet counts and albumin levels, higher mean prothrombin time, alanine aminotransferase and total bilirubin levels. Tumors in patients with HCV are multinodular and less differentiated, and were associated with a higher incidence of vascular invasion and cirrhosis. During the follow-up, the HCV group showed a higher incidence of intrahepatic recurrence and multiple recurrent lesions than the other patients.

CONCLUSIONSHCC patients with HCV infection tended to be older, and were characterized by more severe cirrhosis and higher incidence of tumor multinodular. The statistically significant determinants of reoccurrence in patients with small HCC were HCV infection, presence of vascular invasion and multiple tumors.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Carcinoma, Hepatocellular ; pathology ; surgery ; virology ; Female ; Follow-Up Studies ; Hepatectomy ; Hepatitis B ; complications ; epidemiology ; Hepatitis C ; complications ; epidemiology ; Humans ; Liver Cirrhosis ; pathology ; virology ; Liver Neoplasms ; pathology ; surgery ; virology ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; epidemiology ; Prognosis ; Retrospective Studies ; Risk Factors ; Survival Analysis ; Young Adult

Background The excitotoxicity to retinal neurons caused by abnormal elevation of glutamate in retina is a common pathology concomitant with major blind-causing eye diseases.However,an effective approach to protect retinal neurons from glutamate-induced excitotoxicity is still lack.Intraperitoneal administration of α-melanocyte stimulating hormone(α-MSH)has been shown to protect hippocampal neurons from glutamate-induced excitotoxicity.Objective This study was to investigate the protective effect of α-MSH on glutamate-induced excitotoxicity in a chicken embryonic retinal explant culture system.Methods The retinas were isolated from chick embryos at embryonic day 9(E9) and cultured as explants.The explants at 3,5 and 7 days in vitro and the retinas at corresponding embryonic day 12,14 and 16(E12,E14,E16)were collected.The morphology of explant cultures was examined by hematoxylin and eosin staining,and the expression of melanocortin receptors (MCRs)was analyzed by real-time PCR.In the experiment of glutamate-induced excitotoxicity,the retinal explants at 4 days in vitro were treated with glutamate for 48 hours,α-MSH was incubated with the explants 30 minutes before and during the glutamate treatment period.Then the apoptotic cells were detected by TUNEL staining and quantified.The glutamate alone treated-explants and those treated with culture media were included as controls.The expression of glial fibrillary acidic protein(GFAP) at 48 hours after treatment in all retinal explants was analyzed by real-time PCR.Results Hematoxylin and eosin staining showed that the retinal explants exhibited similar morphology to those observed in the retinas from chick embryos at the corresponding developmental stages.The real-time PCR analyses of chick retinas showed that MC1R mRNA level at E9,E12,E14 and E16 was significantly lower than that in post-hatch day 1 (all P=0.000) ;whereas the transcript level of MC5R was significantly increased from E9 to E12 and E14 (both P =0.000),and then gradually decreased from E14 to P1.The expression of these genes showed similar temporal patterns in the retinal explant cultures.TUNEL staining revealed that treatment of the retinal explant cultures with α-MSH substantially and significantly reduced number of apoptotic cells induced by glutamate (P =0.000),which was accompanied by significant suppression of glutamate-induced GFAP up-regulation (P =0.000).Conclusions Application of α-MSH dramatically ameliorated glutamate-induced cell death in retinal explant cultures.This protective effect may be due to α-MSH-mediated suppression of astrogliosis caused by abnormal elevation of glutamate.

Paramyxovirus Tianjin strain is the high-pathogenic virus to primate and might also cause human lower respiratory tract infection. To determine the genome structure, variation features and phylogenetic position, the complete nucleotide sequence of paramyxovirus Tianjin strain was analyzed. The homology comparison and phylogenetic analysis of the nucleotide and the deduced amino acid sequences among paramyxovirus Tianjin strain and the 28 strains in seven genera and the 7 unclassified viruses of Paramyxoviridae were performed. The results suggested that Tianjin strain is a member of the Respirovirus genus in the Paramyxovirinae, Paramyxoviridae and has the closest relationship to Sendai virus. Its genome length and composition are similar to the previously published Sendai virus except one extra glutamic acid residue increasing at the C terminus of Large protein due to the genomic RNA mutation at position A15240C. 440 unique nucleotide variations of Tianjin strain lead to 110 amino acid residue changes, making it differed from any other Sendai viruses. The phylogenetic analysis reveals paramyxovirus Tianjin strain doesn't belong to any of the three known evolution lineages of Sendai viruses and locates at a separate evolution branch. The obvious distinctions of genome nucleotide sequence, host tropism and pathogenicity suggest that paramyxovirus Tianjin strain might represent a novel genotype of Sendai virus.
Base Sequence ; Evolution, Molecular ; Genome, Viral ; Paramyxoviridae ; classification ; genetics ; Phylogeny ; Polymerase Chain Reaction ; RNA, Viral ; chemistry ; Sendai virus ; genetics

OBJECTIVETo observe the effects of combined treatment with sansanmycin and macrolides on Pseudomonas aeruginosa and formation of biofilm.

METHODSMicro-dilution method was used to determine the minimal inhibitory concentrations (MICs) of sansanmycin, gentamycin, carbenicillin, polymyxin B, roxithromycin, piperacillin, and tazobactam. PA1 and PA27853 biofilms were observed under optical microscope after staining and under SEM after treatment with sansanmycin at different dosages and combined treatment with sansanmycin and roxithromycin. Viable bacteria in PA1 and PA27853 biofilms were counted after treatment with sansanmycin at different dosages or combined treatment with sansanmycin and roxithromycin.

RESULTSThe MIC of sansanmycin was lower than that of gentamycin and polymyxin B, but was higher than that of carbenicillin. Roxithromycin enhanced the penetration of sansanmycin to PA1 and PA27853 strains through biofilms. PA1 and PA27853 biofilms were gradually cleared with the increased dosages of sansanmycin or with the combined sansanmycin and roxithromycin.

CONCLUSIONSub-MIC levels of roxithromycin and sansanmycin substantially inhibit the generation of biofilms and proliferation of bacteria. Therefore, combined antibiotics can be used in treatment of intractable bacterial infection.

Animals ; Anti-Bacterial Agents ; administration & dosage ; pharmacology ; Bacterial Adhesion ; drug effects ; Biofilms ; growth & development ; Cercopithecus aethiops ; Drug Therapy, Combination ; Macrolides ; administration & dosage ; pharmacology ; Microbial Sensitivity Tests ; Oligopeptides ; administration & dosage ; pharmacology ; Pseudomonas aeruginosa ; drug effects ; physiology ; ultrastructure ; Uridine ; administration & dosage ; analogs & derivatives ; pharmacology ; Vero Cells

The methods to determine the total phenols, total saponins, and marker constituents salidroside, chlorogenic acid and 3, 4-dihydroxy-phenylethyl-β-D-glucopyranoside in the samples of Sargentodoxae Caulis were established to provide the evidence for the improvement and revision of the quality standard of the crude material recorded in the Chinese Pharmacopoeia (2015 Edition). The content of total phenols was determined by ultraviolet spectrophotometry, using gallic acid as a reference substance. The content of total saponins was determined by ultraviolet spectrophotometry, using 3-O-[β-D-xylopyranosyl-(1-2)-O-β-D-glucuronopyranosyl]-28-O-[β-D-glucopyranosyl] asiatic acid as a reference substance. The contents of salidroside, chlorogenic acid and 3,4-dihydroxy-phenylethyl-β-D-glucopyranoside were detected by HPLC. The linear ranges were 1.01-7.04 mg x L(-1) for total phenols, 37.7-201 μg for total saponins, 0.025 8-1.55 μg for salidroside, 0.076 2-5.44 μg for chlorogenic acid, and 0.064 9-3.47 μg for 3,4-dihydroxy-phenylethyl-βP-D-glucopyranoside, respectively. Their average recoveries were 99.12%, 99.11% 105.5%, 99.08%, and 101.6%, respectively. The contents of total phenols and total saponins were 3. 04% -11. 9% and 0. 87% -3. 63%. The contents of salidroside, chlorogenic acid and 3,4-dihydroxy-phenylethyl-β-D-glucopyranoside fluctuated from 0.018% to 0. 572%, from 0.041% to 1.75% and from 0.035% to 1.32%. The established methods were reproducible, and they could be used for the quality control of Sargentodoxae Caulis. The present investigation suggested that total phenols, salidroside, and chlorogenic acid should be recorded in the quality standard of Sargentodoxae Caulis and their contents should not be less than 6.8% for total phenols, 0.040% for salidroside, and 0.21% for chlorogenic acid.
China ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; Magnoliopsida ; chemistry ; Phenol ; analysis ; Plant Stems ; chemistry ; Saponins ; analysis ; Triterpenes ; analysis

OBJECTIVETo clarify the dose-response relationship between asbestos dust exposure and lung cancer incidence in chrysotile asbestos miners by fixed cohort study and to investigate the incidence rates of lung cancer in exposure to different concentrations of asbestos dust.

METHODSA retrospective cohort study was conducted in 1932 asbestos miners who registered from January 1, 1981 to December 31, 1988, had worked for at least 1 year, and had no obvious cardiopulmonary diseases; the cohort study began in July 2009 and covered a time span of 29 years (1981 - 2009). The personal information, occupational history, disease history, and health data of these miners were recorded, and the monitoring data on dust concentrations in the mine over the years were collected. The dose-response relationship between asbestos dust concentration and lung cancer incidence was established by the method of life table; a regression equation was fitted to predict the excess incidence rates of lung cancer under the conditions of different working years and dust concentrations.

RESULTSA significant dose-response relationship was observed between cumulative exposure (Ce) and cumulative probability (Px) of lung cancer incidence, and the smokers hada higher Px than nonsmokers. When Ce was less than 2000 mg/m(3)·each year, Px reached 6.58/10000; when Ce was not less than 2000 mg/m(3)·and less than 3000 mg/m(3)·each year, Px reached 91.72/10000; when Ce was more than 5000 mg/m(3)·each year, Px was as high as 141.02/10000. The three models were fitted to obtain the optimal regression equation: Px = -0.0004Ce(2) + 0.0052Ce - 0.0011 (r(2) = 0.9387). In the workshop of asbestos mine in this study, the average dust concentration was 85 times higher than the limit in 2009, so the excess incidence rate of lung cancer was 112.598/10000 if the miners worked under this condition for 40 years, according to the equation.

CONCLUSIONThere is a significant dose-response relationship between cumulative asbestos exposure and lung cancer incidence in chrysotile asbestos miners. The risk for lung cancer rises as asbestos exposure increases.

Asbestos, Serpentine ; toxicity ; Dust ; Female ; Humans ; Lung Neoplasms ; etiology ; Male ; Mining ; Occupational Exposure ; Retrospective Studies