Objective: The current study was designed to distinguish Phase G0/G1， S， G2/M cell groups in the whole cell cycle by DNA flow cytometry. Method: The authors developed a multiparameter flow cytometry of cyclin E＋ A/DNA for MOLT-4. Result: The data showed that the method could distinguish Phase G0, early G1, late G1, S, G2,and M cells. The method could be used in molecular cell biology, especially in cell kinetic study.

OBJECTIVETo investigate the therapeutic efficacy of interferon (IFN) therapy and risk of long-term administration for chronic hepatitis C (CHC) patients who cannot tolerate the standard treatment.

METHODSForty-six CHC patients who had proven intolerant to standard treatments were treated with low-dose IFN (non-pegylated IFN: 60 to 300MIU QOD, or pegylated IFN: 50 to 90 mug/w) plus ribavirin (RBV; 0.6g to 0.9 g/d) for 72 weeks.

RESULTSForty-three (93.5%) of the patients were able to tolerate the long-term treatment with low-dose IFN plus RBV. Only three patients experienced severe side effects (low white blood cell and platelet counts) that required treatment withdrawal. The virology response rates over treatment time were: rapid virologic response (RVR): 10.9%; early virus response (EVR): 30.4%; 24 week virologic response: 45.7%; and, 48 week virologic response: 47.8%. B-sonographic imaging revealed that three patients experienced improved liver morphology through the treatment course. The patients who achieved RVR, EVR, or 24 weeks virologic response also attained higher 48 week virologic response. The 24 week virologic response had the strongest predictive value of good prognosis.

CONCLUSIONSOur study demonstrated that long-term treatment with low-dose interferon plus ribavirin is effective for patients who are otherwise intolerant to standard treatment. In these patients, low-dose IFN plus RBV can obtain a high virologic response rate at 48 week. Furthermore, the 24 week virologic response is sufficiently predictive of treatment success. As with any treatment regimen, it is important for healthcare workers to monitor the disease status and potential side effects throughout the course of therapy.

Adult ; Antiviral Agents ; administration & dosage ; therapeutic use ; Female ; Hepacivirus ; Hepatitis C, Chronic ; drug therapy ; virology ; Humans ; Interferons ; administration & dosage ; therapeutic use ; Male ; Middle Aged ; Treatment Outcome

Animals ; History, 20th Century ; Hong Kong ; epidemiology ; Humans ; Influenza A Virus, H3N2 Subtype ; classification ; genetics ; isolation & purification ; Influenza, Human ; epidemiology ; history ; virology ; Molecular Sequence Data ; Phylogeny

AIM To study the phenylethanoid glycosides from Verbenae Herba.METHODS The 80%ethanol extract from Verbenae Herba was isolated and purified by silica gel,Sephadex LH-20,TLC and semi-preparative HPLC,then the structures of obtained compounds were identified by physicochemical properties and spectral data.RESULTS Nine compounds were isolated and identified as verbofficoside A(1),cistanoside D(2),epimeredinoside A(3),verbascoside(4),isoverbascoside(5),cistanoside C(6),cistanoside F(7),decaffeoylacteoside(8),jionoside C(9).CONCLUSION Compound 1 is a new compound.Compounds 3 and 6-9 are isolated from this plant for the first time.

OBJECTIVETo summarize the experience in the management of slow transit constipation complicated with adult megacolon.

METHODSThe clinical data of 32 above patients admitted between October 2007 and June 2011 were retrospectively studied.

RESULTSThirty-two patients were diagnosed as slow transit constipation according to the Roman III criteria. There were 15 males and 17 females aging from 18 to 56 years old. Sitz marker study showed prolonged colon transit time. Barium enema and defecography suggested bowel stricture locating in the transverse colon (n=3), descending colon (n=4), rectum (n=20), and concurrent transverse colon or descending colon and rectum (n=5). Anal manometry showed that anorectal inhibitory reflex was absent in 23 patients, while the other 9 patients were normal. Procedures performed included segmental colectomy and side-to-side anastomosis (n=1), subtotal colectomy and modified Duhamel anastomosis (n=16), total colectomy and ileal J-pouch Duhamel anastomosis (n=9). There were no postoperative complications. During the follow-up ranging from 3 to 47 months, defacatory function was excellent in 18, good in 9, and moderate in 5 patients.

CONCLUSIONSAdult megacolon should be considered differential diagnosis of slow transit constipation. Detailed history taking and thorough evaluation of testing is the key to obviate misdiagnosis. Extent of resection should include the diseased dilated colon and slow transit colon.

Adolescent ; Adult ; Aged ; Anastomosis, Surgical ; Constipation ; etiology ; surgery ; Digestive System Surgical Procedures ; Female ; Gastrointestinal Transit ; Humans ; Intestinal Obstruction ; Male ; Megacolon ; complications ; Postoperative Complications ; Retrospective Studies

OBJECTIVETo assess the efficacy and safety of intravascular ultrasound (IVUS)-guided interventional therapy for borderline lesions in patients with acute coronary syndrome (ASC).

METHODSThirty-one ASC patients with borderline lesions (coronary artery stenosis between 40%-70% confirmed by coronary arteriography [CAG]) and a minimal lumen area (MLA) of the infarction related artery(IRA) < or =4.0 mm(2) shown by IVUS underwent percutaneous coronary intervention (PCI). Another 31 PCI cases without IVUS were also enrolled as the control group. The minimal luminal diameter, cross section luminal area, total cross section, plaque area and area stenosis rate were measured before and after stent deployment at a conventional or higher pressure in the IVUS group. All the patients were followed up for 10-12 months and clinically evaluated 1, 3, 6 month and 12 months after the procedure to collect the data of angina recurrence, myocardial infarction and revascularization.

RESULTSAll the 31 cases were successfully stented with satisfied CAG results (with residual stenosis <0, TIMI flow grade III) and without dissection or any related complications. Among the 32 stents, 28 showed insufficient adherence or underexpansion (stent malapposition) to require 18-20 atm dilation or another high pressure balloon to attain the adequate IVUS results. CAG and IVUS were repeated in 22 patients (70.97%) of the IVUS group during the 10 to 12 months of follow up. No stent restenosis occurred with the in-stent diameter late loss >50%, nor was in-stent thrombus found by IVUS. Endomembrane proliferation was found but without any significant difference. Minimal stent lumen area were not significantly different from the immediate results after PCI (10.12-/+1.15 mm(2) vs 8.98-/+2.12 mm(2), P>0.05). The 31 patients in the control group were successfully stented with satisactory CAG results, but 3 suffered angina at 3-6 months who showed stent restenosis and insufficient stent adherence.

CONCLUSIONIVUS can more effectively guide the interventional therapy for ACS borderline lesions and assess the immediate efficacy of therapy than CAG. Post-dilation with higher pressure (16-20 atm) guided by IVUS can further improve the procedural results. IVUS-guided PCI for ACS borderline lesions ensures high immediate and long-term success rate.

Acute Coronary Syndrome ; therapy ; Adult ; Aged ; Angioplasty, Balloon, Coronary ; methods ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Stents ; Ultrasonography, Interventional

BACKGROUNDPatients with myelodysplastic syndrome (MDS) display a very diverse pattern. In this study, we investigated prognostic factors and survival rate in adult patients with MDS refractory anaemia (MDS-RA) diagnosed according to French-American-British classification and evaluated the International Prognostic Scoring System (IPSS) for Chinese patients.

METHODSA multi-center study on diagnosis of MDS-RA was conducted to characterize the clinical features of Chinese MDS patients. The morphological criteria for the diagnosis of MDS-RA were first standardized. Clinical data of 307 MDS-RA patients collected from Shanghai, Suzhou and Beijing from 1995 to 2006 were analyzed using Kaplan-Meier curve, log rank and Cox regression model.

RESULTSThe median age of 307 MDS-RA cases was 52 years. The frequency of 2 or 3 lineage cytopenias was 85.6%. Abnormal karyotype occurred in 35.7% of 235 patients. There were 165 cases (70.2%) in the good IPSS cytogenetic subgroup, 44 cases (18.7%) intermediate and 26 cases (11.1%) poor. IPSS showed 20 (8.5%) categorized as low risk, 195 cases (83.0%) as intermediate-I risk and 20 cases (8.5%) as intermediate-II risk. The 1-, 2-, 3-, 4- and 5-year survival rates were 90.8%, 85.7%, 82.9%, 74.9% and 71.2% respectively. Fifteen cases (4.9%) transformed to acute myeloid leukaemia (median time 15.9 months, range 3 - 102 months). Lower white blood cell count (< 1.5 x 10(9)/L), platelet count (< 30 x 10(9)/L) and cytogenetic abnormalities were independent prognostic factors by multivariate analysis, but age (= 65 years), IPSS cytogenetic subgroup and IPSS risk subgroup were not independent prognostic factors associated with survival time.

CONCLUSIONSChinese patients were younger, and had lower incidence of cytogenetic abnormalities, more severe cytopenias but a more favourable prognosis than Western patients. The major prognostic factors were lower white blood cell count, lower platelet count and fewer abnormal karyotypes. The international prognostic scoring system risk group was not an independent prognostic factor for Chinese myelodysplastic syndrome patients with refractory anaemia patients.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Anemia, Refractory ; etiology ; mortality ; Asian Continental Ancestry Group ; Child ; China ; Female ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; complications ; mortality ; Prognosis

OBJECTIVETo construct the cDNA subclones spanning the entire genome of dengue 2 virus NGC strain for further construction of full-length infectious viral cDNA clone.

METHODSTwo pairs of primers were designed according to the restriction endonuclease sites in the viral genome of dengue 2 virus NGC strain. After viral RNA extraction from the brain of infected new-born mice, two parts of full-length viral cDNA were amplified by long RT-PCR and cloned into the vector pCR-XL-TOPO. The partial sequence of the recombinant plasmid was determined.

RESULTS AND CONCLUSIONSequence analysis and digestion with restriction enzymes demonstrated that the two cDNA subclones were specific for dengue 2 virus NGC strain, suggesting the successful construction of the two cDNA subclones of dengue 2 virus NGC strain.

Animals ; Animals, Newborn ; Brain ; virology ; Cloning, Molecular ; DNA, Complementary ; biosynthesis ; genetics ; DNA, Viral ; biosynthesis ; genetics ; Dengue ; virology ; Dengue Virus ; classification ; genetics ; isolation & purification ; Genome, Viral ; Mice ; RNA, Viral ; genetics ; isolation & purification ; Recombination, Genetic ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo determine the sequence of S2 gene of SARS-associated coronavirus (SARS-CoV) GD322 and analyze the phyletic evolution of S2 gene.

METHODS2 gene fragment was amplified from SARS-CoV GD322 genome with RT-PCR and ligated to pGEM-T vector for sequence analysis after transformation of the plasmid into E. coli DH5a. The variability of S2 genes and S2 proteins from 12 strains isolated in the early, intermediate and advanced stages of the SARS outbreak were analyzed and the phylogenetic tree was constructed with Lasergene, Clustal X, DNAman and Treeview. T cell antigen epitopes of S2 protein were predicted on the basis of Internet database.

RESULTWith the epidemic spread of SARS-CoV, the S2 genes of the virus tended to become stable. Homology of S2 genes of SARS-CoV isolated in advanced stage of the outbreak reached 99.9%. Prediction of T cell antigen epitope showed that mutation at the 57th amino acid effected T cell antigen epitope.

CONCLUSIONS2 gene of GD322 SARS-CoV is relatively stable during the epidemic spread of the virus, and mutation at the 57th amino acids of S2 protein may affect the T cell antigen epitope.

Escherichia coli ; genetics ; Genetic Variation ; Humans ; Phylogeny ; Point Mutation ; SARS Virus ; genetics ; isolation & purification ; Sequence Analysis, DNA ; Severe Acute Respiratory Syndrome ; virology ; Viral Envelope Proteins ; genetics

This study was purposed to investigate the effect of multiple myeloma patients' sera on hepcidin mRNA expression of Hep-3b hepatoma cell line and effect of human interleukin-6 (IL-6) antibody or recombinant human erythropoietin (rhEPO) on hepcidin mRNA expression. The clinical information and serum of multiple myeloma patients were collected. Their sera of a final concentration of 10% were added into Hep-3b cell medium. The mRNA from Hep-3b cells was extracted, and hepcidin mRNA expression was detected by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). A final concentration of 10 ng/ml human IL-6 antibody and 2 U/ml rhEPO were added into the medium respectively. The results showed that the sera of untreated multiple myeloma patients elevated hepcidin mRNA expression of Hep-3b cells, compared with healthy controls and iron deficiency anemia patients. This effect was fully neutralized by human IL-6 antibody or rhEPO. The hemoglobin (Hb) level was stable during the follow up of regularly treated multiple myeloma patients and the effect of MM patient serum on Hep-3b cell hepcidin mRNA expression was reduced. It is concluded that the hepcidin mRNA expression of Hep-3b cell can be increased by untreated multiple myeloma patient serum. This promotive effect can be antagonised by IL-6, which suggests that IL-6 may be possible to elevate expression level of hepcidin in Hep-3b cells and results in anemia of chronic disease (ACD). The above mentioned promotive effects also can be suppressed by rhEPO, which indicates that the rhEPO may possess curative effect for ACD disease. During short-term follow-up of treated patients with multiple myeloma the Hb level is stable, the influence of patients serum on hepcidin mRNA of Hep-3b cells decreases, which shows the stabilization of disease and amelioration of ACD patient status.
Adult ; Aged ; Antibodies, Monoclonal ; pharmacology ; Antimicrobial Cationic Peptides ; genetics ; Cell Line, Tumor ; Erythropoietin ; blood ; pharmacology ; Female ; Hepcidins ; Humans ; Interleukin-6 ; immunology ; Male ; Middle Aged ; Multiple Myeloma ; genetics ; metabolism ; RNA, Messenger ; genetics