OBJECTIVETo study the relationship between TCM syndrome type and HBV-DNA in serum and peripheral blood mononuclear cells (PBMCs) in chronic hepatitis B (CHB) patients.

METHODSThe serum HBV markers,HBV-DNA levels in serum and PBMCs, were quantitatively detected in 220 CHB patients by PCR method, and TCM syndrome type of 205 patients were differentiated.

RESULTSArranged from low to high, the percentages of CHB patients with the serum HBV-DNA > or = 1.0 x l0(5) copy/mL (high viral loading) in the five syndrome types were as follows: damp-heat retention in middle-jiao syndrome (DHRS, 55.2%), blood stasis blocking collateral syndrome (BSBC), Gan-Shen yin deficiency syndrome (GSYS), Pi-Shen yang deficiency syndrome (PSDS) and Gan stagnation with Pi deficiency syndrome (GSPS, 82.5%), the difference was significant between DHRS and GSPS; those with HBV-DNA in PBMCs infection were: GSYS (27.3%), DHRS (34.3%), BSBC (53.1%) and GSPS (77.2%). The percentage in GSPS was the highest, which was significantly different to that in other syndromes.

CONCLUSIONAmount of serum HBV-DNA and PBMCs HBV-DNA infection has certain correlation with the TCM syndrome type of CHB. The highest percentage of patients with HBV-DNA > or = 1.0 x l0(5) copy/mL and PBMCs HBV-DNA infection presented in CHB patients of GSPS type. We should pay more attention to strengthen genuine qi to eliminate pathogenic factors in treatment of CHB.

Adolescent ; Adult ; Aged ; DNA, Viral ; blood ; Diagnosis, Differential ; Female ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; pathology ; therapy ; virology ; Humans ; Leukocytes, Mononuclear ; virology ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Syndrome ; Young Adult

Adult ; Burkitt Lymphoma ; metabolism ; pathology ; CD3 Complex ; metabolism ; DNA Nucleotidylexotransferase ; metabolism ; Diagnosis, Differential ; Female ; Granulosa Cell Tumor ; metabolism ; pathology ; Humans ; Ki-67 Antigen ; metabolism ; Leukocyte Common Antigens ; metabolism ; Ovarian Neoplasms ; metabolism ; pathology ; surgery ; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma ; metabolism ; pathology ; surgery ; Sarcoma, Myeloid ; metabolism ; pathology

OBJECTIVETo explore the clinical value of ultrasonic surface localization in internal jugular vein catheterization.

METHODSTotally 150 patients with American Society of Anesthesiologists physical status I -III who were planning to receive elective surgeries were randomized into anatomical landmark group, ultrasonic surface positioning group, and ultrasound-guided group using computed random table, with 50 cases in each group. The right internal jugular vein catheterization was performed after tracheal intubation. In the anatomic landmark group, patients were punctured using surface marks through central approach. In ultrasonic surface positioning group and ultrasound-guided group, patients were punctured with ultrasonic localization and guidance through central approach. The relationship between internal jugular vein and carotid artery, the position of the needle into the vein, the success rate of puncture, the change times of puncture point, and the complications were recorded.

RESULTSUltrasound scan revealed that the relationship between the right internal jugular vein and the right common carotid artery could be divided into three types: parallel (12.7%), partial overlapping (69.3%), and complete overlapping (18.0%). The average "safety distance" of jugular vein puncture was (1.15 +/- 0.47) cm. The success rate of the first puncture attempt in ultrasonic surface positioning group and ultrasound-guided group were 78.0% and 82.0%, respectively, which was significantly higher than that in anatomic landmark group (22.0%) (P < 0.05), whereas the complication incidence in anatomic landmark group (12.0%) were significantly higher than those in ultrasonic surface positioning group (0) and ultrasound-guided group (0) (P < 0.05).

CONCLUSIONSUltrasonic surface positioning applied during internal jugular vein catheterization is helpful to reveal the inner diameters as well as the origin and course of arteries and veins in the puncture and identify the abnormalities as early as possible. As a simple support technique for internal jugular vein puncture, it is suitable for clinical application.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Catheterization, Central Venous ; methods ; Female ; Humans ; Jugular Veins ; diagnostic imaging ; Male ; Middle Aged ; Ultrasonography ; Young Adult

Aged ; Female ; Humans ; Jaundice ; complications ; Liver Cirrhosis ; etiology ; Liver Diseases ; complications ; Sarcoidosis ; complications

OBJECTIVETo study the relationship between HBeAg expression and HBV-DNA in serum and peripheral blood mononuclear cells (PBMCs).

METHODS208 patients with chronic hepatitis B were included in this present study. HBV-DNA in the PBMCs were performed by polymerase chain reaction (PCR), with the serum HBV-DNA level being determined by the way of fluoresces quantities PCR (FQ-PCR). Meanwhile, HBV-GM was also detected via enzyme-linked immunosorbent assay (ELISA).

RESULTSThere were 106 patients for positivity in the HBV-DNA level of PBMCs with 102 for negativity, in which the HBV-DNA high levels (HBV DNA load > or = 1.0E5) in serum were 91.5%, 45.1% (chi2=52.12, P>0.01) respectively, with 76.4% and 50.9% (chi2=21.55, P>0.01) for the positive percentage of HBeAg expression.

CONCLUSIONA significantly positive correlation was found between HBV-DNA in PBMCs and serum HBV-DNA along with the positive percentage of HBeAg, indicating that obvious PBMCs' increase infected by HBV in patients with positivity of HBeAg and high level of serum HBV-DNA.

Adolescent ; Adult ; Aged ; Antibodies, Viral ; blood ; DNA, Viral ; blood ; genetics ; Enzyme-Linked Immunosorbent Assay ; Female ; Hepatitis B ; genetics ; immunology ; Hepatitis B e Antigens ; immunology ; Hepatitis B, Chronic ; blood ; virology ; Humans ; Leukocytes, Mononuclear ; virology ; Male ; Middle Aged ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Young Adult

Hybridoma cells were cultured by continuous perfusion in Fibra-Cel of 5L packed-bed bioreactor for 22 days in low serum or serum-free media. The corresponded amino acids were fed and serum concentration was decreased by analyzing glucose concentration, oxygen uptake rate, secretary antibody amount and amino acids concentration in culture supernatant. Comparing with continuous perfusion culture that amino acids were not fed, antibody amount of production was increased about 2-3 times. The inoculated cell density was 2.5 x 10(5) cells/mL, while the final cell density was 8.79 x 10(8) cells/mL. Antibody production was reached 295 mg/L/d at average level, and the highest level was reached 532 mg/L/d. These results provided a primary mode of enlarge culture for monoclonal antibody industralization.
Animals ; Antibodies, Monoclonal ; biosynthesis ; Cell Division ; Cells, Cultured ; Glucose ; pharmacology ; Hybridomas ; metabolism ; Mice ; Mice, Inbred BALB C ; Oxygen Consumption ; Perfusion

Coronary Artery Bypass ; adverse effects ; Female ; Humans ; Male ; Mammary Arteries ; injuries

Severe fever with thrombocytopenia syndrome bunyavirus is a newly emerging virus in China, enveloped with a tripartite, single-stranded RNA genome of negative polarity. The regulatory elements for viral transcription and replication, as well as encapsidation and packaging signals, are thought to be located within these noncoding regions (NCRs). The terminal nucleotides are genus specific and highly conserved. The function of the remaining nucleotides of the NCRs is still not well understood. In this study, we developed the plasmid-driven RNA polymerase I minireplicon system for SFTSV firstly, using reporter genes GFP and luciferase. The function of the noncoding regions of the three Bunyaviridae RNA segments (L, M, S) in transcription was analyzed. Reporter genes are successfully expressed in SFTSV minireplicon system. Our results suggest that the NCRs of SFTSV from all three segments contain the necessary signals to initiate transcription. Quantitative detection of the luciferase expression level shows that promoter activity in the three segments is different.
Bunyaviridae Infections ; virology ; Cloning, Molecular ; Genome, Viral ; Humans ; Phlebovirus ; genetics ; physiology ; Replicon ; Viral Proteins ; genetics ; metabolism ; Virus Replication

OBJECTIVESTo construct a stable HCV-producing cell model for anti-HCV drug research.

METHODSThe HCV-ribozyme recombinant plasmid pJFH1-Rbz was constructed to generate the exact 5' and 3' ends of HCV genomic RNA by placing two self-cleaving ribozymes at both ends of the HCV JFH-1 cDNA. The plasmid was then transfected into HepG2 cells and the resultant clones were screened with G418. Subsequently, immunofluorescence and Western blot were performed to detect the expression of HCV core protein, HCV RNA level was quantitated by TaqMan real-time PCR method and HCV particles was detected by electron microscopy.

RESULTSHCV core protein was detected in the screened cell clone, and the level of HCV RNA was up to 1000,0000 copies/ml in the culture medium. Electron microscopy showed the viral particles in the culture suspension were approximately 55 nm in diameter. IFN-treating experiment demonstrated that the HCV RNA level decreased with the increasing concentration of IFN alpha.

CONCLUSIONWe constructed a stable HCV-producing cell model which can be used for anti-HCV drug research.

DNA, Complementary ; Genome, Viral ; Hep G2 Cells ; Hepacivirus ; genetics ; Humans ; Plasmids ; RNA, Catalytic ; genetics ; Transfection ; Viral Core Proteins ; genetics ; Virion ; Virus Replication

OBJECTIVETo investigate the effects of different positive end expiratory pressures (PEEP) on functional hemodynamic parameters in patients lying in prone position during operation under general anesthesia.

METHODSTotally 60 patients undergoing cervical vertebra operation or lumbar vertebra operation were studied. All patients were also monitored with Vigileo／FloTrac system. The functional hemodynamic parameters including stroke volume variation (SVV), pulse pressure variation (PPV), and pleth variability index (PVI) under PEEP levels of 0 mmHg, 5 mmHg, 10 mmHg, and 15 mmHg were recorded before and after volume expansion (hydroxyethyl starch 6%,7 ml/kg). Fluid responsiveness was defined as an increase in stroke volume index (SVI) ≥ 15%(△SVI ≥ 15%). Responders were defined as patients demonstrating an increase in SVI ≥ 15% after intravascular volume expansion and non-responders as patients whose SVI changed <15%. Receiver operating characteristic (ROC) curves were generated for SVV, PPV, and PVI under different PEEP levels to determine their diagnosis accuracies and thresholds and their potential correlations.

RESULTSIn the prone position, SVV, PPV, and PVI were significantly higher compared to those in the supine position (P<0.05) and the mean arterial pressure significantly decreased (P<0.05); however, the changes of heart rate, stroke volume, SVI, cardiac output, and cardiac index showed no significant difference (P>0.05). In the prone position, along with the elevation of PEEP (0 mmHg, 5 mmHg, 10 mmHg, and 15 mmHg), the areas under the ROC curves of SVV were 0.864, 0.759, 0.718, and 0.521, the area under the ROC of PPV were 0.873, 0.792,0.705, and 0.505, and the area under the ROC of PVI were 0.851, 0.765 ,0.709, and 0.512. Under PEEP=0 mmHg, the diagnostic thresholds of SVV, PPV, and PVI were 10.5, 11.5, and 13.5. Under PEEP=5 mmHg, the diagnostic thresholds of SVV,PPV, and PVI were 11.5,13.5, and 14.5.Under PEEP=10 mmHg,the diagnostic thresholds of SVV, PPV, and PVI were 13.5,14.5, and 16.5.In the prone position,there was a significant correlation between SVV,PPV,PVI,and PEEP.

CONCLUSIONSSVV,PPV and PVI can predict fluid responsiveness similarly under the PEEP levels of 0,5, and 10 mmHg. Their diagnostic thresholds increases with the PEEP and the diagnostic accuracies decrease with the PEEP. However, under the PEEP level of 15 mmHg, SVV, PPV, and PVI can not predict fluid responsiveness accurately.

Anesthesia, General ; Blood Pressure ; Cardiac Output ; Hemodynamics ; Humans ; Positive-Pressure Respiration ; Prone Position ; ROC Curve ; Stroke ; Stroke Volume