OBJECTIVETo establish a set of suitable and reliable methods for HBV genotyping and to study the distribution of HBV genotypes.

METHODSType-specific nucleotides were searched through alignment of S genes (more than 1000 sequences) listed in GenBank. Then, type-specific primers were designed and type-specific primer PCR was used to genotype the 238 HBV strains. S genes of the untyped strains were further amplified and sequenced to find out their genotypes with type-specific nucleotide analysis.

RESULTSAll the 238 HBV strains were genotyped. 159 (66.8%) cases were genotype B, 69 (28.9%) were genotype C, 6 (2.5%) were mixtures of genotypes B and C and 4 (1.6%) were mixtures of genotypes B and D. No genotypes of A, E, F, G, and H were found.

CONCLUSIONGenotypes B and C are the most common types for HBV strains. Mixtures of genotypes B and C or genotypes B and D coinfection rarely existed. There is no relationship between the gender of the patients and HBV genotypes (X2 = 0.794, P more than 0.05).

DNA Primers ; DNA, Viral ; blood ; genetics ; Female ; Genotype ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; virology ; Humans ; Male ; Nucleotides ; genetics ; Polymerase Chain Reaction ; methods ; Sequence Analysis, DNA

Objective To evaluate the value of three-dimensional contrast-enhanced angiography (3D CE MRA) and MRI in the diagnosis and delineation of cerebral arteriovenous malformation(AVM). Methods Twenty-two cases of cerebral AVM examined by MRI and 3D CE,MRA.DSA was performed in 17 cases.A three-dimensional fast low angle shot (3D FLASH) was used for 3D CE MRA with Gd-DTPA dosage of 0.2 mmol per kilogram for body weight.The source images were subtracted from mask images and transferred to computer workstation using three-dimensional reconstruction.Results Among 22 cases, 19 showed typical AVM“flow void”signal on MR images.3D CE MRA clearly displayed the nidus,feeding artery and draining vein.All of the foci were above the cerebellum tentorium.13 located within one lobe, 3 exceeded one lobe and 3 situated in the deep of cerebrum.Feeding arteries were derived from single artery in 9 cases,and mixed supply in 10 cases.Draining veins diverted to sagittal sinus and/or sigmoid sinus in 6,deep cerebral veins in 8 and mixed in 5.In the 14 cases examined by both DSA and 3D CE MRA,3D CE MRA was superior to DSA in three-dimensional demonstration of the nidus,but inferior to DSA in demonstration of some details.3D CE MRA depicted 78.4% feeding arteries and 84.0% draining veins in addition,tiny pathologic blood vessels smaller than 1 centimeter were detected by DSA,but could not be found by 3D CE MRA and were only shown as hemorrhage lesions on MR images in 3 cases.Conclusion As a non-invasive technique,3D CE MRA combined with MRI is accurate in diagnosis and localization of cerebral AVM,and should be used as the first choice for those clinically suspected of AVM.But DSA remains needed for demonstration of details and tiny AVM.

BACKGROUND:The immunomodulatory ability of bone marrow mesenchymal stem cells(BMSCs)gives it a promising future in treating graft-versus-host disease(GVHD),especially with previous success in treating patients with acute GVHD.However,there are fewer reports concerning BMSCs in treating chronic GVHD,particularly for sclerodermatous chronic graft-versus-host disease(ScGVHD).OBJECTIVE:To evaluate the efficacy and safety of treatment of BMSCs for ScGVHD,and to primarily explore the immunological mechanism of clinical efficacy.METHODS:Four ScGVHD patients at the Affiliated Hospital of Academy of Military Medicine Science,between September 2006 and August 2008,were enrolled for this trial.The median patient age was 41 years,1 female and 3 male.The patients received BMSCs infusion at a dose of(1.0～2.0)×10~7 cells every time by intrabone marrow injection from the anterosuperior iliac spine and BMSCs from the same donor for the same patient were infused more than once.Concomitant medications for ScGVHD were individualized for each patient,but all were current standard medicines and the doses were significantly tapered.RESULTS AND CONCLUTION:After BMSCs infusion,the ratio of Th1 to Th2 was dramatically overturned,with an increase of Th1 and a decrease of Th2 reaching at a new balance.Correspondingly,symptoms of all the four patients gradually improved.During the course of BMSCs treatment,the life signs and laboratory results from the recipients remained normal.By the time of this report,there has been no recurrence of leukemia in the four patients.Although this study alone cannot guarantee the application of BMSCs in ScGVHD,the results are strongly in favor of the idea that the BMSCs treatment for ScGVHD patients is therapeutically practical without any detectable side effects,which may provide a new insight into the matter of treating ScGVHD clinically,thus will greatly increase the survival rate of leukemia after allogeneic bone marrow transplantation.

Through analysis and summary of the biological characteristics of various biomarkers, to explore the reliability of different markers for early diagnosis of osteoarthritis. Cartilage oligomeric matrix protein, N-terminal crosslinked telopeptide of type Ⅰ collagen and C-terminal crosslinked telopeptide of type Ⅱ collagen are the possible effective markers in osteoarthritis early diagnosis. Hyaluronic acid and C-terminal crosslinked telopeptide of type I collagen are more suitable for evaluating the ollurrence and derelopment of osteoarthritis. The efficacy of miRNA and lncRNA in osteoarthritis diagnosis and evaluation remains to be proved. Each marker may has two or more biological effect, this paper will focus on finding out an accurate and stable marker with the analysis and summary of present bio-markers.

OBJECTIVE To investigate the underlyingmechanism on the association of red blood cell and gut microbiota in rats induced by High-Fat Diet(HFD).METHODS A total of 36 male Sprague-Dawley rats (180±20g) were randomly divided into two groups. The control group (n=10) was given a normal chow diet(10％ calories of fat),and the High-fat diet group(n=26)was given a HFD(60％ calo-ries of fat).We recorded body weight,length and detected serum glucose,serum lipids and insulin ev-ery two weeks.The fresh arterial blood was collected during the experiments and blood gases were measured immediately (Radiometer Medical ApS, Denmark).Thehematocrit (Hct) and partial pressure of oxygen(pO2)were detected by the sensor cassette,following themanufacturer′s instructions.The de-tection method was conductivity measurements and current method, respectively. The feces from ce-cum were analyzed by 16S rRNA gene high-throughput sequencing(Illumina Miseq,USA). RESULTS According to the insulin resistance(IR),body weight and body length,the model group was divided into two small groups.(1)IR group,in which IR,body weight and body length were higher than the control group (P<0.05). (2) un-IR group, body weight and body length were higher than the control group (P<0.05),but the IR was not significantly different.In addition,the levels of hematocrit(Hct),checktotalhe-moglobin (ctHb) and check total blood oxygen content (ctO2) showed significantly increased in the IR group when compared with the control group (P<0.05), however, the pO2was not statistically signifi-cant. Furthermore, we identified that the genus Lactobacillus was moderate positive correlation with Hct,ctHb and ctO2(P<0.05).Compared with the control group,the relative abundance of the Lactoba-cillus was significantly lower in IR group(P<0.05).CONCLUSION The high-fat diet induced rats′local tissue hypoxia under the red blood cell increasing,oxygen partial pressure constant and the reduction of Lactobacillus′abundance might be caused by aerobic oxidation and glycolysis inhibition in the meantime.

OBJECTIVETo explore the toxic effect of fluoride on the human thyroid cells (Nthy-ori 3-1) and its mechanism.

METHODSNthy-ori 3-1 cells were exposed to 0.0, 0.1, 1.0, 3.0 mmol/L of sodium fluoride (NaF) in vitro. After 24 hours incubation, 3 (4,5-Dimethylthiazol-z-yl)-3, 5-diphenyltetrazolium bromide (MTT) assay and lactate dehydrogenase (LDH) assay were used to measure cell viability and the LDH leakage rate. Reactive oxygen species (ROS) level, constituent ratio of the cell cycle, and apoptosis rate were measured by flow cytometry.

RESULTSComparing to viability of control group (set as 100.00%), the cell viability of the 1.0, 3.0 mmol/L fluoride-treated groups (76.64 +/- 9.13)%, (64.04 +/- 6.32)% were significantly decreased (all P values <0.01). LDH leakage rate and ROS level of the 3.0 mmol/L fluoride-treated group ((48.66 +/-7.15)%, (29993.50 +/- 1786. 86) FI) were significantly increased (all P values <0.01) compared to control group ((35.24 +/- 3.02)%, (13021.33 +/- 1067.55) FI). The G0/G1 phase cells of the 1.0 mmol/L fluoride-treated group ((40.76 +/- 5.65)%) were lower than control group (60.09 +/- 1.76)% (P < 0.01), yet the percentage of cells in S phase ((54.05 +/- 4.59)%) were higher than the control group (32.59 +/- 2.43) % (P < 0.01). Comparing to control group ((9.64 +/- 3.44)%), the percentage of apoptosis cells increased in the 3.0 mmol/L fluoride-treated group ((20.09 +/- 3.22)%) (P < 0.01).

CONCLUSIONTo Nthy-ori 3-1 cells, fluoride under experimental concentrations decreases cell viability, improve the LDH leakage rate, and ROS level. It blocks the cells in S phase and induce cell apoptosis.

Apoptosis ; drug effects ; Cell Cycle ; Cell Division ; Cell Line ; Fluorides ; toxicity ; Humans ; Reactive Oxygen Species ; analysis ; Thyroid Gland ; cytology ; drug effects

Objective To compare the applicability of different occupational health risk assessment methods in small furniture manufacturing industry. Methods American EPA inhalation risk model, Singapore semi-quantitative risk assessment model of occupational exposure to chemical substances and Australian Occupational Health and Safety Risk Assessment model, were used to assess the occupational health risk in a small furniture manufacturing industry from a city of Zhejiang Province. Results The results of American EPA model showed that the workers who exposed to benzene and formaldehyde had low risk of carcinogens, and who exposed to benzene and xylene had very high risk of non-carcinogens. According to Singapore semi-quantitative risk assessment model, there were low and medium health risk caused by toluene and xylene, and high risk caused by wood dust in preparation and polishing jobs. Similar to the results of other models, Australia qualitative risk assessment model showed that there were medium health risk caused by toluene and xylene, and high risk caused by benzene, wood dust and noise. All of the three methods could found the key risk control point in furniture manufacturing industry. The risk ratios of the three methods were higher than the toxic work classification ratio (P<0.01), and the risk ratio of EPA model were higher than the results of Singapore model and Australia model (P<0.05) . Conclusion All of the three methods can be applied to assess the occupational health risk in furniture manufacturing industry, and the combined application of multiple risk assessment methods can be used as one of the risk assessment strategies.

Objective:To investigate the effects of posterior condylar offset (PCO) change on functional recovery after high-flexion posterior-stabilized total knee arthroplasty (TKA).Methods:From December 2018 to May 2019, a total of 76 patients (7 males and 69 females) who underwent primary TKA were included. The age of patients was 67.78±5.13 years (56-75 years). Preoperative and postoperative radiological PCO were measured by lateral knee X-ray. The true preoperative PCO was defined as the sum of radiological PCO and the thickness of posterior condylar cartilage. According to the changing of PCO (ture preoperative PCO - postoperative PCO), the subjects were divided into four groups, namely 28 cases in ≤-3 mm group, 23 cases in -3 mm- group, 15 cases in 0 mm- group and 10 cases in ≥3 mm group. The parameters, including age, body mass index, range of motion (ROM), Knee Society Scores (KSS) and visual analogue score (VAS) before the operation, were not significantly different among four groups. ROM, KSS, VAS at 2 weeks, 1 month, 3 months, 6 months after the operation were compared among the four groups.Results:There were good inter-observer reliabilities regarding the parameters measured in this study ( ICC>0.75). The KSS, ROM, VAS of all the subjects after operation were significantly better than those before the operation ( F=318.768, 64.983, 361.749; P=0.000). In all groups, the recovery of KSS and VAS last to 6 months after the operation. The ROM trended to be stable at the 3 months after the operation. At 6 months after operation, ROM, KSS and VAS of ≤-3 mm group was 116.07°±9.66°, 156.25±21.49, and 1.18±0.94, respectively. These parameters of -3 mm- group was 119.57°±7.52°, 162.17±17.09, and 1.26±0.86. However, these parameters of 0 mm- group was 126.07°±5.25°, 161.86±8.86, 1.00±0.55, respectively. These of ≥3 mm group was 118.00°±4.21°, 156.60±16.98 and 1.30±0.95. The KSS, KSS anatomy score, KSS function score and VAS were not significantly different at any follow-up point among four groups. The ROM of 0 mm- group at 1 month, 3 months and 6 months after operation (118.57°±13.07°, 25.00°±6.20°, 126.07°±5.25°) was significantly different from other three groups ( F=4.966, P=0.003; F=4.179, P=0.006; F=5.262, P=0.003), while 0 mm- group's ROM were greater than other three groups ( P<0.05). Conclusion:Increasing within 3 mm of PCO was related to larger postoperative ROM in high-flexion posterior-stabilized TKA. However, change of PCO had no influence on the outcomes of KSS recovery and pain relief.

OBJECTIVETo study the method for synthesis of 2-hydroxyl-5- butyramidobenzoic acid and test its effect on acetic acid-induced colitis in rats.

METHODS2-hydroxyl-5-butyramidobenzoic acid was synthesized from 5-aminosalicylic acid and butyric acid by amidation, esterification and hydrolization. The effect of 2-hydroxyl-5-butyramidobenzoic acid on acetic acid enema-induced colitis in rats was investigated.

RESULTSThe structure of 2-hydroxyl-5-butyramidobenzoic acid was identified by IR and 1H-NMR. After treatment with acetic acid, the colon mucosal damage index (CMDI), fecal occult blood (OB) test, and activity of myelperoxidase (MPO) increased significantly in the rats as compared to the control levels. 2-hydroxyl-5- butyramidobenzoic acid obviously reduced the CMDI and OB, and reduced the level of MPO in the rats with colitis.

CONCLUSIONThe synthesis of 2-hydroxyl-5-butyramidobenzoic acid requires only mild conditions with simple procedures, and the synthesized 2-hydroxyl-5-butyramidobenzoic acid shows obvious therapeutic effects on mucosal damage of in rats with acetic acid-induced colitis.

Acetic Acid ; Aminobenzoates ; chemical synthesis ; chemistry ; pharmacology ; therapeutic use ; Animals ; Colitis, Ulcerative ; chemically induced ; drug therapy ; Male ; Protective Agents ; chemical synthesis ; pharmacology ; therapeutic use ; Rats ; Rats, Sprague-Dawley ; Salicylates

OBJECTIVETo study natural history of small hepatocellular carcinoma (sHCC) and value of MRI in this research.

METHODSFrom 1999 to 2002, serial MR imagings were performed in 192 patients with cirrhosis no less than twice in order to detect sHCC focus. Characteristic MRI findings of focus were analysed. Diameters of focus (D) were measured. Tumorigenesis detected time (T) was counted; T and D were analysed by statistics and tumor volume doubling time (T(d)) was calculated.

RESULTSFifty-eight new tumor foci in 33 patients were found, including 31 foci in 23 patients developed on the basis of cirrhosis, with a 12.0% cumulative tumorigenic rate of three years. The tumorigenesis detected time (T) was 386.9 +/- 256.4 days and the maximum diameter (D) was 2.258 +/- 1.074 cm. T was statistical correlated with D(3) (P < 0.01) and regressive formulas were established: D(3) = -2.69 + 0.058T (P = 0.0007), D(3) = -72.13 + 16.04lnT (P = 0.0064). Tumor volume doubling time was 20 approximately 279 days (mean: 104 days).

CONCLUSIONThe percentage of tumorigenesis is high in patients with cirrhosis. Regular MRI examinations are necessary to detect sHCC early. The optimal interval time is 3 approximately 6 months.

Adult ; Aged ; Carcinoma, Hepatocellular ; diagnosis ; Female ; Follow-Up Studies ; Humans ; Liver Cirrhosis ; pathology ; Liver Neoplasms ; diagnosis ; Magnetic Resonance Imaging ; Male ; Middle Aged