Objective To isolate HIV-specific T cell clone and to expand them in vitro through the activation-induced expression of CD137 molecule.Methods The peripheral blood mononuclear cells were isolated from HIV-infected patients and HIV Gag specific CD3+CD8+CD137+T cell subset were sorted to 96-well plate in 1 cell/well by multicolor flowcytometry and single cell sorting.After 14 days in vitro culture with feeder cells and cytokines, the numbers and phenotypes of the cultured HIV-specific T cells were calcu-lated and identified.Results The CD137 expression was low on rested T cells but up regulated by the stim-ulation with Gag peptide pool.The CD8+CD137+T cells could secret IFN-γ.The number of CD8 T cells reached to 106 after 14 days in culture and expanded to 107-108 cells after 28 days of culture in vitro 100%of the cells remained activated upon Gag stimulation.Conclusion In stead of using IFN-γ, CD137 could be utilized as a novel molecule to isolate and expand HIV specific T cells in vitro.The expanded antigen spe-cific T cell clones could maintain good activation status.

Objective:To establish the quality standard for Tangzhixiao capsules. Methods: The qualitative identification of Salvia miltiorrhiza,Atractylodes and Hawthorn was detected by TLC, and the quantitative determination of salvianolic acid B was determined by HPLC. The HPLC determination was performed on a YMC-Triart C18 (250 mm × 4. 6 mm, 5μm ) column with the mobile phase consisted of acetonitrile-0. 1% phosphoric acid (24∶ 76) at the flow rate of 1. 0 ml·min-1 , the column temperature was 30℃ and the detection wavelength was set at 286 nm. Results: The TLC spots were fairly clear, and the negative samples showed no interference. The concentration of salvianolic acid B within the range of 0. 012-0. 120 mg·ml -1 was linear with peak area(r = 0. 999 5), the average recovery was 100. 06% and RSD was 0. 83%(n = 6). Conclusion: The method is accurate, reliable and reproducible, which can be used in the quality control of Tangzhixiao capsules.

Objective To analyze the complications after procedure for prolapse and hemorrhoids (PPH) in treating severe mucocutaneous hemorrhoids.Methods The clinical data of 550 patients with severe mucocutaneous hemorrhoids who were admitted to the Hangzhou Hospital of Nanjing Medical University from January 2005 to December 2013 were retrospectively analyzed.All the patients were treated by PPH or PPH + external hemorrhoids resection.The postoperative complications were recorded and assessed.Patients were followed up via outpatient examination and telephone interview till June 2014.Results Operation was successfully carried out on all the 550 patients.The operation time and duration of hospital stay were (15 ± 10)minutes and (4.2 ± 1.8)days,respectively.A total of 445 patients were followed up for 6-60 months,with the median time of 28 months.The overall incidence of complications was 14.61％ (65/445).Forty-five patients were complicated with anal pain,12 with hemorrhoid recurrence,7 with bleeding,5 with dysporia,4 with anastomotic stenosis and 2 with rectal and perianal infection.Of the 12 patients with hemorrhoids recurrence,5 was alleviated by medication and 7 received PPH for the second time.Of the 4 patients with anastomotic stricture,the condition of 2 patients were alleviated by anus dilatation,and the condition of 1 patient was alleviated by balloon dilatation under enteroscope,1 patient received circular incision of the stricture because of severe stricture and failed of conservative treatment.Of the 2 patients with rectal and perianal infection,1 was alleviated by intravenous and local medication,and the other patient who was complicated with anal fistula was alleviated by surgery.The condition of other patients was improved after symptomatic treatment.Conclusions PPH is effective and safe for the treatment of severe mucocutaneous hemorrhoids.The main complication is anal pain,and it could be cured or alleviated by symptomatic treatment.

The secondary structure of the protein of DM0 and DMT in Oreochromis aureus were predicted by the methods of Garnier-Robson, Chou-Fasman and Karplus-Schulz based on the amimo acid sequences of DM0 and DMT. And Hydrophilicity plot, Surface probability and Antigenic index for DM0 and DMT protein were obtained by the methods of Kyte-Doolittle, Emini and Jameson-Wolf, respectively. Combined the results according to these methods, the B cell epitopes for DM0 and DMT protein were predicted. The results demonstrated that there were some centers of?-helix in the DM0 protein' s N- terminal No. 80 - 112, 144 -147, 193- 194, 251 - 255, 260 - 269 and No. 279- 283, and in the DMT protein' s N-terminal No. 61 -86, 98 - 105, 140 - 146, 239 -241 and No. 269 -273. And there are some centers of?-sheet in the DM0 protein' s N-terminal No. 59 -61, 69 -70, 148 - 150 and No. 383 -390, and in the DMT protein's N-terminal No. 125 -129, 207-213, 255-264 and No. 281-284. Furthermore, the DMO protein' s N-terminal No. 40-41,44 -45, 50-51, 128-129, 189-192, 204-207, 216-222, 226-233, 244-246, 298 - 299 and No. 323 -326, and the DMT protein' s N-termianl No. 12 - 13, 26 - 27, 43 - 44, 58 - 60, 93 - 95, 115 - 120, 136 -139 and No. 149 -151 may be the flexible regions. Moreover the B-cell epitopes possibly localized in or nearby the DMO protein's 1 -5, 41 -51, 65-67, 86-89, 98-110, 154-170, 183-203, 205 -248, 258-264, 284 - 291, 293 - 298, 270 - 375, 389 - 392 and No. 402 - 410, and DMT protein' s N-termianl No. 1 - 9, 17 - 28, 77 - 84, 114 - 123 , 131 - 139, 157 - 184 and No. 96 - 207. Theses results are helpful for studies on sex control mechanism of DMO and DMT in Oreochromis aureus.

OBJECTIVETo study the influence of experimentally osteoporosis to osteogenic efficiency of bone matrix gelatin(BMG) implanted into the calvarial defects of rats.

METHODSSixty-eight female SD rats of 12 +/- 1 weeks were randomly divided into two groups with 34 rats in each group. The ovaries were excised in the ovariectomized group (VG). The control group underwent sham surgery. Ninety days after ovariectomy, 10 rats from each group were examined to ensure the formation of postmenopausal osteoporosis by measuring bone density of the femur with single photon absorptiometric measurements. A critical-sized (8 mm in diameter) calvarial defect was created on the rest of 48 rats. Bone matrix gelatin was implanted to the defect. The rats were scarified at the 21st and 56th day after surgery respectively. The new bone forming capability of BMG was evaluated with undecalcified histological observation, tetracycline fluorescence marker, quantitative bone histomorphometry, At 90th day after ovariectomy, bone density of scanning electron microscopy and X-ray spectrometry.

RESULTSOVG showed very significant difference compared with the control group (0.315 +/-.015) g/cm2 vs [(0.347 +/- 0.017) g/cm2, P < 0.01 ]. At the 21st day following the implantation operation, new bone formed within the bone defects in both groups. The amount of new bone in OVG was lower than the control group. The tetracycline-labeled region in the bone defect was sparser in the OVG. At the 56th day, the bone defects healed mostly in the control group but fibrous tissue filled parts of bone defect in the OVG. The distance between two fluorescent lines of incorporated tetracycline and the mean mineralization deposition were significantly lower in the OVG than the control at the 21st day and 56th day. Mineralization of callus in OVG was inferior. Significant difference was found between the OVG and the control group in the calcium to phosphate ratio of callus in bone defects at the two time-points.

CONCLUSIONSExperimentally induced osteoporosis depressed osteogenic efficiency of BMG, suggesting that estrogen could play an important role in bone remodeling with bone substitute participating.

Animals ; Bone Matrix ; chemistry ; Bone Regeneration ; Female ; Femur ; metabolism ; Gelatin ; therapeutic use ; Osteogenesis ; Osteoporosis ; surgery ; Ovariectomy ; Rats ; Rats, Sprague-Dawley ; Skull Fractures ; surgery

OBJECTIVETo investigate the etiology of 1977 patients from northern China with acute (ALF), sub-acute (SALF) or acute-on-chronic liver (ACLF) failures.

METHODThe age, gender, etiology, pathogenesis, and prognosis of the 1977 patients with liver failures were retrospectively analyzed.

RESULTSOf the 1977 cases, the three most common causes of ALF were HEV (33.96%) or HBV (13.21%) infections or those caused by medicines (9.43%). The three predominant causes of SALF were medicines (31.53%), HEV (16.22%) or HBV (9.91%) infections, but those of the ACLF were HBV (90.29%) infection, alcoholic hepatopathy (2.65%), and HBV super infected with HEV (2.26%) infections. 90.09% (1781) patients were infected by hepatotropic viruses. Of these 1781 patients, the most common cause of their liver failures was HBV infection (92.93%). In these HBV infected patients, 77.10% were from 26 to 55 years old. From 2005 to 2007, there were 39 patients with alcoholic liver failure. In the past two years, there were 23 patients with drug induced liver failure. The improvement rate of the 1977 patients after their treatments was 35.56%. The improvement rate of HEV infected liver failure was higher than drug induced liver failure (P less than 0.05); no statistical significance was found between other groups (P more than 0.05).

CONCLUSIONDifferent types of liver failure have different predominant causes. HBV infection is the most common cause in our 1977 patients. In the past two years, the number of drug induced liver failures and alcoholic liver failures have been increasing.

Acute Disease ; Adult ; Chemical and Drug Induced Liver Injury, Chronic ; etiology ; Chronic Disease ; Female ; Hepatitis B ; complications ; Hepatitis E ; complications ; Humans ; Liver Diseases, Alcoholic ; etiology ; Liver Failure ; chemically induced ; classification ; etiology ; virology ; Male ; Middle Aged ; Prognosis ; Retrospective Studies

OBJECTIVETo seek the effective treatment for peripheral neuropathy induced by chemotherapeutic drugs.

METHODSSixty-four cases of peripheral neuropathy induced by Paclitaxel or Oxaliplatin were randomly divided into an acupuncture group and a medication group, 32 cases in each group. The acupuncture group was treated with therapeutic principle of dredging meridians and collaterals, tonifying qi and eliminating blood stasis, supplementing liver and kidney, nourishing blood and tendon. Hegu (LI 4), Taichong (LR 3), Zusanli (ST 36), Qihai (CV 6) and Quchi (LI 11) etc. were selected. The medication group was treated with intramuscular injection of Cobamamide. The neurotoxicity of two groups was compared with questionnaire of peripheral neuropathy induced by chemotherapeutic drugs before and after treatment.

RESULTSThe total effective rate for sensory nerve disorder of acupuncture group was 66.7% (20/30), which was superior to that of 40.0% (12/30) in medication group (P < 0.05).

CONCLUSIONAcupuncture is more effective than Cobamamide for treatment of peripheral neuropathy induced by chemotherapeutic drugs, especially for moderate and severe sensory nerve disorder induced by paclitaxel.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Drug-Related Side Effects and Adverse Reactions ; Female ; Humans ; Male ; Middle Aged ; Organoplatinum Compounds ; adverse effects ; Paclitaxel ; adverse effects ; Peripheral Nervous System Diseases ; chemically induced ; therapy

OBJECTIVETo evaluate the clinical effect of fluoride varnish in preventing caries of primary teeth, and to compare the caries prevention effects of fluoride varnish with two different concentrations.

METHODS150 children whose ages were three year-old were randomly chosen from one of nurseries of Chongqing, and were randomly divided into four groups with 37 children in 0.5% fluoride varnish group, 38 children in 0.1% fluoride varnish group, 39 children in 0.5% sodium fluoride group and 36 children in blank group. The groups included 71 'girls and 79 boys. Protective measure was applied every half an year and the research has lasted for two years. The research was designed by double-blinded methods, and was carried out with the dental caries standard suggested by World Health Organization. All the data were collected and the clinical effects of four groups were compared.

RESULTSThe average caries index and caries surface index of 0.1% fluoride varnish group and those of control groups were significantly higher than that of 0.5% fluoride varnish group. There were no significant difference between the average caries index and caries surface index of 0.1% fluoride varnish group and those of control groups.

CONCLUSION0.5% fluoride varnish was effective in preventing caries of primary teeth.

Cariostatic Agents ; Child ; Dental Caries ; Female ; Fluorides ; Fluorides, Topical ; Humans ; Male ; Paint ; Phosphates ; Sodium Fluoride ; Tooth, Deciduous

Female ; Hepatitis, Viral, Human ; therapy ; Humans ; Liver, Artificial ; Male ; Plasma Exchange ; methods ; Treatment Outcome

OBJECTIVETo profile the protein expression in activated rat hepatic stellate cells (HSCs).

METHODSPrimary rat HSCs were isolated and cultured in vitro. After 10 days in vitro culture, the HSCs were activated. Total protein extracted from these activated HSCs were digested, and the obtained peptides were analyzed by using online 2D nanoLC-MS/MS. The identified proteins were classified according to their distributions and functions.

RESULTS1014 proteins were identified from 50 microg HSCs protein extract, the molecular weights of these proteins ranged from 7832 Da to 588,364 Da. Most of these proteins resided in nucleus, cytoskeleton, mitochondrion and endoplasmic reticulum. And these proteins were mainly involved in nucleic acid metabolism, organelle organization, signal transduction and energy generation. Among these proteins, alpha-smooth muscle actin, vimentin and desmin were specifically expressed in activated HSCs.

CONCLUSIONTo the best of our knowledge, this is the most comprehensive protein expression profile of activated rat HSCs.

Actins ; analysis ; metabolism ; Animals ; Cell Nucleus ; metabolism ; Cells, Cultured ; Chromatography, High Pressure Liquid ; methods ; Desmin ; analysis ; metabolism ; Hepatic Stellate Cells ; metabolism ; Male ; Proteome ; analysis ; metabolism ; Proteomics ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry ; Vimentin ; analysis ; metabolism