Objective To investigate the features of HIV-1-specific Cytotoxic T-Lymphocyte (CTL)responses in Chinese.Methods The HIV-1-specific CTL responses were analyzed in an IFN-?ELISPOT assay by using a matrix system containing 820 overlapping peptides spanning the entire HIV-1 Clade B and C consensus sequence.Results The HIV-1-specific CTL response almost clus- tered in Gag and Nef across either HIV-1 Clade B(HIV-1 B)or HIV-1 Clade C(HIV-1 C),while oth- er proteins could also be recognized at different level.In comparison of the response between HIV-1 Clade B and Clade C,the magnitude and frequency was roughly identical with some difference found at single-peptide level.The most frequently recognized peptides of HIV-1 B were located in Nef,GPKEP- FRDYVDRFYKTLR(5/17,29.4%)and Gag,LWVYHTQGYFPDWQNY(5/17,29.4%),while the most frequently recognized peptide of HIV-1 C was located in Gag GPKEPFRDYVDRFFKTLR(6/17, 35.29%).Conclusions HIV-1-specific CTL responses clustered within HIV-1 Gag and Nef in Chinese. However,there was some difference between HIV-1 B and HIV-1 C at single-peptide level.

Plants in Ainsliaea genus, belongs to Compositae family, are traditional Chinese medicine and widely used in folk. These plants contain various types of chemical components, and main components are sesquiterpene lactone and its glycosides. In addition, there are triterpenoids, flavonoids, steroids, phenolic acid, long chain fatty acid and volatile oils. Recently, much attention has been payed to varlous research of A. fragrans. This paper reviewed and summarized the chemical components to provide the theoretical basis for the use of Ainsliaea.
Asteraceae ; chemistry ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Molecular Structure

For in vitro studies, both CD34+ selected cell and mononuclear cell (MNC) can be used to expand hematopoietic stem/progenitor cells. To investigate the expansion characteristics of mononuclear cells (MNC) and CD34+ selected cells the two cell fractions were cultured in the medium containing cytokine cocktails of SCF + IL-3 + IL-6 + FL + Tpo. It was found that the CD34+ selected cells had presented a high proliferation potential. The expansion of CD34+ selected cells could be maintained for 8 weeks while that of MNCs declined after 4 weeks. During the culture period, the maximum expansion of total cells in CD34+ selected cell culture achieved 31,270.9 +/- 8640.5 times, while that of MNC reached 50.9 +/- 8.2 times only. In the culture of MNCs, the colony density and the proportion of CD34+ cells increased from day 0 to day 7. However, in the culture of CD34+ selected cells, both the colony density and the proportion of CD34+ cells declined continuously during the whole culture period. During the ex vivo culture of CD34+ selected cells, the maximum expansion of CFU-GM and CD34+ cells achieved 185.7 +/- 14.1 fold and 191.7 +/- 188.8 fold, respectively. They are much higher than that of MNC, which were 12.4 +/- 3.2 fold and 50.6 +/- 33.2 fold only. While the BFU-E of both cell fractions only expanded by few times, which were 7.2 +/- 5.2 and 10.1 +/- 3.4 times, respectively. The results showed that the CD34+ selected cells culture could obtain more CFU-GM cells and CD34+ cells during the whole culture period.
Antigens, CD34 ; analysis ; Cell Count ; Cell Separation ; Cells, Cultured ; Fetal Blood ; cytology ; Hematopoietic Stem Cells ; physiology ; Leukocytes, Mononuclear ; cytology

Objectivc To investigate the protective effect of simvastatin on retinal ganglion cells (RGC) after optic nerve crush (ONC).Methods Together 50 Kunming mice were randomly divided into normal group,sham group,ONC group and simvastatin protection group.The mice in the normal group were untreated,the sham group was treated with the exposure of the optic nerve without injury,the ONC group mice underwent ONC operation on the left eyes,followed by intravitreal administration of equilibrium solvent [50 mg · mL-1 ethanol plus 1 mol · L-1 NaOH,which were activated by 1 mol · L-1 HC1 (pH 7.2)],and the simvastatin protection group was intravitreally injected different concentrations of simvastatin (0.5 g · L-1,1.0 g · L-1,1.5 g · L-1) after ONC operation.Brn3a immunofluorescence staining,HE staining and toluidine blue staining were used to detect the apoptosis of RGC and pathological changes of optic nerve.Results On day 7 after operation,in the ONC group,the apoptosis of RGC was observed obviously,with the survival rate dropping to (35.1 ± 3.9) ％,and the thickness from the retinal ganglion cell layer to outer nuclear layer was decreased from (123.13 ± 1.04) μm to (97.48 ± 2.33) μm,which was significantly thinner than that in the control group (P ＜ 0.01);moreover,the fibrous bundle of optic nerve disappeared,and the neuroglial cells were clustered into groups,as well as the axons showed swelling and serious degeneration,but after intravitreal injection of 1.0 g · L-1 simvastatin,the survival rate of retinal ganglion cells increased to (76.3 ± 3.7) ％ (P ＜ 0.05),and the aforementioned thickness was increased to (111.39 ± 4.06) μm,which was statistically significant when compared with the ONC group (P ＜ 0.01).The degeneration of optic nerve was improved,and the structure of neuroglial cell axons and the nerve fibers became normal.Conclusion Simvastatin can reduce the optic nerve degeneration and improve the survival rote of retinal ganglion cells.

Stirred culture offers a number of advantages over static systems as it maintains a stable, homogeneous culture environment and is easy to scale-up. This paper focused on the development and application of stirred tank bioreactor to culture hematopoietic cells. Preliminary study of stirred culture of hematopoietic cells was carried out in cord blood mononuclear cells culture in spinner flask. The results showed that the amplification rates of total cell, CFU-GM and BFU-E, with the exception of CFU-Mk, were greater in spinner flask than T-flask. The number of total cells increased 20 fold after 14 days incubation in spinner flask. The amplification rates of CFU-GM, CFU-Mk and BFU-E reached maximum at 10th day, 10th day and 7th day respectively, and the maximal amplification rates were 9.2-fold, 5.5-fold and 2.4-fold respectively, whereas the rate of CD34+ cells in spinner flask was (6.7 +/- 4.0)-fold at day 10. These results indicated that the stirred culture system is better than the static culture systems for hematopoietic cell proliferation. The biocompatibility of cord blood MNC to different types of materials used in bioreactors was also tested. The results showed that glass, stainless steel 316L and polytetraflouroethylene (PTFE) supported the growth of hematopoietic cells well. A higher cell density was reached in stirred bioreactors with controlled pH and DO than static culture. These findings suggested that the controlled large-scale culture could be used to overcome the clinical shortage of hematopoietic cells.
Antigens, CD34 ; metabolism ; Bioreactors ; Cell Culture Techniques ; instrumentation ; methods ; Erythroid Precursor Cells ; cytology ; Fetal Blood ; cytology ; Granulocyte-Macrophage Progenitor Cells ; cytology ; Humans ; Polytetrafluoroethylene ; Stainless Steel

Clinical evidence of hematopoietic restoration with umbilical cord blood (UCB) grafts indicates the UCB can be a useful source of hematopoietic stem cells for routine bone marrow reconstitution. Considering (10 +/- 5) x 10(8) nucleared cells per cord blood unit, there is a potential limitation for the use of cord blood in adults, which, however, can be overcome by ex vivo expansion of cells. A prerequisite for expansion is the significantly higher recovery of MNC, CD34+ cells and colony-forming cells (CFC) by thawing cryopreserved MNC. Cooling rate always acts as a critical factor that can affect the recovery of cells. Although the rate of - 1 degrees C/min is adopted in most of the cryopreservations, no data has been reported about the detailed effects of different cooling rates. The aim of the study was to reveal the different effects of cooling rates on cryopreservation of hematopoietic stem cells from cord blood. UCB samples were collected, and cryopreserved as mononuclear cells (MNC) with different cooling rates of - 0.5 degrees C/min, - 1 degrees C/min, - 5 degrees C/min, and the recovery and viability of MNC and CD34+ cells, the clonogenic capacity and the ex vivo expansion potential of UCB progenitor cells were evaluated after thawing. With - 1 degrees C/min cooling rate, the recovery of MNC reached 93.3% +/- 1.8% , viability 95.0% +/- 3.9% , recovery of CD34+ cells 80.0% +/- 17.9% , and clonogenic recovery were 87.1% +/- 5.5%, 88.5% +/- 8.9%, 86.2% +/- 7.4% for BFU-E CFU-GM CFU-MK, respectively. After 14 days of liquid culture, no significant difference was detected in CFC expansion between fresh and cryopreserved MNC cells with - 1 degrees C/min cooling rate, but this was not the case with - 0.5 degreesC/min and - 5 degrees C/min. In conclusion, it was demonstrated that controlling the rate at - 1 degrees C/min is more suitable for cryopreservation of hematopoietic stem cells than - 0.5 degrees C/min and - 5 degrees C/min.
Cell Survival ; physiology ; Cells, Cultured ; Cryopreservation ; methods ; Erythroid Precursor Cells ; cytology ; Fetal Blood ; cytology ; Flow Cytometry ; Granulocyte-Macrophage Progenitor Cells ; cytology ; Hematopoietic Stem Cells ; cytology ; Humans

OBJECTIVETo explore the ex vivo expansion characteristics of selected CD(34)(+) cells and mononuclear cells (MNC).

METHODSCD(34)(+) cells were isolated from umbilical cord blood MNC by MiniMACS system, expanded under the same conditions as that for MNC. The effects of re-isolation and the MNC supernatant (MNC-SN) on the selected CD(34)(+) cells were investigated. And the CD(34)(-) cells of MNC were cultured ex vivo.

RESULTSIn the culture of selected CD(34)(+) cells, both the colony density and the proportion of the CD(34)(+) cells declined continuously with the culturing, although they presented a high proliferation potential. However, in the culture of the MNC, from day 0 to day 7, the colony density and the proportion of CD(34)(+) cells were increased from 412 +/- 167/10(5) cells and (1.12 +/- 0.42)% to 1 162 +/- 566/10(5) cells and (4.17 +/- 1.44)%, respectively. It was found that both the total cells and the CD(34)(+) cells restored expansion potential by re-isolating. CD(34)(-) cells of MNC had the ability to form colony and could transform to CD(34)(+) cells. MNC-SN can promote colony forming ability and lead to CD(34)(+) cells differentiation at the same time.

CONCLUSIONSIn ex vivo culture, selected CD(34)(+) cells presented a high proliferation and differentiation potentials, and the CD(34)(-) cells produced during the cultivation had inhibition effect on CD(34)(+) cells expansion. CD(34)(-) cell population from cord blood MNC contained hematopoietic stem/progenitor cells and the cytokines secreted by CD(34)(-) cells could induce CD(34)(+) cells to more mature colony-forming cells.

Antigens, CD34 ; analysis ; Cell Count ; Cell Differentiation ; immunology ; Cell Division ; immunology ; Cells, Cultured ; Colony-Forming Units Assay ; Fetal Blood ; cytology ; immunology ; Hematopoietic Stem Cells ; cytology ; Humans ; Leukocytes, Mononuclear ; cytology ; Time Factors

Comminuted fracture of distal femur is a common lower limb injury from traffic accidents, especially from motor accidents. Routine dynamic condylar screw (DCS) or 95-degree condylar plate (CP) sometimes cannot solve the bone defect in the center of alignment and contralateral diaphysis for the reason of absent screw anchor point, especially for AO C2.2-2.3 types. Many authors recommended open reduction and fixation with less invasive stabilization system (LISS) as the treatment of choice, but there are still problems in fusion and alignment. In this study, we reported our experiences with the use of bone splint technique in the high-energy commimuted fracture of distal femur with central and medial bone defect in adolescents.
Adolescent ; Adult ; Bone Plates ; Female ; Femoral Fractures ; surgery ; Fracture Fixation, Internal ; methods ; Fractures, Comminuted ; surgery ; Humans ; Male ; Splints

Objective To explore the effect of transplantation of neural stem cells(NSCs)pretreated with brain-derived neurotrophic fac-tor(BDNF)on acute ischemic stroke in mice.Methods NSCs from a newborn(one day)C57BL/6 mouse were isolated and cultured in vi-tro.A total of 150 healthy C57BL/6 mice,ten-week-old,were randomly divided into five groups,that group A accepted sham operation,and groups B,C,D and E were subjected to focal ischemia by photothrombosis.Group D was transplanted NSCs 24 hours after ischemia,while group E transplanted NSCs pretreated with BDNF and group C accepted same volume of medium.All the groups were tested with rotarod test and grip strength one day before transplantation and three,seven,14,21 and 28 days after transplantation.The differentiation of NSCs in groups D and E were observed immunofluorescence staining of microtubule-associated protein-2(MAP-2)and glial fibrillary acidic pro-tein(GFAP).Results The time on rotarod arranged from more to less was groups E,D and B(P<0.05)three,seven,14,21 and 28 days after transplantation,as well as grip strength 14,21 and 28 days after transplantation(P<0.05).The Edu/GFAP positive cells and Edu/MAP-2 pos-itive cells were found in both groups D and E.Conclusion Transplanting NSCs can promote the behavioral recovery after ischemic stroke, and it is more effective as pretreated with BDNF.

OBJECTIVETo assess the metastatic frequency of subcarinal lymph nodes of thoracic esophageal carcinoma and its influencing factors, in order to determine the adequate range of lymph node dissection during esophagectomy.

METHODSThe clinical data from a cohort of 782 patients with thoracic esophageal carcinoma who underwent esophagectomy with lymphadenectomy were analyzed retrospectively with respect to the impact of subcarinal lymph nodes dissection or no dissection on the incidence of postoperative complications.

RESULTSThe metastasis rate of subcarinal lymph nodes was 17.5%. The metastasis rates in the upper, middle and lower esophageal carcinomas were 8.3%, 19.1% and 16.2%, respectively (P > 0.05). For T1, T2, T3 and T4, they were 0, 4%, 22.2% and 34%, respectively (P < 0.05). The overall incidence of postoperative complications with and without subcarinal lymph nodes dissection was 19.0% versus 14.6% (P > 0.05), and the incidence of pulmonary complications was 10.3% versus 7.3% (P > 0.05).

CONCLUSIONSThoracic esophageal carcinomas have a high metastasis rate of subcrinal lymph nodes, and subcarinal lymph nodes dissection is not associated with increasing perioperative complications. Therefore, for the thoracic esophageal carcinoma, no matter the tumor site, esophageal cancer length or size, once the tumor invades the outer membrane, routine subcarinal lymph node dissection should be done.

Adult ; Aged ; Aged, 80 and over ; Esophageal Neoplasms ; pathology ; surgery ; Esophagectomy ; Female ; Humans ; Lymph Node Excision ; methods ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Staging ; Postoperative Complications ; Retrospective Studies