Objective To apply fluo-rescencc-activated cell sorting(FACS) in sorting T lymphocyte (CD_3~+) and granulocyte (CD_(15)~+), which establish the separating method of series of cells from human peripheral blood, so that the scientific research and clinical research could be carried out were specifically. Methods 10 cases of normal peripheral blood were collected and T lymphocyte (CD_3~+) and granulocyte (CD_(15)~+) were stained with florescence conjugated antibodies. The positive cells were sorted by FACS. Results Before sorting the peripheral blood, the proportion of the T lymphocyte(CD_3~+) and granulocyte (CD_(15)~+) in the leukocyte is 48.8 % and 30.8 %; after sorting by FACS, the purity of T lymphocytes (CD_3~+) is up to 98 % and the recovery is about 95 %; the purity of granulocyte (CD_(15)~+) is up to 97 % and the recovery is about 96 %. Conclusion The FACS could Mlow us to quickly sort T lymphocyte (CD_3~+) and granulocyte (CD_(15)~+) with higher recovery and higher purity from the peripheral blood.

Carcinoma in Situ ; diagnosis ; pathology ; surgery ; Carcinoma, Transitional Cell ; diagnosis ; pathology ; surgery ; Cystectomy ; Humans ; Neoplasm Invasiveness ; Neoplasm Staging ; methods ; Urinary Bladder ; pathology ; surgery ; Urinary Bladder Neoplasms ; diagnosis ; pathology ; surgery

Adult ; Calcinosis ; Heart Aneurysm ; pathology ; Heart Ventricles ; Humans ; Male ; Thrombosis ; pathology

Objective To explore the enhancement effects and mechanisms of IL-32β on human cervical carcinoma cells C33A to hypoxic/hypoglycemic condition.Methods After cultured in hypoxia/hypoglycemic circumstance and normal circumstance for 20 hours respectively, the mRNA and protein expression of IL-32β in C33A cells were detected by real time-polymerase chain reaction (RT-PCR) and Western blotting respectively.Trypan blue stain was used to detect C33A cells viability in hypoxia/hypoglycemic circumstance and adding 10, 100,500 ng/ml IL-32β circumstance.The xenografted tumor of nude mice was established by intraperitoneal injection, and their volumes were tested for a given time after injecting 0, 1.0 mg/kg IL-32β.siRNA was used to construct IL-32β knockdown cells and detect the expression of VEGF.Results Under the hypoxia/hypoglycemic circumstance, the expressions of IL-32β mRNA were (6.12 ± 0.03) times of the normal circumstance (F =43.16, P ＜ 0.05), the expressions of IL-32β protein were (2.23 ± 0.04) times of the normal circumstance (F =22.32, P ＜ 0.05).The C33A cells viability in hypoxia/hypoglycemic circumstance was (51.92 ± 3.41) ％, whereas, viability in 10 ng/ml IL-32β group was (55.23 ± 3.92) ％ (F =14.25, P ＜ 0.05), viability in 100 ng/ml IL-32β group was (62.52 ± 4.14) ％ (F =35.53, P ＜ 0.01), viability in 500 ng/ml IL-32β group was (69.14 ± 2.45) ％ (F =56.28, P ＜ 0.01).After 28 days, the volume of xenografted tumor of 0 mg/kg IL-32β group was (578 ± 64)mm3, and 1.0 mg/kg IL-32β group up to (1 402 ± 142) mm3 (F =27.84, P ＜ 0.01).In addition, compared with control group, the expression of VEGF in IL-32β knockdown C33A cells was significantly decreased (F =36.85, P ＜ 0.05).Conclusion IL-32β can enhance the resistance to hypoxic/hypoglycemic condition of C33A cells, which is associated with the increase of VEGF.

Objective To explore the relationship between CD+4CDhigh25CDlow127 Treg level of peripheral blood and NHL.Methods The level of peripheral blood CD+4CDhigh25CDlow127 Treg of 123 newly diagnosed NHL and 41 healthy adults were test by flow cytometry.Results The average peripheral blood CD+4CDhigh25CDlow127 Treg level were 5.0 ％ (2.7 ％) and 7.2 ％(3.8 ％) in healthy adults and newly diagnosed NHL patients,respectively.The difference between the two groups was statistically significant(Z=-2.816 P=0.001 ).The peripheral blood Treg level was significantly higher in the male NHL patients than that in the female (7.4％ vs 6.1％,Z=- 2.729,P=0.006).The peripheral blood Treg level was significantly higher in elevated LDH patients than that in normal LDH patients(7.8％ vs 7.0％,Z=-2.623 P=0.008).The peripheral blood Treg level was significantly higher in stage Ⅲ-Ⅳ patients than that in stage Ⅰ -Ⅱ patients (7.3％ vs 5.9％,Z=-2.057 P=0.04). There was no significant relationship between peripheral blood Treg level and age, IPI score,extranodal involvement,performance status,bulky disease,pathologic subtype or B symptom (P ＞0.05).Conclusions NHL patients are in an immunosuppresive statue.Patients of male,with inreased LDH level or in stage Ⅲ-Ⅳ have higher peripheral blood CD+4CDhigh25CDlow127 Treg level.

Objective To detect peripheral blood level of CD4+ CD25high CD127low regulatory T (Treg) cells in patients with B cell non-Hodgkin lymphoma (B-NHL) and to explore its clinical significance.Methods The levels of peripheral blood CD4+ CD25high CD127low Treg cells of 100 newly diagnosed patients with B-NHL and 50 healthy adults were detected and analyzed by flow cytometry.Results The levels of peripheral blood CD4+ CD25high CD12low Treg cells were 5.00 ％ and 7.20 ％ in healthy adults and newly diagnosed B-NHL patients,respectively.The difference was statistically significant (P ＜ 0.001).The level of peripheral blood CD4+ CD25high CD127low Treg cells was significantly higher in the male patients than that in the female (P ＜ 0.01).The level of peripheral blood CD4+ CD25high CD127low Treg cells was significantly higher in elevated LDH patients than that in normal LDH patients (P ＜ 0.01).The level of peripheral blood CD4+ CD25high CD127low Treg cells in stage Ⅲ-Ⅳ patients was significantly higher than that in stage Ⅰ-Ⅱ patients (P ＜ 0.01).The level of peripheral blood CD4+ CD25high CD127low Treg cells in B symptom patients was significantly higher than that in no B symptom patients (P ＜ 0.01).There was no significant relationship between the level of peripheral blood Treg cells and age,IPI score,bulky disease or pathologic subtype (all P ＞ 0.05).Conclusions B-NHL patients were in an immunosuppresive statue.Male,elevated LDH level,B symptom and Ⅲ-Ⅳ stage were correlated with higher level of peripheral blood CD4+ CD25high CD127low Treg cells.Level of peripheral blood Treg cells may be a useful prognostic factor for B-NHL.

0.05),but there was a significant difference in the lowest SaO2,average SaO2 and AHI between the 2 groups(P

One hundred and seventy-two strains of endophytic fungi were isolated from Taxus mairei,Cephalotaxus fortunei and Torreya grandis cv.merrillia.The result of the antifungal assay shows that ninety strains of the fungi have antagonism against one or more botanical pathogenic fungi,such as Neurospora sp.,Trichoderma sp.,Fusarium sp.etc.The percentage of antifungal strains to tested strains are as follows:40% Cephalotaxus fortunei,54.2% Taxus mairei,57.1% Torreya grandis cv.merrillia.Thirty-five strains have high antifungal activities,and their inhibition zone diameter is at least 15mm.The active endophytic fungi were identified as 18 genera,most of which belong to Paecilomyces and Fusarium etc.

Mycoepoxydiene is a novel antitumor agent extracted from marine lignicolous fungi HLY-2, which is Diaporthe phaseolorum by molecule identification. The medium optimization for mycoepoxydiene by orthogonal design and the comparison of submerged fermentation and solid state fermentation were studied. The rusult is that the maximal yield of the compound is 543mg/L, which is 43 times compared to the customary half-seawater PD medium and 15 times to the best submerged condition. This optimum culture medium included potato 250g/L, seawater 300mL/L, glucose 30g/L, lactose 50g/L, KH_ 2 PO_ 4 0.65mmol/L and (NH_ 4 )_ 2 SO_ 4 1g/L in the solid state condition. Differentiation analysis between submerged and solid state fermentation, and antitumor activity of these ferment products were also studied. The antitumor activity of products of the optimum medium approached the pure compound.

AIM:To synthesize the human platelet factor-4(hPF4) gene with a convenient and effective approach, and high express the hPF4 gene in E. coli BL21 (DE3). METHODS: According to the primary structure of hPF4, the nucleotide sequence was synthesized using touch-down PCR method. The resultant gene fragment containing EcoR Ⅰ and Xho Ⅰ overhangs at 5' and 3' ends was cloned into the expression vector pGEX-4T-1 to construct the recombinant plasmid pGEX-4T-1-hPF4,which was then transformed into the E. coli strain BL21 (DE3). RESULTS: hPF4 gene was successfully synthesized by touchdown PCR method. A fusion protein composed of glutathione S-transferase (GST) and the recombinant hPF4 was expressed in BL21(DE3) by IPTG induction. The expression level of the fusion protein in E. coli was about 30% of the total cellular protein. CONCLUSION: Touch-down PCR may provide a convenient and effective approach to obtain other target genes. The expressed fusion protein forms the inclusion bodies, providing sufficient material for further purification and biological activities process.