Objective To explore the relationship between serum levels of interferon-inducible protein-10 (IP-10), interferon-γ (IFN-γ) and hepatic inflammatory reaction, disease progression in patients with severe hepatitis B (SHB). Methods Sera of 40 patients with SHB at time of admission,at the beginning of single plasma exchange (PE), at time of PE completion and 5 days after PE. The SHB patients were divided into improved group and aggravated group. And 20 patients with chronic hepatitis B (CHB) and 20 healthy controls were enrolled in this study. Serum levels of IP-10, IFN-γand tumor necrosis factor-α (TNF-α) were determined by enzyme-linked immunosorbent assay (ELISA). Results The serum levels of IP-10 in patients with SHB and CHB on admission were (683.6 174.6)ng/L and (216.1 102.9)ng/L, respectively, which were notably higher than those in healthy controls [(107.6 55.8)ng/L F=9.036, both P＜0. 01],and those in patients with SHB was significantly higher than that in patients with CHB (P＜0. 01). The serum level of IFN-γ in patients with SHB and CHB on admission were (19. 8 8. 8) ng/L and (16. 7 7. 8) ng/L,respectively, which were significantly higher than those in healthy controls [(2.6 1.2) ng/L F=9. 288, both P＜0. 01]. The serum level of IP-10 and IFN-γ were both positively correlated with TNF α (r=0. 366 and r=0. 365, respectively;P＜0.05) and both negatively correlated with prothrombinase activity (r=-0.401 and r=-0.350, respectively;P＜0.05), but not correlated with serum total bilirubin(r=0. 223 and r=0. 219, respectively;P＞0.05). The serum level of IP-10 and IFN-γ were positively correlated ( r= 0. 602 ; P= 0. 000 ). On day 5 after PE, serum level of IP-10 in patients with SHB was significantly decreased compared with that'in patients before PE (t= 8. 947, P＜0.01 in improved group;t=4. 121, P＜0.05 in aggravated group) and that in aggravated group was significantly higher than improved group (t=7.862, P＜0.01). But serum level of IFN-γ was not decreased significantly (t=0. 491, P＞0.05). Conclusions IP-10 and IFN-γ are involved in the hepatic immunopathological mechanism. Serum level of IP-10 is correlated with the severity of hepatic inflammatory injury and IP-10 could reflect the progression and development of disease in patients with SHB.

Objective To observe the clinical effect of percutaneous transluminal angioplasty (PTA)combined with autologous peripheral blood stem cells(PBSC)transplantation in the treatment of lower extremity ischemic disorders.Methods Fourty-two cases of lower extremity ischemic disorders in the treatment group were treated with PTA and autologous peripheral blood stem cells injection and 40 cases in control group were treated with PTA exclusively.Results All the procedures were successful.In treatment group,ABI improved from 0.32 ±0.11 to(at the 3rd month)and 0.49 ±0.13(at the 6th month)(t=-6.765,-6.040,P＜0.05)while TcPO_2 improved from(26.1 ± 2.3)mm Hg to(32.7 ±4.2)mm Hg(at the 3rd month)and(34.5 ±2.7)mm Hg(at the 6th month)(t=-8.901,-14.250,P＜0.05).In control group,ABI improved from 0.30 ±0.12 to 0.47 ±0.15 and 0.47 ±0.130=-5.631,-5.873,P＜0.05)while TcPO_2 increased from(25.9 ±2.4)mm Hg to(28.9 ±2.9)mm Hg(at the 3rd month)and(28.9 ± 2.1)mm Hg(at the 6th month)(t=-5.090,-5.389,P＜0.05).There was significant difference in TcPO_2 on follow-up between the two groups after the treatment(P＜0.05).Conclusion Autologous PBSC transplantation in combination of PTA was effective for the treatment of lower extremity ischemic disorders.PBSC injection helps to increase TcPO_2.

Objective To detect the mutations in connexin genes in a family with hidrotic ectodermal dysplasia(HED)complicated by pseudo-ainhum.Methods Peripheral blood samples were collected from a 20-year-old patient with HED complicated by pseudo-ainhum,and from his unaffected sister.Total DNA was extracted from these samples,and PCR was performed to amplify the partial coding region of GJB2,GJB5 and GJB6 genes.Subsequently.PCR products were bidirectionally sequenced in both subjects.Results No mutation was detected in GJB5 or GJB6 gene in either subjects.Two mutations (V27I and V37I)were detected in the GJB2 gene in the patient but not in his sister.Conclusion The mutation in the GJB6 gene may be absent in patients with HED;there might be other genes involved in the pathogenesis.

Objective To explore the seed protein extraction method for Amomum villosum Lour., so as to lay the foundation for the study of differential proteomics of Amomum villosum Lour. during seed dormancy and germination. Methods We used the four kinds of commonly-used protein extraction methods for the experiment, including Tris-HCl extraction method, trichloroacetic acid (TCA) /acetone precipitation method, direct lysis buffer method and improved Tris-saturated phenol method. The obtained proteins were quantified, and then were analyzed by sodium dodecyl sulphate- polyacrylamide gel electrophoresis (SDS-PAGE) . Results The protein yield was the highest when using direct lysis buffer method, which was mainly composed of small molecule protein. The protein yield was the lowest using Tris/HCl method, characterized by the diffused stripes in SDS-PAGE spectra. The protein yield was moderate using TCA/acetone method and improved Tris-saturated phenol method, and the obtained protein by the TCA/acetone method had less stripes in SDS-PAGE spectra than that by Tris-saturated phenol method, and was characterized by macromolecular protein. Conclusion The combination of direct lysis buffer extraction method and improved Tris-saturated phenol method may provide reference for seed protein extraction method of two-dimensional electrophoresis for Amomum villosum Lour..

Objective To explore the clinical features of sentinel polyps (rectal polyps with proximal colon carcinoma), and the correlation between sentinel polyps and proximal colon carcinoma. Methods The clinical data of 331 patients with rectal polyps were retrospectively analyzed. According to the combination condition of proximal colon carcinoma, the patients were divided into sentinel polyps group (observation group, 37 cases) and pure rectal polyps group (control group, 294 cases). The family history, laboratory examination, colonoscopy, clinical pathological features, treatment, sequelae and short-term prognosis were compared between 2 groups. Results The family history rate, positive rate of tumor marker and the incidences of polyps maximum diameter>1 cm, polyps>5 pieces, adenomatous polyp in observation group were significantly higher than those in control group:35.1%(13/37) vs. 4.8%(14/294), 67.6%(25/37) vs. 6.8%(20/294), 62.2%(23/37) vs. 46.6%(137/294), 43.2%(16/37) vs. 11.9%(35/294) and 83.8%(31/37) vs. 35.4%(104/294), and there were statistical differences (P<0.01). The patients in control group did not have special find in colonoscopy. The majority patients in observation group had new organisms around the lumen growth in colonoscopy, but the intestinal canal between rectal polyps and proximal colon carcinoma did not have special find. The majority pathologic type of proximal colon carcinoma patients in observation group was papillary adenoearcinoma and tubular adenocarcinoma, 75.7%(28/37). Duke stage:A stage was in 11 cases (29.7%, 11/37), B stage in 11 cases (29.7%, 11/37), C stage in 9 cases (24.3%, 9/37), and D stage in 6 cases (16.2%, 6/37). In control group, 282 patients (95.9%, 282/294) were treated by endoscope, and they were cured and discharged. In observation group, 15 patients (40.5%, 15/37) were treated with radical operation, 9 patients (24.3%, 9/37) by endoscope, 7 patients (18.9%, 7/37) with palliative surgery, 4 patients (10.8%, 4/37) with chemotherapy, and 2 patients (5.4%, 2/37) with symptomatic treatment;the patients were followed up for 6-12 months, the 23 patients with complete tumor resection did not relapse, 12 patients showed tumor reduction or symptomatic relief, and the other 2 patients died. Conclusions If maximum diameter over 1 cm, multiple and adenomatous polyps exist, the possibility of carcinogenesis of the polyps or the proximal colon should be awared. The patients should be followed up in short-term and complete the whole colon examination.

Objective To construct the prokaryotic expression vector of mouse CD25 extracellular domain and to express it in E coli.Methods Total RNA was isolated from splenocytes of Balb/c mice.The CD25 extracellular domain gene was amplified by RT-PCR and cloned into the PET-32a vector.A positive recombinant,PET-32a-CD25e,was identified by enzyme cleaving and sequencing before its expression in E.coli,and transferred into E.coli BL21(DE3) plysS for its expression.After purification with Ni+ resin and renaturation in vitro,a relative molecular mass(Mr) of the interesting protein was detected by SDS-PAGE and Wes-tern blotting.Effect of the purified interesting protein on the proliferation of splenocytes from T cell vaccine-immunized syngeneic mice was detected by MTT assay.Results The cloned CD25 extracellular domain gene was identified to be functional by sequencing and expression.The purified interesting protein could significantly induce the proliferation and IL-4 secretion of splenocytes from T cell vaccine-immunized mice in vitro.Conclusion Mouse CD25 extracellular domain gene can be successfully expressed in prokaryotic cells with biological activity,which lays a foundation for further relative studies.

Objective:To investigate the effect of calcium ionorphore on B7 costimulatory molecules of chronic myeloid leukemia cells line K562.Methods:Well growing K562 cells were cultured in the medium containing calcium ionorphore(375 ng/ml),with K562 cells without CI treatment as control.The cells′ viability and number were calculated by Trypan Blue exclusion at 0,48 and 96 h.Before and after 96 h of cultured,B7-1 and B7-2 expression was assayed by flow cytometry.The proliferation of allogeneic human T cells was measured by MTT colorimetry.Results:B7 costimulatory molecules were abcent or lowly expressed on K562 cells.After 96 h of CI treatment,B7 costimulatory molecules of K562 cells were markedly upregulated and marked activation of allogeneic T cells occurred.No notable morphological change was found during the culture.K562 cells cultured in medium with CI grow slowly than that without CI.Conclusion:B7 costimulatory molecules expression on chronic myeloid leukemia cells line K562 surface was defective.These costimulatory molecules on K562 cells can be upregulated by calcium ionorphore.Calcium ionorphore may inhibit the growth of K562 cells.

Objective To analyse the experience and treatment of early phase severe acute pancreatitis (SAP) in intensive care units (ICU).Methods A multicenter retrospective study was done on patients with SAP treated in three major teaching hospitals (Beijing Hospital,Peking University First Hospital and Peking University Shenzhen Hospital) in China from Jan.2001 to Dec.2011.Results There were 188 patients who were enrolled in the study,including 121 males and 67 females.The age ranged from 19 to 104 (51.0±18.2) years.The mean APACHE Ⅱ score was (22.2±4.6).84.0％ of patients survived,the mortality was 10.1％ in the early phase and 5.9％ in the late phase.The most common systemic complications were acute renal injury (46.3 ％),acute respiratory distress syndrome (35.6％),and septic shock (17.6％).The local complication rate was 47.3％,which included acute peripancreatic fluid collections (32.8％),acute necrotic collection and walled-off necrosis (48.4 ％) and pseudocyst (18.8 ％).The conservative treatments included intensive care,fluid resuscitation,mechanical ventilation,continuous renal replacement therapy,antibiotics,glucose control,inhibition of pancreatic enzyme activity and secretion,and nutritional support.Surgical intervention included endoscopic retrospective cholangio-pancreatography and endoscopic sphincterectomy,B ultrasound or CT guided puncture and drainage,and surgical drainage and debridement of necrosis.Conclusions The early phase of SAP was characterized by systemic inflammatory response syndrome and multiple organ dysfunction syndrome which accounted for the first peak in mortality.Intensive care therapy and multi disciplinary comprehensive combined strategy were very important for these patients with systemic and local complications.ICU treatment in the early phase was preferred for patients with SAP.

Objective To investigate the association between tumor necrosis factor ? (TNF ?) gene polymorphism and ankylosing spondylitis (AS).Methods Genomic DNA from 98 Chinese AS patients and 70 ethnically matched controls were typed for TNF(308) polymorphism by allele specific polymerase chain reaction (AS PCR).Results The TNF genotypes in AS patients were respectively TNF1 homozygote 37%,TNF2 homozygote 10% and TNF1 and TNF2 heterozygote 53%.While TNF genotypes in controls group were respectively TNF1 homozygote 67%,TNF2 homozygote 3% and TNF1 and TNF2 heterozygote 30%.Significant difference was found in the distribution of TNF 308 genotype between both groups ( ? 2=15 73, P

Objective To investigate the effect of anti-Mlllerian hormone (AMH) on hormone secretion and P450 aromatase mRNA expression from cultured human luteinized granulosa cells. Methods Human luteinized granulose cells were derived from 10 patients treated by in vitro fertilization-embryo transplantation (IVF-ET) in the Second Affiliated Hospital of Sun Yat-sen University from June to December 2006. Granulose cells were divided into group A, B, C, D, E depending on different concentration of AMH,testosterone group and blank group. 1×10-7moL/L testosterone and 1,5,10,20,50 μg/L AMH were added into the culture medium of group A,B,C,D and E. 1×10-7mol/L testosterone was added into the culture medium of testosterone group while no other ingredient was added into the medium of blank group. Estrogen levels in supernates were measured at 24,48,72 hours after cell incubation. RT-PCR was performed to detect the P450 aromatase mRNA expression in group B, C, D, E and testosterone group at 72 hours after cell incubation. Results (1) Estrogen levels in supernates of granulose cell culture at 24,48,72 hours were (8.529±0.381)×104, (10.977±0.436)×104, (13.309±0.506)×104 pmol/L in group A, (7.027±0.276)×104, (9.167±0.300)×104, (10.794±0.555)×104 pmol/L in group B, (6.039±0.226)×104,(7.585±0.548)×104, (8.797±0.518)×104 pmol/L in group C, (5.118±0.460)×104, (5.716±0.496)×104, (6.205±0.667)×104 pmol/L in group D, (4.932±0.148)×104, (5.323±0.184)×104,(5.629±0.212)×104 pmol/L in group E. When compared with blank group [(0.001±0.001)×104,(0.006±0.003)×104, (0.029±0.011)×104 pmol/L], the statistical differences were observed in group A,B,C,D,E(P<0.01) ; when compared with testosterone group [ (8.418±0.569)×104, (10.841±0.689)×104, (13.301±0.637)×104 pmol/L], the statistical differences were observed in group B,C,D and E(P<0.01) ; statistical differences were also observed in group C, D and E when compared with group B, and also group D and E when compared with group C(P<0.01). No significant difference was observed between group D and E (P>0.05). In group A, B, C, D, E and testosterone group, the estrogen levels at 24 hours after cell culture were significantly lower than those at 48 and 72 hours (P<0.01) ; statistical difference was observed between estrogen levels at 48 and 72 hours(P<0.01). No significant difference was observed among 24,48 and 72 hours in blank group (P>0.05). (2) Relative ratios of intensity of P450 aromatase/β-actin at72 hours of cell culture in group B,C,D and E were 0.6148±0.0046, 0.5156±0.0012, 0.4698±0.0027 and 0.4282±0.0017, respectively, which were statistically lower than that in testosterone group (0.8224±0.0021, P<0.01) ;statistical differences were also observed in group C, D and E when compared with group B, and also group D and E when compared with group C(P<0.01). No significant difference was observed between group D and E (P>0.05). Conclusion It is suggested that AMH might affect estrogen synthesis by inhibiting P450 aromatose activity so that lead to hyperandrogenism microenvironment in local ovary.