Objective To investigate the distribution and antimicrobial resistance of clinically isolated pathogens from geriatrics department of a hospital in 2013,so as to provide reference for clinical antimicrobial use.Methods Distribution and antimicrobial resistance of pathogens isolated from patients in a geriatrics department between Janu-ary and December 2013 were analyzed statistically.Results Of 1 896 pathogenic strains,1 289(67.99%)were gram-negative bacteria,439 (26.00%)were gram-positive bacteria,and 114(6.01%)were fungi;the top 4 isolated pathogens were Klebsiella pneumoniae ,Pseudomonas aeruginosa ,Acinetobacter baumannii ,and Escherichia coli .Extended-spec-trumβ-lactamase (ESBL)-producing Escherichia coli and Klebsiella pneumoniae accounted for 53.26% and 31.10% of Escherichia coli and Klebsiella pneumoniae respectively;Of Staphylococcus aureus and Staphylococcus epidermidis strains,methicillin-resistant isolates (MRSA and MRSE)accounted for 22.47% and 80.00%,respectively,of Enterococ-cus strains,vancomycin-resistant isolates (VRE)was 3.10%,Klebsiella pneumoniae and Enterobacteriaceae were highly sensitive to imipenem,meropenem and ertapenem.Resistant rate of Acinetobacter baumannii to imipenem and meropenem was 79.48% and 80.35% respectively,Pseudomonas aeruginosa had the lowest resistant rate to amikacin (10.70%). Gram-positive bacteria were highly sensitive to vancomycin and linezolid.Conclusion The major pathogens isolated from the elderly patients are gram-negative bacteria,and antimicrobial resistance is serious,surveillance of antimicrobial resist-ance is important for rational use of antimicrobial agents and control of pathogen resistance.

Objective To study the clinical significance of a novel marker of platelet clumps count provided by hematology analyzer in differentiating true thrombocytopenia from EDTA-dependent pseudothrombocytopenia (EDTA-PTCP). Methods Samples from 65 cases of thrombocytopenia (including 15 EDTA-PCTP samples and 50 random samples of true thrombocytopenia) and 50 healthy controls were analyzed using hematology analyzers, and samples with low platelet counts were checked by replacing citric acid and using manual microscope observation to identify true thrombocytopenia from EDTA-PTCP. A novel marker of platelet clumps count was used to differentiate the two diseases for samples anficoagulated with EDTA or citric acid. Results In 65 patients with thrombocytopenia, platelet counts were (48±11)×109/L detected by automatic hematology analyzers. Fifty of 65 cases were true thrombocytopenia which showed low platelet counts [(48±10)×109/L by automated analyzer and (46±11)×109/L by manual assay]. No significance was observed between them (t=-1.26, P0.05). Platelet clumps counts were 86±15. No platelet clamps were detected under microscope. The other 15 cases were EDTA-PTCP [platelet counts were (48±12)×109/L and platelet clumps counts (840±184) were increased significantly by automated analyzer and using EDTA anticoagulant] which showed obviously platelet clumps and no less platelet counts under microscope. After replacing citric acid, platelet counts [(141±13)×109/L by automated analyzer and (134±17)×109/L by manual microscope assay] were increased significantly. No significance was observed between them (t=-1.29, P0.05). Platelet clumps counts (75±12) were decreased obviously compared with EDTA anticoagulant method (t=-6.82, P<0.001). No platelet clumps were detected under microscope. Conclusion Platelet clumps counts may be a useful clinical indicator for monitoring of platelet aggregates, especially for EDTA-PTCP caused by platelet clumping.

Objective To evaluate the efficacy of pegylated interferon α-2a and entecavir (ETV) combination therapy for patients with HBeAg positive chronic hepatitis B (CHB).Methods Fifty eight HBeAg positive CHB patients were assigned to two groups:29 patients received ETV 0.5 mg daily for 72 weeks (ETV group) and 29 patients received ETV and pegylated interferon α-2a 180 μg weekly for 48 weeks followed by ETV alone for 24 weeks (combination group).Serum samples were collected from all patients every 12 weeks for assessment of biochemical,virological and serological responses to treatment.Results Fifty four patients completed the 72-week study,including 28 in ETV group and 26 in combination group.There were no significant differences in week 24,week 48 and week 72 of ALT normalization [72％ (21/29)vs.93％ (27/29),x2 =2.104;90％ (26/29) vs.97％ (28/29),x2 =0.269;90％ (26/29) vs.97％ (28/29),x2 =0.269],HBV DNA undetectable rate [31％ (8/26) vs.46％ (13/28),x2 =1.391;62％ (16/26) vs.57％ (16/28),x2 =0.108;77％ (20/26) vs.75％ (21/28),x2 =0.027],HBeAg loss rate[12％(3/26) vs.25％ (7/28),x2 =0.850;31％ (8/26) vs.32％ (9/28),x2 =0.012;46％ (12/26) vs.36％(10/28),x2 =0.609] and HBsAg levels (log10 IU/ml) (3.63 ± 0.45 vs.3.36 ± 1.18,t =-1.066;3.45 ±0.43 vs.3.23 ± 1.15,t =-0.915;3.36 ± 0.58 vs.2.88 ± 1.28,t =-1.762) between two regimens (all P ＞ 0.05).Among 58 patients,15 were HBeAg and anti-HBe double-positive (26％)and 43 were HBeAg mono-positive patients.The baseline HBV DNA level [(5.07 ± 1.50) vs.(6.40 ± 1.47) log10 IU/ml,t =2.858,P ＜ 0.05] and HBeAg titer [14 (4-45) vs.732 (296-1 012) S/CO,Z =-5.031,P =0.05] in double-positive patients were lower than those in mono-positive patients.The HBV DNA undetectable rate of double-positive patients was significantly higher than that of mono-positive patients in 24 weeks [10/15 vs.26％ (10/39),x2 =7.819,P ＜0.05] and 72 weeks [15/15 vs.69％ (27/39),x2 =4.287,P =0.05].The HBeAg loss rate of double-positive patients was higher than that of mono-positive patients in 12 weeks [6/15 vs.10％ (4/39),x2 =4.533,P =0.05] and 48 weeks [9/15 vs.26％ (10/39),x2 =5.608,P =0.018].This tendency was more significant in the combination therapy group,but the difference was not statistically significant.(5/6 vs.4/9,P =0.065).Conclusions Compared with Entecavir monotherapy,entecavir combined with interferon may not improve the therapeutic effect in HBeAg positive chronic hepatitis B patients.However,the therapeutic response of HBeAg/anti-HBe double-positive patients may better than that of HBeAg mono-positive patients.

Despite different opinions on its definition and classification in the past,a consensus has gradually been reached regarding the na-ming,classification,and clinical diagnosis of liver failure.The classification of liver failure is described,and the definition and natural his-tory of severe exacerbation of hepatitis B are summarized.Antiviral treatment and artificial liver support in the early stage are beneficial for clinical outcomes and prognosis.

Background Researches showed that CD4+CD25+ natural regulatory T cells (nTregs) play an important role in maintaining peripheral immune tolerance, while immunotherapy using in vitro-expanded induced regulatory T cells (iTregs) suppresses allograft rejection in multiple organ transplantation.The inducing method of iTregs still needs to be optimized.Furthermore,the effect of iTregs on grafts of keratoplasty is unclear.Objective This study was to investigate the inducing and expansion method of iTregs and explore its inhibitory effects on corneal allograft rejection.Methods Bone marrow-derived dendritic cells (BMDCs) were isolated from C57BL/6 mice femora and cultured.CD4+ CD25+ T cells and CD4+ CD25-T cells were isolated from mouse spleen and separated using flow cytometry.The CD4+CD25-T cells were divided into negative control group (CD4+CD25-T cells), CD3/ 28 antibody bead group (CD4+CD25-T cells+CD3/28 antibody bead) ,2.5 ng/ml transforming growth factor (TGF)-β1 induced group and 10.0 ng/ml TGF-β1 induced group.The iTregs was formed after induction of different concentrations of TGF-β1 and CD3/CD28 antibody bead (1 : 1).CD3/CD28 antibody bead (1 : 2) , interleukin-2 (IL-2) and TGF-β1 were used to expand iTregs.The phenotype and proliferation of iTregs were assayed by flow cytometry,and the inhibitory effect of iTregs on effector T cells (Teffs) was analyzed by mixed lymphocyte reaction.Allogenic keratoplasty model (C57BL/6→BALB/c) was build,and 0.1 ml iTregs or nTregs suspension or PBS was injected via posterior venous plexus of fellow eyes to assess the graft survival time.The use and care of the mice followed the ARVO statement.Results The CD4+CD25+ T cell proportions were (6±3)％ ,(91±4)％ ,(91±3)％ and (86± 6) ％ in the negative control group,CD3/CD28 antibody bead group, 2.5 ng/ml TGF-β1 induced group and 10.0 ng/ml TGF-β1induced group, showing significant increases in the CD3/CD28 antibody bead group, 2.5 ng/ml TGF-β1 induced group and 10.0 ng/ml TGF-β1 induced group compared with the negative control group (all at P＜0.01).The Foxp3+ T cell proportions of the CD3/CD28 antibody bead group,2.5 ng/ml TGF-β1 induced group and 10.0 ng/ml TGF-β1 induced group were (1.18 ±0.20) ％ , (8.70± 1.80) ％ and (21.80±3.36) ％ , showing significant increases in the 2.5 ng/ml TGF-β1 induced group and 10.0 ng/ml TGF-β1 induced group compared with the CD3/CD28 antibody bead group (both at P＜0.01).Compared with the nTregs, the expression of CD69 was lower, and the expressions of PD-1 and Foxp3 were raised in the iTregs (all at P＜0.01).The proliferation of Teffs were decreased when cocultured with iTregs in comparison with nTregs at 1 : 1,1 : 2,1 : 4,1 : 8,1 : 16 Tregs/Teffs rations (all at P＜ 0.01).The survival time of mouse corneal grafts was 4 weeks with the permanent tolerance of 50％ in the iTregs injected group,which was superior to the 3 weeks survival time and 17％ permanent tolerance in the nTregs injected group(P＜0.05).Conclusions TGF-β1 can induce CD4+ CD25-T cells to form iTregs, which highly express Foxp3.iTregs show a stronger inhibitory effect on the growth of lymphocytes than nTregs, and therefore suppress the graft rejection after keratoplasty.

[Objective] To discuss the surgical treatment strategy of spinal hemangioma.[Methods]Data on 33 patients with spinal hemangioma treated from October 1997 to October 2008 were reviewed.There were 19 females and 14 males.The average age was 54 years old.Chief complaints were pain(n=13),myelopathy(n=12),radiculopathy plus local pain(n=8).Nine patients without spinal cord and radial nerve compression underwent percutaneous vertebroplasty(PVP),and 24 patients with spinal cord and radial nerve compression,paravertebral soft tissue mass or spinal unstability were treated by operation.Anterior,anteroposterior and posterior approaches were used in 9,12,and 3 patients respectively.Transarterial embolization was done in 3 patients.[Results]No perioperative complication was observed in 9 patients after PVP,with a mean follow-up of 41 months.Pain was relieved.There was no perioperative death among the 24 patients who received operation.The blood loss during operation was 2739 ml and 1619 ml for anterior and posterior approaches,respectively(P=0.12).One case of paravertebral hematoma,one case of spinal canal hematoma,and one case of wound infection were observed in this group.Pain was relieved in 89%(17/19)of the patients.According to the Frankel Scale,the neurologic situation was improved in all the 12 patients with myelopathy.By a mean follow-up of 48 months,no internal fixation failure was seen.Tumor recurrence or growth was found in 4 patients,two had no symptoms and one underwent radiotherapy for pain.Repeated operation was performed in one patient with myelopathy.[Conclusion]Most patients with symptomatic spinal hemangioma could be treated successfully by surgery.Individual surgical plan should be made according to the reason causing symptoms and the general condition of the patient.

[Objective]To retrospectively study all the low grade central osteosarcoma(LGCO) cases treated in our center in about a past decade.[Method]Thirteen cases with LGCO treated from January 1998 to June 2008 were studied.The mean age at diagnosis was 31-year-old.The anatomical location of the lesions was similar to that of a conventional osteosarcoma.Pain and mass were common complaints with a mean duration of 16 months before diagnosis.Eight cases accepted the first operation in our center,while the other 5 cases had undergone operations in other hospitals with an intralesional resection in four and a marginal resection in one.Eleven patients had an Enneking stage I disease and two had a stage III disease.The first choice for patients with stage I disease was wide resection.[Result]In two patients with local recurrence,the postoperative pathological diagnoses were fibrosarcoma and malignant fibrous histocytoma respectively.The metastatic lesions were found to be a LGCO in both patients with stage III disease.The average follow up duration was 29 months.Lung metastasis occurred in one stage I patient 15 months after operation.Gama-knife was used to get rid of the lung lesions and then systemic chemotherapy was given.Now he was still alive with disease 27 months after the first operation.One of the 2 patients with stage III disease died of the disease 26 months after operation and the other one was failed to follow up 6 months after operation.The other 10 patients were alive without disease at the last follow-up.For those who received limb salvage operations,the mean MSTS 93 score was 79%.[Conclusion]For most patients with LGCO,wide resection of tumor is preferred.Good oncologic and functional results could be expected after a limb salvage operation without chemotherapy.Insufficient resection of tumor would lead to recurrence.Long time follow up is compulsory.High grade sarcomas would be found in some of the local recurrent lesions.Metastasis would occur in few patients and the metastatic lesions may still be a LGCO.

Objective To discuss the techniques of tumor resection, pelvic reconstruction and postoperative management by reviewing the patients with malignant pelvic tumors in recent years. Methods 76 patients with primary malignant pelvic tumors were treated operatively between July 1997 and July 2003. The series comprised 47 males and 29 females. 31 cases were diagnosed as chondrosarcoma, 15 as Ewing sarcoma, 7 as osteosarcoma, 3 as lymphoma, 3 as malignant fibrohistiocytoma (MFH), 1 as hemangiopericytoma, 2 as myeloma, 13 as giant cell tumor(GCT). According to Enneking's division, the most common region of the primary pelvic tumor involved was region Ⅱ (51 cases), followed by region Ⅰ (16 cases) and region Ⅲ (9 cases). 16 cases were reconstructed with threaded pins and cement after resection of the ilium. 9 patients had local resection of pubis and ischium. 21 patients had hemipelvectomy. Among 30 patients with periacetabular tumors, 8 were reconstructed with hemipelvic prosthesis, 7 with saddle prosthesis, 6 with replantation of cauterized tumor bone and total hip arthroplasty, 9 with cemented total hip replacement after curettage of lesion. Results After 16 months to 6 years follow-up, among the 21 patients with hemipelvectomy, 4 patients had local relapse because the sacrum invaded by the tumors. 7 of 25 patients with lesions in regionⅠ or region Ⅲ had local relapse postoperatively due to the marginal incision to the acetabula. Among the 21 patients who had tumor resection and reconstruction in region Ⅱ, 4 had local relapse, 3 of which reconstructed with cauterized hemipelvis. As for functional recovery, all of the 25 patients with lesions in region Ⅰand region Ⅲcould walk postoperatively without assistance of a cane. 21 patients with acetabular reconstruction after resection of lesions in region Ⅱ could sit and stand normally and walked with a cane, some of them even had normal gait. Conclusion The major principle of treatment for primary pelvic tumor is that en bloc resection, only then the functional reconstruction of pelvic ring can be considered. Operators should be acquainted with advantages and shortcomings of different reconstructive methods of acetabulum to prevent the complications.

Objective To discuss the treatment and prognosis of multifocal osteosarcoma. Methods Between January 1998 and June 2005, nine patients(seven males and two females), six with synchronous multifocal osteosarcoma and three with metachronous multifocal osteosarcoma were treated in our department. The diagnosis of multifocal osteosarcoma was confirmed by clinical presentation, radiology and pathology. Eight of the nine patients received chemotherapy with adriamycin(60 mg/m2), cisplatinum(100 mg/m2), high-dose methotrexate(8-12 g/m2), vincristine(1.4 mg/m2), ifosfamide(12.5 g/m2), and G-CSF. Based on the response to primary chemotherapy, further chemotherapy plans were made, and different operations were done to control local lesions or to relieve symptoms. Arsenous acid, paclitaxel and VP-16 were used in patients who responded poorly to primary chemotherapy. Results One patient who given up remedy was died in systematic fail due to the cancer after 5 month, five patients who were diagnosed synchronous multifocal osteosarcoma died from progressive disease in 3.5 to 17 months (mean time was 9.8 months). Only two of them had a good response to chemotherapy and had finished at least four cycles of chemotherapy,and they had a better survival time of 15 and 17 months respectively. Three patients with metachronous multifocal osteosarcoma had a better prognosis. After a two-year follow-up, two lived with disease and one lived without disease. Conclusion The prognosis of patients with multifocal osteosarcoma is poor, and patients with metachronous multifocal osteosarcoma have a relatively better prognosis. To improve the prognosis of patients with multifocal osteosarcoma, aggressive chemother-apeutics are needed.

Objective To investigate the crosstalk between Wnt signaling pathway and TGF-? signaling pathway in prostate cancer cell line PC-3. Methods Luciferase report assay and RT-PCR methods were used to explore the crosstalk between two pathways. Results Wnt signaling pathway activated TGF-? responsive CAGA-luciferase reporter activity. ?-catenin(WT), TCF-4(WT), GSK3? (KM) activated while TCF-4(DN) inhibited CAGA reporter activity. ?-catenin(WT), TCF-4(WT) synergized to activate TGF-? signaling while TCF-4(DN) had converse effect.TGF-? signaling pathway activated LEF-1 luciferase activity through a Smad3-dependent manner.Smad7 activated LEF-1 luciferase activity,Smad3 and Smad4 also synergically activated LEF-1 reporter activity.?-catenin(WT),TCF-4(WT) cooperated with each other to the activity of cyclinD1 promoter luciferase while Smad3 inhibited cyclinD1 promoter activity.The inhibitory effect of Smad3 on cyclinD1 promoter could be partially reversed by co-transfecting ?-catenin(WT),TCF-4(WT).TGF-? induced the expression of VEGF in PC-3 cells and this effect was enchanced by LiCl addition. Conclusions Wnt signaling pathway and TGF-? signaling pathway can activate each other in prostate cancer cell line PC-3.Two pathways have crosstalk and this might be important for the development and progression of prostate carcinoma.