An recombinant vector pCSV-EPO for expression of EPO cDNA in mammalian cells was constructed by the techniques of gene recombinant, PCR amplification and region-specific mutagenesis. The pCSV-EPO was introduced into COS7 cells by DEAE-dextran-mediated transfection. The expression of the EPO was demonstrated by EPO-ELISA assay. At 48h post transfection, the EPO level was 25ng/ml and 72 h was 17ng/ml.

BACKGROUND: In patients with obstructive sleep apnea(OSA),the degree of severity of OSA is mostly related to the sleep time spent in the supine position. According to the apnea-hypopnea index(AHI), positional and n onpositional OSA has been distinguished.OBJECTIVE: To compare the clinical characteristics of positional and nonpositional OSA and investigatethe relationship between sleep position and respiratory function of patients with OSA.DESIGN: A comparative clinica] observation based on the OSA patients.SETTING: Center of Sleep Disorders,Nanjing General Hospital of Nanjing Military Area Command of Chinese PLA.PARTICIPANTS: From October 1998 to May 2002,225 patients were examined with overnight polysomnography in the Center of Sleep Disorders,Department of Respiratory Disease, Nanjing General Hospital of Nanjing Command. According to the inclusion criteria of ① AHI≥10 times/hour;② Sleep time in supine and lateral positions≥30 minutes; ③ Over 20years of age; 225 patients were selected from the total of 415 consecutive patients,including 63 with positional and 162 with nonpositional OSA.METHODS: All patients received overnight monitoring with HMS-5500polysomnography system(Respironics Inc USA) in the sleep laboratory. Queries for a detailed case history and physical examination were performed with the pulmonary functions evaluated with body p lethysmography. The clinical data and results of polysomnographic and pulmonary function tests were compared between the two groups of patients and a multiple stepwise regression analysis was employed to evaluate the determinant factors for sleep position dependence of the patients' condition.MAIN OUTCOME MEASURES: Main outcome: Comparisons of the clinical data,polysomnographic result and pulmonary function between the two groups. Secondary outcome: Correlation between classification of OSA and the clinical data.RESULTS: Totally 63 (28%) of the 225 OSA patients were identified as the positional type,who had significantly greater body mass index (BMI)then the nonpositional type patients [(43.46 ± 16.43) kg/m2 vs (23.90 ± 12.36) kg/m2,t =3. 977,P<0.01]. Except for sleeping time in lateral and supine positions, minimal SatO2 and basic SatO2 before the matching for BMI,all the other factors in polysomnograms of the two groups were significant different(P<0.05 or P<0.01). After BMI matching,all the measurements were identical to those recorded for the primary grouping with the exception of nocturnal limb movement frequency,which differed but insignificant between the two groups. The pulmonary function in the two groups was no significant difference(P>0.05) . Multiple stepwise regression analysis indicated that AHI and BMI were the two determinant factors for positional dependence of OSA with a predictive power of 26. 2%. Linear correlation analysis showed correlation of AHI and SatO2 with BMI in the whole group and nonpositional group(P<0.05 or P<0.01),but not in the positional group (P>0.05).CONCLUSION: About one-third of the OSA patients shows sleep position dependence,who may respond favorably to sleep position training. Sufficient patency can be retained in the upper airway of these patients to relieve respiratory obstruction during sleep and improve the respiratory function.

In order to study the regularity of shedding virus from infected SPF chickens and the formation of aerosol of H9N2 subtype AIV, SPF chickens were bred in a positive and negative pressure isolator. Aerosol samples were collected by AGI-30 (All Glass Impinger-30) extractor, and simultaneously trachea and cloaca samples were collected by tracheal swabs and cloacal swabs in different periods after challenged with vi- ruses. The above-mentioned samples were detected by HI, Dot-ELISA and RT-PCR methods. The results in- dicated that aerosols were isolated from the 4 days to the 43 days after inoculation. It was proved that H9N2 subtype AIV could copy themselves in respiratory passage and cloaca, and then could formation of aerosols. AIV H9N2 subtype could be isolated from cloacal and tracheal swabs 3 days after inoculation and lasted for 45 days, viruses were detected from all infected SPF chickens on 7 days.

BACKGROUNDThe induction of immune tolerance and suppression of allograft rejection has become the focus in the study of liver transplantation. The effect of immune therapy with anti-CD40L mAb alone or in combination with cyclosporine A (CsA) on the recipient survival and Th1/Th2 cytokine profile was studied to elucidate its immunological mechanism and role in rat orthotopic liver transplantation.

METHODSThe model of rat orthotopic liver transplantation was established by modified Kamada's technique. Recipients were divided into group A (control group): SD-->SD; group B (group of rejection): SD-->Wistar without any treatment; group C: SD-->Wistar with CsA monotherapy from day 1 to day 5; and group D: SD-->Wistar with CsA from day 1 to day 5 and anti-CD40L mAb on day 0 and day 2. The survival of the recipients in all groups was observed and ELISA technique was used to detect the level of cytokines in peripheral blood on post-transplant day 7.

RESULTSThe survival period of recipients in groups A (> 60 days) and D (> 60 days) was significantly longer than that in group B (13.8 +/- 2.4 days). The serum levels of interleukin 2 (IL-2) and interferon gamma in group B were significantly higher than those in other groups; the level of tumor necrosis factor alpha was higher but not statistically significant. In contrast, the serum levels of IL-4 and IL-10 in group D were elevated more significantly than those in group B (P < 0.05).

CONCLUSIONSCombined immune therapy can prolong the survival of allografts. Increased expression of Th2 cytokines, which is closely related to the induction of tolerance and suppression of rejection, is beneficial to the long-term survival of recipients and allografts.

Animals ; Antibodies, Monoclonal ; therapeutic use ; CD40 Ligand ; antagonists & inhibitors ; Cyclosporine ; therapeutic use ; Cytokines ; blood ; Enzyme-Linked Immunosorbent Assay ; Graft Survival ; Immunosuppressive Agents ; therapeutic use ; Liver Transplantation ; Male ; Rats ; Rats, Sprague-Dawley ; Th1 Cells ; immunology ; Th2 Cells ; immunology

OBJECTIVETo investigate the effect of c-kit mutation on the prognosis of gastrointestinal stromal tumors.

METHODSA search of studies in PubMed and MedLine (from 1999 to 2008) was performed to assess the effect of c-kit mutation on the prognosis of gastrointestinal stromal tumors. The articles were retrieved with the entries of "gastrointestinal stromal tumors", "imatinib", "c-kit" and "mutation". A meta-analysis was performed to assess the data included.

RESULTSA total of 15 articles were collected in this analysis. No significant differences was found in incidence of mitoses (> 5/50 HPF) between the patients with wild type c-kit (wild type group) and the ones with mutated c-kit (mutation group) (P = 0.710); tumor recurrence and metastasis rate after surgery was significant higher in the mutation group than that in wild type group (P = 0.010); as for imatinib response with different c-kit mutation types, the results showed the incidence of clinical response (complete response + partial response) was significantly higher in mutation group than that in wild type group (P = 0.009), but the imatinib resistance rate was lower in mutation group (P = 0.000); three studies provided data for imatinib resistance with c-kit second mutations, the results showed the second mutations mainly focus on exon 13, 14, 17.

CONCLUSIONSC-kit mutation is related closely with the incidence of recurrence and metastasis in GIST after surgery. The mutations of c-kit influences the therapeutic effects of imatinib.

Antineoplastic Agents ; therapeutic use ; Benzamides ; Case-Control Studies ; Gastrointestinal Stromal Tumors ; drug therapy ; genetics ; Humans ; Imatinib Mesylate ; Mutation ; Piperazines ; therapeutic use ; Prognosis ; Proto-Oncogene Proteins c-kit ; genetics ; Pyrimidines ; therapeutic use

BACKGROUNDNominal atrioventricular (AV) interval in dual chamber pacemaker (DDD) is not the best AV delay in the majority of patients with atrioventricular block. To find a simple method for optimizing AV delay adjustment, we assessed surface electrocardiography (ECG) for optimizing AV delay during dual chamber pacing.

METHODSDDD pacemakers were implanted in 46 patients with complete, or almost complete, AV block. Optimal AV delay was achieved by programming an additional delay of 100 ms, to the width of intrinsic P wave or to the interval between pacing spike to the end of P wave on surface ECG. Left ventricular (LV) end diastolic and end systolic volumes, ejection fraction and diastolic parameters were measured by Doppler echocardiography during both nominal and optimal AV delay pacing.

RESULTSCompared to nominal AV delay setting, LV end diastolic volume increased [to (53.2 +/- 11.3) ml from (50.2 +/- 10.2) ml, P < 0.05], end systolic volume decreased [to (26.1 +/- 9.0) ml from (27.9 +/- 8.2) ml, P < 0.05] during adjusted AV delay pacing, resulting in an increase in LV ejection fraction [to (68.2 +/- 5.3)% from (64.5 +/- 4.3)%, P < 0.05]. LV diastolic filling and isovolumic relaxation time were not significantly changed.

CONCLUSIONOptimization of AV delay by surface ECG is a simple method to improve LV systolic function during dual chamber pacing.

Adult ; Aged ; Aged, 80 and over ; Atrioventricular Node ; physiopathology ; Cardiac Pacing, Artificial ; methods ; Electrocardiography ; methods ; Female ; Heart Block ; physiopathology ; therapy ; Humans ; Male ; Middle Aged ; Time Factors

This study was aimed to investigate the distribution of chromosomal aberrational karyotype in myelodysplastic syndrome (MDS) subgroups, the characterizations of numerical and structural aberration. The chromosome was prepared with simple culture of bone marrow, and the karyotype was analysed by G banding technique. The results showed tht 54 out of 127 patients (42.5%) had clonal chromosome aberrations, and the abnormal rates were different in subgroups: 30% (3/10) in MDS-RA, 35.9% (23/64) in MDS-RCMD, 22.2% (2/9) in MDS-RAS, 45% (9/20) in MDS-RAEB-I, 66.7% (14/21) in MDS-RAEB-II, 100% (3/3) in 5q-syndrome, respectively. Among 54 abnormal chromosome patients, 21 patients showed numerical aberration, 14 patients showed structural aberration, and the other 19 patients showed both numerical and structural aberration. The order of frequent aberrations was as follows complex karyotype (11.02%, 14/127), single +8 (10.24%, 13/127), -7/7q- (3.9%, 5/127), 1q+ (3.15%, 4/127), -X/-Y (3.15%, 4/127), 20q- (2.36%, 3/127), 5q- (2.36%, 3/127). The frequency of complex karyotype in MDS-RAEB (including RAEB-I and RAEB-II) was higher than that in non MDS-RAEB (including RA, RCMD, RAS, 5q-syndrome) (P < 0.05), and the frequency of balanced translocation was lower than that in non-balanced translocation (P < 0.05), and both of the two balanced translocation patients were found in MDS-RAEB. It is concluded that MDS is highly heterogeneous clonal disorder, a great majority of cytogenetic changes can be detected and most of which are recurrent aberrations, balanced translocations are rare, and only found in MDS-RAEB. The frequency of complex karyotype in MDS-RAEB is higher, and the patients with dup (1) (q21q32) recurrent abnormality is common in this study.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Chromosome Aberrations ; Cytogenetics ; Female ; Humans ; Karyotype ; Karyotyping ; Male ; Middle Aged ; Myelodysplastic Syndromes ; genetics ; Young Adult

OBJECTIVETo evaluate the value of normalization window of tumor vasculature (NWTV) in patients with unresectable gastric cancer undergoing neoadjuvant chemotherapy.

METHODSFrom October 2010 to March 2011, 93 patients with unresectable advanced or locally advanced gastric carcinoma were prospectively collected and randomly divided to Group A(n=30), Group B(n=29), and Group C(n=34). Group A received FOLFOX4 as conventional neoadjuvant chemotherapy. Group B received FOLFOX4 plus bevacizumab. The treatment was adjusted in Group C according to the hypothesis of NWTV with neoadjuvant chemotherapy delivered 5 days after bevacizumab treatment. The efficacy, drug toxicity and clinical outcome were assessed and compared between the three groups.

RESULTSThere were no significant differences among the 3 groups in demographics(P>0.05). All the patients completed the neoadjuvant chemotherapy. Efficacy and toxicity between the three groups were comparable(P>0.05). The rates of tumor downstaging in the three groups were 56.7%(17/30), 72.4%(21/29), 85.3%(29/34), respectively, with a significantly lower downstaging rate in Group C as compared to Group A(P<0.05). R0 resection rates were 23.3%(7/30), 27.6%(8/29), 52.9% (18/34), respectively, with significantly higher R0 resection rate in Group C as compared to Group A and Group B(All P<0.05). There was no perioperative death in this cohort. Postoperative complications were comparable among the 3 groups(P>0.05).

CONCLUSIONSAnti-angiogenesis agent can improve the efficacy of neoadjuvant chemotherapy in unresectable gastric cancer. Furthermore, administration according to NWTV may achieve better outcomes.

Adolescent ; Adult ; Aged ; Antibodies, Monoclonal, Humanized ; therapeutic use ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Bevacizumab ; Female ; Fluorouracil ; therapeutic use ; Humans ; Leucovorin ; therapeutic use ; Male ; Middle Aged ; Neoadjuvant Therapy ; Neovascularization, Pathologic ; drug therapy ; Organoplatinum Compounds ; therapeutic use ; Prospective Studies ; Stomach Neoplasms ; blood supply ; drug therapy ; Treatment Outcome ; Young Adult

BACKGROUND: Arthroscopic reconstruction for treatment of avulsion fracture at the tibial insertion of the knee posterior cruciate ligament (PCL) can minimize surgical trauma to the largest degree. However, its implants have relatively poor stability; therefore, functional exercise cannot be performed in the early stages after surgery, which is inconducive to knee function recovery. It is extremely challenging to perform traditional open reduction with internal fixation to repair avulsion fractures at the tibial insertion of the knee PCL. Often, the crushed bones cannot be firmly fixed, leading to a poor repair effect. OBJECTIVE: To design a new inverted "L"-shaped incision in the popliteal fossa through which bone plates were inserted to fix the crushed bones and to reconstruct PCL tension, facilitating knee function recovery; to compare the therapeutic effects of this new fixation and reconstruction method, and arthroscopic reconstruction for treating avulsion fracture at the tibial insertion of the knee PCL. METHODS: A prospective, single-center, non-randomized controlled trial. One hundred and eighty patients (knees) with avulsion fracture at the tibial insertion of the knee PCL will be assigned to two groups based on treatment methods: arthroscopic reconstruction group (n = 90;fracture fixation and repair under the arthroscope) and new method group (n = 90; bone plates will be inserted through an "L"-shaped incision in the popliteal fossa to fix the crushed bones and reconstruct PCL tension). After surgery, these patients will be followed up for 6 weeks, 6 months, and 12 months. RESULTS AND CONCLUSION: The primary outcome measure is the excellent and good rate of knee function recovery at 12 months after surgery as evaluated by Lysholm Knee Scoring Scale score (Herein referred to as Lysholm score). The secondary outcome measures are the excellent and good rate of knee function recovery before surgery, 6 weeks and 6 months after surgery; Lysholm score before surgery, 6 weeks, 6 months and 12 months after surgery; Hospital for Special Surgery (HSS) knee score, Visual Analogue Scale (VAS) score, posterior drawer test negative rate, X-ray morphology of the knee before surgery, and 6 weeks, 6 months, and 12 months after surgery and; incidence of adverse events at 6 weeks, 6 months and 12 months after surgery. Results of a preliminary study involving 62 patients (knees) with avulsion fracture at the tibial insertion of the knee PCL showed that posterior drawer test negative rate and Lysholm score were significantly higher in the new method group compared to the arthroscopic reconstruction group (P < 0.05) at 3 months after surgery. This study will be performed to compare the therapeutic effects of bone plate insertion through an "L"-shaped incision made in the popliteal fossa to fix the crushed bones and to reconstruct PCL tension, and traditional fracture fixation and repair under the arthroscope to treat avulsion fracture at the tibial insertion of the knee PCL. We believe that the former method will be superior to the latter one because it can fix the avulsion fracture more firmly, facilitating knee function recovery. This study was approved by Medical Ethics Committee of Cangzhou Central Hospital of China (approval No. 2017-120-01). This study will be performed in strict accordance with the Declaration of Helsinki formulated by the World Medical Association. Participants provided signed informed consent prior to participation in the study. This study was designed in December 2017. Patient recruitment and data collection will begin in April 2018. Patient recruitment will end in June 2019. Data analysis will be performed in August 2020. The study will be completed in October 2020. Results will be disseminated through presentations at scientific meetings and/or by publication in a peer-reviewed journal. This trial was registered with the Chinese Clinical Trial Registry (registration number:ChiCTR1800015026). The version of this study protocol is (1.0).

Osteoporosis is a metabolic bone disease characterized by decreased bone mass and degenerative changes in the microstructure of bone tissue, leading to increased bone brittleness and fracture risk. Bone fracture after osteoporosis is the most common and serious complication, which often leads to serious consequences in cases of inadequate prevention and late diagnosis. Therefore, more attention should be paid to prevention of osteoporosis and risk assessment of fracture and refracture after osteoporosis. This paper reviews the research progress in risk assessment of fracture and refracture after osteoporosis from the aspects of imaging, clinical manifestations and laboratory examination indexes. In recent years, the imaging methods have developed from dual-energy X-ray absorption, trabecular bone scoring and CT to high resolution peripheral quantitative CT; concern for their clinical manifestations has developed from independent risk factors to fracture risk assessment tools; the laboratory tests have developed from bone turnover markers and serotonin to microRNA. Although these developments have consistently increased the sensitivity of risk assessment for fracture and refracture after osteoporosis, problems still exist and need to be resolved.