Objective To perform the cytogenetic diagnosis on single lymphocyte of DMD patient with dystrophin gene exon 50 deletion.Methods Single lymphocytes of a DMD patient with dystrophin gene exon 50 deletion and normal volunteers were picked out and prepared for two-time duplex PCR.Results The rate of precise positive was 92%,91% and 93% in specimens of the patient(SRY positive,exon 50 negative),the male volunteer(SRY positive,exon 50 positive)and the female volunteer(SRY negative,exon 50 positive),respectively.Conclusion Two-time duplex PCR is fit for the genetic diagnosis of single lymphocyte from DMD patient with dystrophin gene exon 50 deletion.

OBJECTIVE To analyze the situation of extended spectrum ?-lactamases(ESBLs)and AmpC enzyme produced by nosocomial Escherichia coli isolates in 2005-2007.METHODS ESBLs were detected by double disk synergy test and disk diffusion confirmatory test.AmpC enzyme was detected by the three dimensional assay.Chi square test was used to test the significance.The application of different kinds of antimicrobials before the results of etiology be presented and the resistence rate of the ESBLs both producing and no producing were compared respectively.RESULTS The detectable rate of ESBLs in E.coli isolates of nosocomial and community infection was 55.1% and 21.3% and the detectable rate of AmpC enzyme nosocomial E.coli isolates was 17.4%.All strains were 100% susceptible to meropenem and imipenem but resistant to 15 other antimicrobials in different degree.The sensitivity to Piperacillin/tazobactam,cefoperazone/sulbactam and amikacin were relatively high.CONCLUSIONS The carrying rate of ESBLs from nosocomial E.coli isolates is high and AmpC enzyme and other resistance genes,which lead to multiple drug resistance.Standardized management of antimicrobials application should be strengthened and the consciousness of rational antimicrobials utilization should be raised.

Objective To study the moderate hypothermia with iced saline given intravenously with ice cap and ice sheet used in patients with severe stroke or craniocerebral trauma in order to clarify its clinical therapeutic effect and recovery of neurologic function for providing reliable clinical data.Methods A total of 84 patients suffered severe stroke (cerebral hemorrhage or cerebral infarction) or traumatic brain injury with GCS (Glascow Coma Scale) ＜8 admitted within 24 hours after onset were enrolled for study.The exclusion criteria were functional decompemsation of heart,liver,kidney and lung as cardiovascular complications of hypertension,hematological and other diseases,and craniocerebral trauma with multiple injuries of thorax,abdomen and extremities.Patients were divided into observation group and control group (n =42 in each group).The patients of two groups were managed to maintain respiratory tract patent,prevention of infection,dehydration,reduction of intracranial pressure and treated with Cytidine Diphosphate Choline,piracetam,ganglioside sodium and other conventional therapy.In control group,patients treated with ice cap and ice sheet,whereas in observation group,patients were treated with iced saline injected intravenously in addition to ice cap and sheet.At admission,the GCS score of patients was assessed and 6 months later,GCS score and Glascow Outcome Scale,(GOS) score of patients were determined,and 12 months after the treatment,quality of life score were evaluated by KPS.Results The time from onset of stroke or from trauma accident occurred to initiation of hypothermia treatment and time from initiation of hypothermia therapy to the target temperature reached in observation group were (8 ±4) hours and (3± 1 )hours respectively,and in control group,they were ( 16± 5) hours and (6 ± 2) hours,and there were significant differences between two groups ( P ＜ 0.05 ).The mean GCS score of patients at 6 months in the observation group and in the control group were (7.01 ± 1.12) and (4.02 ± 1.11 ) respectively (P ＜0.05 ) ; whereas the GOS score at 6 mouths of patients was categorized into good outcome,disabled outcome and poor outcome,and the good outcome in the observation group was significantly better than that in the control group (P ＜ 0.05 ),and no statistically significant difference in disabled outcome was found between two groups (P ＞ 0.05),and the poor outcome of GOS score was significantly lower in observation group than that in control group ( P ＜0.05 ).Twelve months after the treatment,the long-term quality of patients'life assessed by KPS score in observation group was significantly better than that in control group ( P ＜ 0.05).Conclusions The moderate hypothermia therapy by using iced saline administered intravenously combined with ice cap and ice sheet for severe stroke and head trauma patients conferred better therapeutic effects than the treatment with ice cap and ice sheet.

Objective To compare the effect of freeze-thaw tumor cells and the supernatant from tumor cell culture on the activation of lymphocytes. Methods Malignant melanoma B16 cells were prepared as tumor cell vaccine and the supernatant from tumor cell culture was collected at different time point.Transwell method was used to determine the chemotaxis of lymphocyte attracted by freeze-thaw tumor cells and the supernatant from tumnor cell culture. The cytotoxic activity of lymphocyte was detected by CCK-8 method. Results Freeze-thaw tumor cells and the supernatant from more than 2 h of tumor cell culture were found to show chemotaxis of lymphocyte. The chemotaxis of tumor cell culture more than 4 h was stronger than freeze-thaw tumor cells. Each group of chemotactic lymphocytes demonstrated to have the activity of killing tumor cells. The ability of killing tumor cells induced by the tumor cell culture more than 4 h was stronger than that induced by freeze-thaw tumor cells.In a certain range,the ability of lymphocyte chemotaxis and activation were enhanced over time. Conclusion The chemotaxis and cytotoxic activation of lymphocyte induced by the supernatant from tumor cell culture for a certain time are stronger than those by freeze-thaw tumor cells. The supernatant from tumor cell culture can be used as tumor antigen to get better immune activation instead of the freeze-thaw tumor cells.

Objective To investigate the impact of previous cystectomy for ovary benign cyst on ovarian reserve and pregnancy outcome in in vitro fertilization/intracytoplasmic sperm injection-embryo transfer (IVF/ICSI-ET) cycles. Methods Totally 622 infertility patients were retrospectively investigated who underwent first IVF/ICSI-ET cycle in Reproductive Hospital Affiliated to Shandong University from January 2013 to June 2014. There were 153 cases who had been removed ovarian cyst by cystectomy surgeries recruited as study group, in which 44 cases of ovarian endometriosis cyst, 35 cases of benign ovarian teratomas, 67 cases of simple ovarian cyst and 7 cases of ovarian mucinous cystadenoma. In contrast, 469 infertility patients with tubal-factor infertility or male factor were included as control group. The age-matched women in the control group had no ovarian surgery previously. The indicators of ovarian reserve and pregnancy outcome were analyzed between two groups. The influence of different types of ovarian cysts on ovarian reserve and pregnancy outcome in IVF/ICSI-ET cycles were also studied, ovarian endometriosis cyst was studied as Group A, and Group B consisted of benign ovarian teratomas, simple ovarian cyst and mucinous cystadenoma. Results (1) The significantly lower serum antimullerian hormone (AMH) level (median: 1.92 versus 2.90 mg/L), antral follicle count (AFC; median: 12.0 versus 13.0), retrieved oocytes (12 ± 5 versus 13 ± 6) and the number of embryo cryopreserved (median:1.0 versus 3.0) were found in study group compared with control group (all P<0.05). There was no statistical difference between two group for the following parameters, such as basal FSH level, the total dosage of gonadotropin duration and the total dosage of gonadotropin (all P>0.05). A better clinical pregnancy rate was achieved in control group (61.6%, 241/391) than that in study group (61.4%, 81/132), but no significant difference was existed (P=0.96). (2) Compared to Group B, Group A had fewer AFC, lower serum AMH level, retrieved oocytes and the number of embryo cryopreserved (11±4 versus 13±5;1.65 versus 2.15 mg/L;9±4 versus 13±5;0 versus 2.0;all P<0.01). There was a lower clinical pregnancy rate in Group A than that in Group B [50.0% (19/38) versus 66.0%(62/94)], accompanying with higher abortion rate [3/19 versus 9.7%(6/62)], but no differences were observed (all P>0.05). Conclusions Ovarian reserve declines after the cystectomy for ovarian benign cysts and the cystectomy has a negative impact on IVF/ICSI-ET cycle, resulting in a decrease of the number of retrieved oocytes and the number of embryo cryopreserved, but do not influence clinical pregnancy outcome. Ovarian reserve is impaired more seriously by cystectomy for ovarian endometriosis cyst than other ovarian benign cyst.

Objective To clarify the clinicopathological features of ovarian clear cell carcinoma derived from endometriotic cyst (EC-OCCC).Methods Totally 54 cases of EC-OCCC were recruited in the current retrospective study.The relation between ages, clinical symptoms and signs, surgical and pathological stages, serum CA125, findings of ultrasound, treatments and the sites of tumors, macro-and micro-features and expression of immunostainings were analyzed.Results (1) Clinical features: the ages of patients were (50±6) years old (range 31-62 years old).Pelvic mass was the major complaint of 50 patients (93％, 50/54).Forty-five cases belonged to International federation of Gynecology and Obstetrics (FIGO) stage Ⅰ, 4 cases were stage Ⅱ and another 5 cases were stage Ⅲ.Serum CA125 was elevated in 21 cases (54％, 21/39) before therapy.Doppler ultrasound showed 46 cases (85％, 46/54) had solid masses in pelvis.(2) Pathological findings: 52 cases (96％, 52/54) had their tumor unilaterally, and 2 cases (4％, 2/54) occurred bilaterally.The maximal diameters of endometriotic cyst (EC) ranged from 1.5 to 23.0 cm and maximal diameters of ovarian clear cell carcinoma (OCCC) components were from 0.5 to 12.0 cm.Fifty-one cases (94％, 51/54) had their tumor within EC, which showed focally irregular protrudings, grey-white papillae or solid nodules attached to the cyst wall.Three cases (6％, 3/54) appeared as irregular thickened wall of the cysts, ranged from 1.5 to 6.0 cm in the maximal length, with the microscopic features of EC and OCCC and the transitional areas between the 2 morphologies.All cases expressed cytokeratin (CK) 7 and pan-CK AE1/AE3, 17 cases (33％, 17/51) expressed ER and 5 cases (10％, 5/51) expressed PR.TP53 showed mutational phenotype in 19 cases (36％, 19/53).Sixteen cases (30％, 16/54) combined with uterine adenomyosis and 25 cases (46％, 25/54) with endometriosis at other sites.(3) Survival survey: during the period of 39.1 months follow-up, 3 cases relapsed and 2 cases died.(4) There was a significant difference of serum CA125 between patients of early-and advanced-stages (P=0.049).There were no differences identified in ages, diameters of EC and OCCC, the expression level of ER, PR and TP53, the co-existence of adenomyosis and endometrosis, as well as ultrasonic findings (P＞0.05).Conclusion EC-OCCC could be recognized in early stage by symptoms and ultrasound due to accompanied endometriotic cysts, resulting in relatively good prognosis.

Objective To describe the adequate feeding of early enteral nutrition in critical patients with cancer, and explore its influencing factors. Methods A retrospective analysis study was performed and 161 patients were involved. Results Nearly 57.14%patients got adequate feeding. The independent risk factors included complications (P < 0.05,OR=2.312,OR 95%CI 1.078-4.958), Acute Physiology and Chronic Health Evaluation Ⅱ (APACHEⅡ) scores (P < 0.05,OR=1.111,OR 95%CI 1.017-1.214) and hypoalbuminemia (P<0.05,OR=2.252,OR 95%CI 1.036-4.892) , and the protective factor was previous enteral nutrition experience (P<0.01,OR=0.123,OR 95%CI 0.021-0.494). Patients who got adequate feeding showed more increase in albumin levels, more decrease in APACHEⅡscores and shorter ICU stays. Conclusions Part of patients got inadequate feeding, due to comprehensive effects of many factors. And adequate feeding may be beneficial to the increase of albumin levels, also helps patients rehabilitate early and shorten the ICU stays.

Objective To confirm the clinical diagnosis of complete androgen insensitivity syndrome (CAIS) by molecular genetic testing in a large family. Methods PCR was performed to amplify the coding region of androgen gene, followed by direct sequencing in the patients with CAIS and relatives in this family. Results A missense mutation Arg773His was identified in the patients (homozygous) and carriers(heterozygous). Conclusions Mutation Arg773His in the AR gene leads to CAIS in this family. Molecular genetic testing of CAIS facilitates not only prenatal genetic diagnosis but also preimplantation genetic diagnosis and offers genetic counseling for future pregnancies to abandon the transmission of the mutated X chromosome to the coming generation.

Objective To determine the proper dosage of fentanyl for open heart surgery performed under cardiopulmonary bypass (CPB) without aortic cross-clamping and cardioplegia.Methods Twenty-seven ASA Ⅰ -H patients (10 male, 17 female) with fairly good cardiac function (NYHA Ⅰ - Ⅱ) scheduled for surgical repair of atrioseptal defect ( ASD) or ventricular septal defect (VSD) or mitral valve replacement (MVR) were studied. Age ranged from 18 to 44 years and body weight from 35 to 58 kg. The patients were randomly divided into three fentanyl dosage groups: group Ⅰ 10?kg ; group II 30 ?g kg-1 and group Ⅲ 50 ?g kg-1. Premedication consisted of intramuscular pethidine 1-2 mg kg-1 and scopolamine 0.05-0.06 mg kg-1 . Anesthesia was induced with midazolam 0.2 mg kg-1 and fentanyl 5 ?g kg-1 . Tracheal intubation was facilitated with vecuronium 0.15 mg g-1 . The patients were mechanically ventilated (Vr 8-10 ml kg-1 ,F 10-12 bpm,FiO2 100% ). The rest of the total dose of fentanyl (5 ?g kg-1 in group I , 25 ?g kg-1 in group II , 45 ?g kg-1 in group III) was infused after induction of anesthesia until the initiation of CPB, supplemented with inhalation of 0.6 % isoflurane. During CPB propofol was infused at 5 mg kg-1 h-1 . after discontinuation of CPB, again 0.6% isoflurane was inhaled until the end of surgery. Vecuronium 0.05 mg kg was given every 25-30 min during operation. EGG, HR, BP, CVP, SpO2, PET CO2 and body temperature (naso-pharyngeal and rectal) were continuously monitored during operation. Arterial blood samples were obtained before anesthesia (T0), 5 min after tracheal intubation (T, ) , immediately after thoracotomy (T2) , immediately before CPB (T3), 15 min after CPB was initiated (T4) , 10 min after termination of CPB (T5) and 5 min after chest was closed (T6) for blood gas analyses and determination of blood electrolytes and acid-base balance and blood concentrations of glucose, ACTH, angiotensin Ⅱ (A- Ⅱ ) and cortisol. Time of emergence from anesthesia and extubation were recorded.Results The demographic data, including age and body weight, CPB time and duration of surgery were comparable among the three groups. There were no significant changes in SpO2 , PETCO2 , body temperature, blood gases and electrolytes during operation in the three groups. MAP decreased significantly during CPB. The blood glucose, ACTH, A- Ⅱ and cortisol concentrations increased significantly during and after CPB as compared with the preanesthetic baseline (T0 ) ( P

Objective To investigate the feasibility of discriminating different solutions and solutions with different concentrations using gemstone spectral imaging (GSI).Methods Glucose solutions (GSs)with different concentrations (5%,10%,15%,20% g/mL)and 0.9% normal saline(NS)were scanned at a CT scanner with GSI model.Spectral analysis software was used to generate spectral curves of different solutions,representing as CT values on monochromatic images(40~140keV).The same procedure was repeated 3 days later.Reliability analysis with intra-class correlation coefficient (ICC)was used to evaluate the agreement between the twice CT scans.Nonlinear regression analysis was used to generate the regression equation of spectral curves of each solution.One-way ANO-VA was performed to compare the slopes,asymptotes and intercepts of each solution.Results Reliability analysis showed excellent agreements between the twice CT scans for each solution (ICC>0.9).The regression equation was expressed as:f(keV-40)=α-(α-β)×e [-(keV-40)×ρ],where f(keV-40)represented CT values on each monochromatic images,αrepresented asymptotes,βrepre-sented intercepts,ρrepresented slopes of each solution.Statistical differences were found among the slopes of NS and those of GSs with different concentrations (P <0.05),while no statistical differences were found among the slopes of different GSs (P >0.05). The result was highly suggestive that different ingredient solutions demonstrated different shapes of spectral curves.The asymptotes and intercepts of GSs with different concentrations were statistically different (P <0.05).The heights of the spectral curves of GSs elevated with the increase of the concentration.Conclusion Spectral curve with GSI model can be used to discriminate between NS and GSs,or even GSs with different concentrations.It provided a promising potential for in vivo body fluid analyses.