Objective:To explore the feasibility of diagnosing glioma which expressing high level EGFR by 131I-EGF.Methods: In vitro cellular uptake and cellular retention experiment of ~(131)I-EGF was carried out with C6 cell.Radiodistribution experiment was carried out after injection of ~(131)I-EGF into 2-week-old tumor bearing Wistar rats,at 30min,2 hour,6 hour and 12 hour. SPECT images were carried out after 12 hours of intravenous injection of ~(131)I-EGF between tumor bearing rats and normal rats. Results:The rate of C6 cellular uptake and retention in vitro was high.At all selected time,~(131)I-EGF was mainly distributed in liver,spleen and kidney,partially distributed in tumor,but the radioactivity was stronger and stronger with the time passed,until reached the highest strong at 12h.Tumor tissues of tumor bearing rats were seen clearly by SPECT imaging after 12 hours of intravenous injection of ~(131)I-EGF,the liver and spleen were the major normal organs visualized on the images,and the other tis- sues didn't be seen obviously.Conclusions:~(131)I-EGF could offer a new and specific approach to diagnose tumor such as glioma and other tumors expressing high level EGFR from molecular level.

Objective:To review the recovery course of pituitary-thyroid axis by follow-up of hyperthyroidism patients with ~(131)Ⅰ therapy.Methods:This study involved 322 patients undergoing~(131)Ⅰ therapy between 1997 and 2000 in our hospital.Serum T_3,T_4 and TSH were regularly evaluated for 3 years after therapy.Results:Our study showed that serum T_3,T_4 and TSH returned to normal levels step by step,but the recovery of TSH fell behind that of T_3 and T_4.Conclusion:Our study demonstrates that clinical manifestations disappear in 1 year after ~(131)Ⅰ therapy,while the recovery of pituitary-thyroid axis takes more time.

The number of IPFI has increased in recent years because of the increasing of immunocompromised hosts due to abuse of antibiotics,glucocorticoids,immunosuppressants and all kinds of invasive operations,organ transplants.The mortality of IPFI remains highly because it's difficult to early diagnosis.Histopathologic biopsy remains as the gold standard for IPFI diagnosis although limited by materials.Imaging manifestations are nonspecific but have important suggestive values to some kinds of fungus.Direct examination is quickly but it's difficult to distinguish strain.Fungal culture can guide the treatment but its positive rate is low and the distinguish for contamination,infection and colonization is hard.Serological test has high sensitivity and specificity,but it cannot distinguish strain and it can yield false positives and false negative.Molecular biology is one of rapid developing methods,but its application is limited due to lack of standardized procedures and standards to assess its results,furthermore it can yield false positives.The proper diagnosis for IPFI relies on the comprehensive assessment combing with advantages and disadvantages of various methods.

BACKGROUND:Traditional total knee arthroplasty adopts medial parapatel ar approach, which induces severe trauma, requires long-term rehabilitation, and interferes the end point of quadriceps femoral tendon on superior patel ar pole. Total knee arthroplasty by subvastus approach has no impact on the knee-extension equipment, but it cannot provide sufficient exposure of surgical field and can induce damage to femoral muscle, so its application and safety need further exploration.
OBJECTIVE:To explore clinical effect of total knee arthroplasty by subvastus approach and medial parapatel ar approach.
METHODS:A total of 78 patients with 82 knees who were candidates for total knee arthroplasty were randomly divided into two groups. Treatment group (39 cases;41 knees) was given subvastus approach and control group (39 cases;41 knees) was given median parapatel ar approach. The knee function, range-of-motion of knee joint and complications after total knee arthroplasty were observed and compared.
RESULTS AND CONCLUSION:Compared with control group, the operation time in treatment group was significantly increased, while the postoperative wound drainage, straight leg raising time and walking time were obviously reduced (P<0.05). The scores of HSS before surgery and 12 weeks after surgery were not significantly different between the two groups (P>0.05), while at 1 and 6 weeks after surgery the scores in treatment group were significantly higher than those in control group (P<0.05). The range-of-motion of knee joint and MMT rank for quadriceps muscle were significantly improved in two groups after treatment (P<0.05), and those indicators in treatment group were more significant than the control group (P<0.05). The incidences of complications in treatment group and control group were 2.6%and 15.4%, respectively, with statistical y significant difference (P<0.05). The total knee arthroplasty by subvastus approach has less impact on the extensor mechanism, improves the recovery of knee function and range-of-motion of knee joint, and reduces the incidence of complications.

Objective To explore the distribution features of pathogenic spectra and antibiotic resistance of the isolates from blood cultures in hospital from June 2012 to June 2016.Methods A total of 4 238 blood samples from June 2012 to June 2016 were evaluated by BD Bactec FX-200,the identification results were used for retrospective analysis.Results A total of 455 positive pathogens were isolated from 4 238 blood cultures sample,the positive rate was 10.74％,Gram-positive accounts for 38.02％,Gram-negative bacilli accounts for 60.00％,Fungi accounts for 1.98％.Positive pathogens were distributed in newborn baby and middle-older patients,accounting for 6.78％ and 76.17％respectively.Which the Enterobacteriaceae accounting for 54.10％,the major consists were Escherichia coli and Klebsiellapneumoniae;Non-fermentative bacterial which consists of Pseudomonas aeruginosa and Acinetobacterbaumannii accounting for 2.90％.The major pathogens in Gram-positive cocci was Staphylococcus,accounting for 25.87％.Enterobacteriaceae were more sensitive to Meropenem,Imipenem and so all.Non-fermentative bacterial were more sensitive to Piperacillin/Tazobactam.Staphylococcus were more sensitive to Vancomycin and Linezolid.Streptococcus were sensitive to Vancomycin.Conclusion Combined with the distribution features of pathogenic spectra and antibiotic resistance,clinicians should pay attention to use of drugs reasonably to enhance the cure rate of bacteremia and Fungalemia.

Objective:To investigate the inhibitory effect of dihydroartemisinin on non-small cell lung cancer H1299 cells.Methods:Through the CCK-8 method for determining the IC 10 of dihydroartemisinin ,choose low dose IC 10 as the experimental concentration,CCK-8 to observe artemisinin in low doses of H1299 cell proliferation, cycle and the influence of radiation sensitivity.Results:IC10 of dihydroartemisinin was 14.87 μmol/L,dihydroartemisinin could inhibit proliferation of H 1299 cells,slow down the cell cycle , and increased the radiation sensitivity.Conclusion: Dihydroartemisinin can inhibit cell proliferation , cell cycle arrest in S phase ,increase the radiation sensitivity.

HepG2 cells were pre-incubated with insulin (Ins 0,1,0. 1,0.01 μol/L) and dexamethasone ( Dex 0,3,0. 3,0.03 μol/L) alone or together for 24 h to induce insulin resistance (IR) in vitro, the resistant level was estimated by glucose consumption, the optimal model of insulin resitance was chosen, and at the same time its lasting time of resistance was determined. In order to investigate the effects and mechanisms of mifepristone on in sulin-resistant HepG2 cells induced by insulin and dexamethasone, mifepristone and pioglitazone were adminis tered 24 h after the optimal model of insulin-resistant HepG2 cells was established. The glucose consumption, in tracellular concentrations of glucose, glycogen, ATP, and free fatty acid (FFA) in each group were detected. The expression of InsR-mRNA and GR-mRNA was detected by semi-quantitative reverse transcription and polymerase chain reaction (SqRT-PCR). Results revealed that pretreatment with insulin (0. 1 μmol/L) and dexamethasone (0.3 (μol/L) for 24 h caused optimal insulin resistance of HepG2 cells which lasted for 36 h. Compared with control group, the glucose consumption, intracellular glucose, glycogen, ATP contents and the level of InsR-mRNA in model cells decreased while FFAs concentrations and GR-mRNA increased. However, the tendency of insulin resistant HepG2 cells was obviously attenuated by pioglitazone at the concentration of 0. 2 mmol/L and mifepris tone at 200μmol/L and 20 μol/L while mifepristone at 2 μol/L had no effect on insulin-resistant cells. The findings indicated that mifepristone at 200 μol/L and 20 μol/L improved the insulin resistance via modulating intracellular glucolipid metabolism and the expression of InsR-mRNA and GR-mRNA.

10.3969/j.issn.2095-4344.2013.23.006

Objective Describes the use of homemade simple bedside bracket of children supracondylar fracture closed reduction and percutaneous pin fixed surgical methods and clinical effects.Methods 16 cases of children supracondylar fractures Gartland type Ⅱ cases,15 cases Gartland type Ⅲ,13 males and 3 females,with an average age of 6 years old,all patients used our hospital homemade simple bedside shelf assisted closed reduction,percutaneous pin fixation,and fixed with plaster immobilization.Results 16 patients were followed-up,in addition to the two cases of pin tract infection,fracture all were healed smoothly.Without iatrogenic nerve injury and internal fixation loosening,and also no volkmann contracture,and myositis ossificans etc complications.The average fracture healing time for three weeks,no bone delayed union or nonunion occurred.According to Flynn clinical function evaluation,there were excellent in 7 cases,good in 9 cases.Conclusion Useing homemade simple bedside shelf to children supracondylar fractures closed reduction and percutaneous Kirschner wire fixed can ensure the efficacy,and also simplify the surgical procedure,reduce radiation intake,which is the worthy of clinical practice.

Aim To construct human phage single-chain antibody library associated with esophageal cancer and to screen the specific scFv against Eca109 cells from the liberary. Methods Metastatic periesophageal lymph nodes of esophageal cancer patients were used as the B cells source, the total RNA of these B cells was extracted and prepared as the template of RT-PCR. First, we screened graticulely two pairs of primers of the heavy and light regions separately, then the V_H and V_L fragments were first amplified from the cDNA by the polymerase chain reaction (PCR). Second, the V_H-linker and V_L-linker were amplified from the V_H and V_L fragments. Last, the V_H-linker and V_L-linker were assembled into scFv gene fragments by SOE-PCR,and then Sfi I and Not I restriction site were inlet in it. ScFv gene was cloned into the pCANTAB-5E phagemid. Phagemids were introduced into E.coli TG1 by electrotransformation, followed by rescue of antibody-expressing phage using M13K07 helper-phage superinfection. Recombinant scFv phage library was constracted and PCR was used to identify the insert ratio of scFv antibodies library. Results of SfiI/Not I double digestion reaction positive insert clone were identified by 1.5% agarose gel electrophoresis. The phage library was panned with NHEEC and Eca109 cancer cells in suspension for four rounds. Strongly positive recombinant phage clones were used to infect E.coli HB2151. Expression of soluble scFv was induced by IPTG. Soluble scFv from periplasm were purified by affinity chromatography and identified by SDS-PAGE and Western blot. Cell ELISA , immunohistochemical staining and immunocytochemical staining were used to identify the activity of the soluble scFv. Results The result of agarose gel electrophoresis showed that total RNA of these B cells had two bands of 28 S and 18 S. The size of V_H fragment is about 450 bp,V_L fragment is about 350 bp and scFv is about 850 bp. The competence is 108 cfu??g-1 pUC18 DNA. Randomly digestive reac-tion showed that the positive insert ratio was 91.7% (22/24). After four rounds of panning, the fourth phage yield is 141 times as much as that of the first one. SDS-PAGE and Western blot showed that the MW of the soluble scFv was about 30 ku and the brand of 30 ku was stained. Immunohistochemical staining showed strong stainning of the tissue of esophageal cancer, but not the liver and gastric cancer tissue. Immunocytochemical staining showed significant staining of the esophageal cancer line Eca109. The result of cell ELISA assay revealed that soluble scFv had highly specific and could combined with Eca109 cells, but not with BGC-823 and NHEEC. Conclusion A human scFv phage display library associated with esophageal cancer has been constructed successfully and the specific scFv antibody against Eca109 has been identified from the liberary.