Objective To assess the prevalence of the A to G mutation at position 3243 of the mitochondrial tRNALeu(UUR) gene in type 2 diabetes in a Chinese population. Methods We screened 716 randomly selected, unrelated patients with type 2 diabetes for the mutation with a PCR-RFLP technique. Results Three individuals with this mutation were identified, representing approximately 0. 4％ of the type 2 patients screened. Further screening of first degree relatives of these 3 patients identified another 4 affected carriers. In comparison with type 2 diabetic patients without the mutation, these 7 carriers of the mt3243 mutation had:①an earlier onset of diabetes (38. 0±10.1 yr vs 53. 4±10.0 yr, P ＜0. 001) ;②lower BMI (19.5±2.0 vs 24. 9±10. 9, P ＜0. 0001) ;and ③ lower post-challenge insulin levels (Area under the curve of insulin levels during the OGTT, 2946± 1647.2 vs 7469±6647.7, P ＜ 0. 01). In addition, we screened the same 716 patients with type 2 diabetes, as well as 181 controls with normal glucose tolerance,for a newly described mt 3316 G→A mutation. This mutation was found in 16 patients with type 2 diabetes (2.20％) and 5 controls (2.7％). Therefore, the frequency of the mutation was not different between patients and controls.Moreover, clinical characteristics such as age of onset of diabetes, BMI, and insulin levels were not different between diabetic patients with the mt3316 mutation and those without it. Concision The mt3316 G→ A mutation is a polymorphism unrelated to diabetes.

AIM: To investigate the CE fingerprint of the compound Shengmai Powder(Radix et Rhizoma Ginseng rubra,Radix Ophiopogonis,Fructus Schisandrae chinensis). METHODS: Sequential uniform design was used to optimize the separation conditions. A CE fingerprint for Shengmai Powder was established using buffer comprising 44 mmol/L borate and 34 mmol/L SDS at pH 9. 5,a running voltage of 25 kV,a capillary temperature of 25 ?C and a wavelength of 200 nm. The sample was injected at a pressure of 50 mbar for 100 s. RESULTS: From the fingerprints of eleven batches of sample solutions,twenty main common peaks were determined. four peaks came from Radix et Rhizoma Ginseng,six peaks from Radix Ophiopogonis,thirteen peaks from Fructus Schisandrae chinensis,three peaks shared by Radix et Rhizoma Ginseng and Radix Ophiopogonis,one peak shared by Radix Ophiopogonis and Fructus Schisandrae chinensis and one new constituent. CONCLUSION: The developed method is accurate and reliable,and the fingerprint analysis can be used for the quality assessment and control of compound Shengmai Powder.

Objective To invstigate the effect of high dose X-ray irradiation on the expression of multidrug resistance-1 (MDR1), Bcl-2, MMP7 genes. Methods A nasopharyngeal carcinoma cell line,CNE1, were irradiated with a total dose of 50 Gy. The resistance to the cisplatin of CNE1 cells and the irradiated CNE1 (CNE1 R) cells was detected by MTT. mRNAs expression of MDR1 , Bcl-2 and MMP7 was measured by quantitative RT-PCR. Results The expression of MDR1 increased in CNE1 R cells. The semiquantitative A value of MDR1 mRNA was 0.47 ±0.04, and the value of CNE1R cells (1st, 7th, 21st,28th, 35th, 42nd and 49th days after irradiation) were 0.67 ± 0. 06 (t = -5.44, P = 0. 003) ,0.70 ± 0. 01(t=-5.90,P=0. 002),0.73±0. 01(t= -6. 45,P=0. 001) ,0. 67 ± 0. 03 (t= -3.97,P=0.011),0.65 ±0.01(t = -4.43,P=0. 007),0. 62±0. 05(t= -2. 64,P=0.046) and 0.62 ±0.02(t = -3.34,P=0.021), respectively. Bcl-2 mRNA expression were 0.55 ±0.02 and 1.05 ±0.04(t = -9.93,P=0. 000) and MMP7 mRNA expression were 0.51 ±0.01 and 0.82 ±0.02(t = -8.51,P=0.000) in CNE1and CNE1 R cells. Conclusions The MDR1 expression was increased after a total dose of50 Gy irradiation,which may be related to the synchronous change of Bcl-2 and MMP7 genes.

Objective To compare Western blot analysis and immunofluorescence method in Survivin expression. Methods By using Western blot analysis and immunofluorescence method, Survivin gene expressions of the peripheral blood mononuclear cells in 18 patients with leukemia and 10 healthy persons were analysed. Results Positive expression of Survivin gene was detected in 13 of 18 leukemic patients by Western blot analysis and in 11 of 18 leukemic patients by immunofluorescence method. Negative expression of Survivin gene was detected in 10 healthy persons. The results of Survivin expression were the same in 16 patients by Western blots analysis and immunofluorescence method (consistent rate is 88.89%), but the positive expression of Survivin by Western blot analysis was the negative expression by immunofluorescence method in another 2 patients. Conclusion Survivin gene expressed selectively in leukemic cells, but not in normal mononuclear cell in peripheral blood. It indicated that Survivin expression is effective monitoring index in diagnosis and prognosis of leukemia. Survivin expression by Western blot analysis was consistent with immunofluorescence method.

Objective:To clarify the relationship between epidermal growth factor receptor (EGFR) mutations and the clinicopathologic features of primary lung adenocarcinomas in Xinjiang.Methods:The mutations of EGFR gene at exons 18 -21 in 59 cases (including 15 cases of Uighur and 44 cases of Han) of lung adenocarcinoma tissues,which were obtained from surgical resection,were detected by amplifica-tion refractory mutation system (ARMS)method.And the relationships among mutations,race and clini-copathologic features were analyzed.Results:The frequencies of EGFR mutations in lung adenocarcinoma were 20% for Uighur,which was lower than that in Han (54.5%),P <0.05.The deletion mutations at exon 19 were seen in 2 of 15 Uighur cases and 9 of 44 Han cases.EGFR mutations were present,inclu-ding exon 21 L858R in one Uighur case and 12 Han cases,exon 18 G719X in two of 44 cases of Han, exon 21 L861Q in one of them.On histological type,the frequencies of EGFR mutation in alveolar pre-dominant adenocarcinoma was 71% (22 /31),which was higher than both that in solid predominant and mucinous carcinoma (6.7%,20% respectively).According to statistic analysis,EGFR mutations were without correlation with the patient’s gender,age,location,gross type,smoking status and lymph node metastasis(P >0.05).EGFR mutation was more frequent in well-differentiated cancer,mainly in acinar carcinoma,while poorly differentiated adenocarcinoma and mucous adenocarcinoma were lower.Conclu-sion:There was a difference of EGFR mutation in primary lung adenocarcinoma between Uighur and Han in Xinjiang,perhaps reflecting ethnic genetic variation,which is worth further analyzing.EGFR mutation was commonly detected in well or middle differentiated adenocarcinoma,mainly in acinar carcinoma.

Objective The increasing prevalence of infectious diseases is threatening the biological safety of donated blood in developing countries .This study was to analyze the prevalence of transfusion‐transmissible infectious related measures among first‐time ,voluntary blood donors from 1999 to 2013 in China .Methods From 1999 to 2013 ,all first‐time donors in the Xi′an Blood Service were screened for hepatitis B virus (HBV) ,hepatitis C virus (HCV) ,human immunodeficiency virus (HIV) and Trepone‐miapallidum (TP) and analyzed by trend test analysis .Results The positive rates of HBV ,HCV ,HIV ,and TP in the 415 657 blood donors were 1 .02% ,0 .55% ,0 .05% ,and 0 .46% ,respectively .The prevalence of HBV and HCV presented a decreased trend .Conclusion HBV infection is the primary threat to the blood safety ,while the increasing prevalence of TP and HIV might al‐so be a potential threat .

Objective: To observe the effect of acupuncture-moxibustion on the expression of insulin-like growth factor-1 (IGF-1) and suppressor of cytokine signaling-2 (SOCS2) in colonic mucosa of rat models of ulcerative colitis (UC), and explore the mechanism of acupuncture- moxibustion therapy in treating UC. Methods: The rats were randomized into a normal control (NC) group, a model control (MC) group, an herb-partitioned moxibustion (HPM) group and an electroacupuncture (EA) group, 8 in each group. The rat models of UC were established by immunological methods combined with local stimulation. The rats in the HPM and EA groups were given herb-partitioned moxibustion and electroacupuncture treatments respectively, once every day, lasting for 14 d. The morphological variations of rat's colonic mucosa were observed under light microscope; the colonic mucosal mucin was detected by PAS-AB and HID-AB staining methods; the expression of IGF-1 and SOCS2 was assayed by the immunohistochemical method. Results: In the rat models of UC, ulceration and inflammation of the colon were revealed by light microscope. The concentration of colonic mucosal mucin was reduced (P<0.01), while the expression of IGF-1 had an increase (P<0.01), and the expression of SOCS2 was reduced (P<0.01). After HPM or EA treatment, the pathological injuries of colonic mucosa had improved, the concentration of mucin increased (P<0.01), the expression of IGF-1 decreased (P<0.01), and the expression of SOCS2 increased (P<0.01). Conclusion: The secretion of mucosal mucin in rat UC decreased, the expression of IGF-1 was significantly higher, while the expression of SOCS2 was remarkably lower; both HPM and EA can help improve the damage of colonic mucosa in rat UC, and modulate the secretion of mucin, as well as regulate the expression of IGF-1 and SOCS2 in the colonic mucosa.

Objective To explore the application of PCR in detection and diagnosis of Echinococcus granulosus and develop a diagnostic technique by DNA hybridization labeled with DIG.Methods A pair of primers were designed according to Echinococcus granulosus gene fragment sequence, and the special primers were as: P1 5′-GGAATGGAGAGAAGTTAC-3′ P2 5′-GCAACCTCCGGAACTTGC-3′. The hydatid fluid, secondary hydatid and protoscolex of Echinococcus granulosus were used as template, 417 bp special band was got after being amplified by PCR. The PCR product was purified and labeled with DIG. The special DNA probe was successfully got and was used to detect the Echinococcus granulosus. Results The DNA from Escherichia coli, Shigella, Tubercle, Cysticercus granulosus and healthy human leukocyte were extended by PCR and hybrided by DNA probe labeled with DIG, and only Cysticercus granulosus expressed 471 bp special band. The sensitivity of PCR was that one Cysticercus granulosus can be detected or 100-10 fg DNA, while the dot hybridization was 2500 fg. Heterologous DNA did not be positive reaction, even increasing the spotting membrane dosages. Conclusion The technique of PCR labeled with DIG showed good specificity, high sensitivity, more accuracy and quickness and so it can provide scientific basis for early diagnosis and epidemiological investigation of Echinococcus granulosusEXPERIMENTAL RESEARCH WITH PCR AND DNA HYBRIDIZATION TECHNIQUE IN DETECTION AND DIAGNOSIS OF ECHINOCOCCUS GRANULOSUS$$$$ Feng Xiaomei, Ji Hugang, Cao Yinfang, Wang Wenhao (Department of Clinical Laboratory, Inner Mongolia Autonomous Region Hospital, Huhhot 010017, China) Abstract Objective To explore the application of PCR in detection and diagnosis of Echinococcus granulosus and develop a diagnostic technique by DNA hybridization labeled with DIG.Methods A pair of primers were designed according to Echinococcus granulosus gene fragment sequence, and the special primers were as: P1 5′-GGAATGGAGAGAAGTTAC-3′ P2 5′-GCAACCTCCGGAACTTGC-3′. The hydatid fluid, secondary hydatid and protoscolex of Echinococcus granulosus were used as template, 417 bp special band was got after being amplified by PCR. The PCR product was purified and labeled with DIG. The special DNA probe was successfully got and was used to detect the Echinococcus granulosus. Results The DNA from Escherichia coli, Shigella, Tubercle, Cysticercus granulosus and healthy human leukocyte were extended by PCR and hybrided by DNA probe labeled with DIG, and only Cysticercus granulosus expressed 471 bp special band. The sensitivity of PCR was that one Cysticercus granulosus can be detected or 100-10 fg DNA, while the dot hybridization was 2500 fg. Heterologous DNA did not be positive reaction, even increasing the spotting membrane dosages. Conclusion The technique of PCR labeled with DIG showed good specificity, high sensitivity, more accuracy and quickness and so it can provide scientific basis for early diagnosis and epidemiological investigation of Echinococcus granulosus[infections.

Child ; Child, Preschool ; Female ; Gastrointestinal Hemorrhage ; diagnosis ; Gastrointestinal Tract ; diagnostic imaging ; Humans ; Infant ; Male ; Radionuclide Imaging ; Sodium Pertechnetate Tc 99m

Objective To optimize the effective methods of isolation,purification,culture in vitro and identification of SD rat ovarian granulosa cells,to research the effects of Oviductus Ranae on the proliferation of rat ovarian granulosa cells by CCK-8,and to contrastively analyze the best optimal action concentration and time of serum contsining Oviductus Ranae on granulosa cells to lay the foundation for further in vitro experiment.Methods Nonage SD rats aged 25 d were selected and intraperitoneally injected by pregnant mare serum,then killed after 48 h.Ovarian granulosa cells were collected and cultured in the DMEM-F12 culture solution.The hematoxylin & eosin(HE) staining and immunofluorescence technique were used to identify the ovarian granulosa cells.Twen ty-five SD rats were randomly divided into the normal control group,positive medicine control group,and low,middle and high do ses Oviductus Ranae groups.Blood was collected and serum was separated after 7 d mediaction gavage.The volume percent of 10 ％,20％,40％,80％ serum in each group was added into the in vitro medium system of ovarian granulosa cells culture.Then the cell proliferation situation at 24,48,72 h in each group was measured by CCK-8.Results Oviductus Ranae significantly increased the proliferation ability of granulosa cells in a certain dose-dependent relation.With the increase of Oviductus Ranae concentration con centration,its.proliferation ability was gradually increased,after 48 h action,which in the Oviducthus Ranae-Comtaining serum group with the volume fraction of 20％ was most significant (P＜0.05).Conclusion Establishing in vitro cultural method of rat o varian granulosa cells is conductive to further research the action and mechanism of Oviductus Ranae on ovary.