Objective To analyze the genetic polymorphism of three short tandem repeat (STR) double loci,namely,six STR loci in the Chinese population of Han nationality in Jilin district and to obtain the corresponding genetic data.Methods DNA was extracted from the blood cells of one hundred and three unrelated individuals of Han nationality in Jilin district;Six loci were divided into three groups which were groupⅠ(TPOX+CSF1PO),groupⅡ(D3S1358+D13S317) and groupⅢ(D5S818+D19S400).The extracted DNA were amplified with PCR multiplexing method.The PCR products were analyzed by non-reduced PAGE,followed by silver staining.The allele frequency distributions of six STR loci were studied by statistical procedures.The heterozygosity (H),discrimination power (DP),probability of exclusion (PE) and polymorphic information content (PIC) were calculated.Results The allele frequencies of six loci were obtained.The genotypes distributions of the loci were consistent with Hardy-Weinberg equilibrium and had higher heterozygosities and PIC.The H,DP,PE and PIC were 0.7511-0.8376,0.8273-0.9247,0.5167-0.6718 and 0.7100-0.8195 respectively.Conclusions The three STR double loci exhibited higher polymorphism and were better genetic markers.The data on the allele frequencies of these six STR loci may be used in individual identification,paternity testing and in other population genetic researches.

Objective To investigate the significance of the expression of endostatin and vascular endothelial growth factor(VEGF)in hepatocellular carcinoma(HCC).Methods The expression of endostatin and VEGF in 46 specimens which were collected at Second Affiliated Hospital of Fnjian Medical University from January 2000 to Apfil 2005 was detected by immunohistochemistry.The relationship between the expression of endostatin and VEGF and the progression of HCC was determined.All data were analyzed via t test,paired t test,Pearson rank correlation coefficient.LSD-t test or Tamhane's-t test.Results The expression of endostatin and VEGF was detected mainly in the cytoplasm of HCC cells and adjacent tissues.The values of mean optical density (MOD)of endostatin in HCC tissue,adjacent tissue and normal liver tissue were 0.11±0.02,0.14±0.0l and 0.09±0.01.respectively,and the values of integrated optical density(IOD)of endostatin in the above mentioned tissues were(1.8±0,2)×10~4,(3.8±2.2)×10~4 and(0.9±0.4)×10~4,respectively.The values of MOD and IOD of endostatin in HCC tissue were significantly lower than those in adjacent tissue(t=2.032,7.927,P＜0.05).The values of MOD of VEGF in HCC tissue,adjacent tissue and normal liver tissue were 0.13±0.02.0.12±0.02 and 0.11±0.02,respectively,and the values of IOD of VEGF were(5.4±3.1)×104,(3.9±2.5)×10~4 and(3.0 ±3.0)×10~4,respectively.The values of MOD and IOD of VEGF in HCC tissue were significantly hisher than those in adjacent tissue(t=5.871,8.723,P<0.05).There was positive correlation between the expression of endostatin and the recurrence of HCC(r=0.669,P<0.05),while VEGF had no influence on the recurrence of HCC(t=0.892,P>0.05).Conclusions Endostatin is mainly expressed in HCC tissue,but the level of its expression is lower in HCC tissue than in adjacent tissue.The expression of VEGF is higher in HCC tissue than in adjaneet tissue.Endostatin,rather than VEGF,may be used as a independent prognostic factor of the prognosis of patients with HCC.

ObjectiveTo explore the dynamic changes of electrocardiography (ECG) in patients with coronary heart disease(CHD) after the percutaneous coronary intervention(PCI). MethodsTo compare and analyze the regularity of the changes of 12-lead ECG on 38 cases in times,including one week,three weeks,one month,two and three months after the PCI. ResultsCompared with the ECG before the operation, the occurrences of the low-lying or the bidirectional T-wave were significantly increased,and lasting to 2 ～ 3 weeks after tie operation(5.0％ and 9.6％va2.6％ ,P ＜ 0.05). The ST segment depression( ＞ 1.0mm) and inverted T-wave in the 3 weeks and 2 months after the operation were significantly increased(5.0％ ,5.7％ vs 2.6％ ,P ＜ 0.05 ; 18.0％ and 25.2％ vs 16.2％ ,P ＜0.01 ). After that,the cases of ST segment depression were reduced to the level before PCI,and there was a significant reduction in the cases of the inverted T-wave ( P ＜ 0.01 ). ConclusionsIn the ECG of the patients with CHD after PCI, there were always early one-off abnormal Q-wave, changes of T-wave (inverted or low-lying, bidirectional)and ST segment depressions,etc. After that ,the changes above would turn back to the level before the operation.

Objective To evaluate the effect of intrathecal IL-10 gene on neuropathic pain induced by chronic constriction injury (CCI) to the sciatic nerve. Methods Sixty female SD rats weighing 230-250 g were anesthetized with pentobarfaital. Right sciatic nerve was exposed at the midthigh level and 4 ligatures were placed on the sciatic nerve with 4.0 catgut at 1mm interval. Intrathecal catheter (PE-10 tubing) were inserted at L5,6 interspace and correct placement was confirmed by outflow of CSF. The animals were randomized into 5 groups ( n = 12): group Ⅰ sham operation; group Ⅱ CCI; group Ⅲ,Ⅳ and Ⅴ received intrathecal (IT) normal saline (NS) (Ⅲ) or pcDNA 3.1 (Ⅳ) or pcDNA 3.1-IL-10 (Ⅴ) 3 days after CCI when the animals developed thermal hyperalgesia. Threshold to noxious thermal stimuli was measured before CCI (baseline), before and 2, 4, 7, 10 and 14 days after IT injection. CSF was collected on 1, 3, 7 and 10 days after IT injection for determination of CSF IL-10 concentration. Six animals were killed on 3rd and 14th days after IT injection respectively in each group and the sciatic nerve, lumbar segment of spinal cord ( L3-6) and hippocampus were isolated and blood was collected for determination of IL-10 mRNA expression (by RT-PCR) (on 3rd day after IT) and TNF-? content (on 14th day after IT) .Results Paw removal latencies were significantly longer 2-14 days after IT injection in group Ⅴ(CCI + pcDNA 3.1-IL-10) than in group Ⅲ (CCI + NS) and Ⅳ (CCI + pcDNA 3.1). The IL-10 mRNA expression in sciatic nerve, lumbar segment of spinal cord and hippocampus was significantly higher 3 days after IT injection while their TNF-? contents were significantly lower 14 days after IT injection in group Ⅴ than in the other 4 groups. The CNS IL-10 concentration on day 1-7 after IT injection was significantly higher in group Ⅴ than in the other 4 groups. Conclusion Intrathecal IL-10 gene injection can ease pain induced by CCI of the sciatic nerve through inhibition of inflammatory response.

Objective To explore the role of adjustable and close persistent peritoneal drain in large ascites and acute renal insufficiency(ARI)in patients with primary nephrotic syndrome(PNS).Methods The adjustable and close persistent peritoneal drain tube were planted in 6 cases with PNS accompanied by large ascites and ARI during December 2005 and August 2006.On the basis of drain,they were treated by prednisone,anticoagulative agent,hypolipidic agent,antibiotics,antioxidant,microcirculation-improving agent and aweto preparation.Results After the treatment,ascites disappeared or decreased.ARI was cured in 5 cases.Among these patients,partial remission(PR)and no respone(NR)to treatment were present in 3 cases respectively during hospitalization.After the follow-up of 3~12 months,the complete remisson(CR)of PNS was found in 2 cases,PR in 3 cases and NR in 1 case respectively.Conclusion The adjustable and close persistent peritoneal drain may be applicable for PNS patients with large ascites and ARI and slight glomerular lesion.It can clear away the superabundant water,mitigate the renal interstitial edema and accelerate the recovery of ARI.It also makes the patients have the opportunity of the treatment which includes predenisone,anticoagulative agent and hypolipidic agent.

Objective]To research the impact of Danxi’s medical science on Kampo medicine system and summarize its development in Japan,for providing ideas for the research of Chinese medicine. [Method]Learning Zhu Danxi related books and Kampo medicine literature, analyse the impact of Danxi ’s medical science on each genre of Japan Kampo medicine,explain the role of Danxi ’s medical science in the formation and development in Japan Kampo medicine system,and ummarize its development path in Japan.[Result]The development of Danxi ’s medical science in Japan experienced ups and downs, combined with local realities,and derived more Japanese-oriented medical theories. [Conclusion]Danxi ’s medical science not only for the formation of Japanese Kampo medicine system plays an important role, but also has a profound impact on Japan's three Kampo medical genre.Its development track is worth thinking and summarizing.

Objective To assess the efficacy of the mixtures of different concentrations of ropivacaine(R) and chloroprocaine(C)for epidural anesthesia in patients undergoing hysterectomy.Methods Sixty ASAⅠorⅡpatients aged 27-56 weighing 45-75 kg undergoing elective hysterectomy were randomly divided into 4 groups(n= 15 each);groupⅠ0.75% R alone;groupⅡ0.5% R-1% C;groupⅢ0.5% R-1.5% C and groupⅣ0.75% R+1% C.The epidural catheter was placed at L_(2,3)interspace and advanced 3.5 cm into the epidural space in cephalad direction.A total of 22 ml of epidural solution was injected in each group.The onset time,block height and duration of sensory block and the onset time,degree and duration of motor block(using Bromage scale)were assessed.The use of supplementary drugs(ketamine and ephedrine)and side effects were recorded.Results The onset time of sensory and motor block was significantly shorter in groupⅡ,ⅢandⅣthan in groupⅠ(0.75% R alone).The duration of sensory and motor block was significantly shorter in groupⅡandⅢthan in groupⅠandⅣ.The incidence of hypotension was significantly increased but the incidence of discomfort produced by traction on the viscera during operation was reduced in groupⅣas compared with groupⅠ.Conclusion The anesthetic efficacy of epidural 0.5% ropivacaine is significantly enhanced when mixed with 1.0% or 1.5% chloroprocaine.

Objective To investigate the effects of dihydrofolate reductase (DHFR) gene expression up-regulating on cisplatin resistance in epithelial ovarian cancer cell lines.Methods The cDNA length of DHFR gene was amplified by PCR and was connected to lentiviral vector pWPI, the recombinant retroviral vector DHFR-pWPI was infected SKOV3 cells by lipofectamine 2000.The groups included DHFR-pWPI-SKOV3 cell, pWPI-SKOV3 cell and SKOV3 cell group.Western blot was used to detect the expression of DHFR.Flow cytometry was applied to measure the cell apoptosis rate of 3 groups cells in different cisplatin concentrations (2.5, 5.0, 10.0, 20.0 μg/ml) and at different time period (24, 48 and 72 hours), and half maximal inhibitory concentration (IC50) treated with cisplatin concentration (6.0, 4.0, 4.9 μg/ml).High performance liquid chromatography (HPLC) was applied to test intracellular cisplatin concentration in different cisplatin concentration (4.0, 6.0, 8.0 μg/ml) at 24 and 48 hours.Transmission electron microscope was used to observe ultrastructural changes cells under IC50 cisplatin concentration.Results The recombinant plasmid DHFR-pWPI was constructed and then infected into SKOV3 cell successfully.(1) The expression of DHFR detected by western blot in transfection group was higher than those in the negative control group and blank control group (10.280±0.009 vs 2.050±0.003 vs 3.480±0.003;P＜0.01).(2) Treated with cisplatin concentration (2.5, 5.0, 10.0, 20.0 μg/ml) at 24, 48 hours, the apoptosis rate detected by flow cytometry results shown that they were lower than those in the negative control group and blank control group (P＜0.05), while treated at the concentration of 5.0 and 10.0 μg/ml for 72 hours, whose apoptosis rate in transfection group was higher than those in the negative control group and blank control group (P＜0.05).When treated cells under IC50 cisplatin concentration (6.0, 4.0, 4.9 μg/ml) at for 24 and 48 hours, the results indicated thatthere were mainly G0/G1 stage cell cycle rate in 3 groups, it was obviously higher in transfection group than those in two control groups (P＜0.05).However, mainly G2/M, S stage cell cycle rate for 72 hours, and S stage cell cycle rate in transfection group obviously higher than those in two control groups, but G2/M stage cell cycle rate were lower (P＜0.01).(3) After treated with cisplatin concentration (4.0 μg/ml) for 24, 48 hours and cisplatin concentration (6.0 μg/ml) for 24 hours, the intracellular cisplatin content tested by HPLC method in the transfection group were significantly lower than those in two control groups (P＜0.01).While, at 6.0 μg/ml of cisplatin concentration for 48 hours and 8.0 μg/ml of cisplatin concentration for 24 and 48 hours, the intracellular cisplatin content of transfection group were obviously higher than those two control groups (P＜0.05).(4) Treated with IC50 (6.0, 4.0, 4.9 μg/ml) cisplatin concentration at different time to obeserve ultrastructural changes by transmission electron microscopy.The results shown that the microwire gathered together at 24 and 48 hours, and the number and structure of mitochondria had obvious change in transfection group, while there was rare microfilament, the number of mitochondria decreased but structure change was not apparent in two control groups.There were appeared expansion of endoplasmic reticulum and had rare normal organelles among three groups.After treated with cisplatin for 72 hours, there were inordinate microfilament, a part of nuclear membrane disappeared, a lot of ribosomes gathered together in two control groups, and there were rare microfilament in transfection group, nuclear membrane completely disappeared, many white cystic matter were seen in cytoplasm, mitochondrial structure disappeared completely, which seems most cells on the verge of death.Conclusion The lentiviral expressing vector harboring human DHFR gene were successfully constructed.When the up-expression of DHFR gene, the drug-resistant in ovarian cancer cell may be increase, which suggest that there were certain contact between resistance increases with microfilament gathered and the change of the mitochondria.

Background and purpose:Dihydrofolate reductase (DHFR) is expressed highly in platinum-resis-tant ovarian cancer. This study aimed to explore the relationship between the silence ofDHFR gene and platinum drug resistance in ovarian cancer, and lay the foundation for the treatment of platinum-resistant ovarian cancer.Methods:To design targeting hairpin siRNA ofDHFR gene, the optimal siRNA silent sequence was selected, and lentiviral vector carryingDHFR gene was constructed successfully, named DHFR-pGCSIL-SKOV3 cell. Flow cytometry was used to detect the cell apoptosis of DHFR-pGCSIL-SKOV3 cells, pGCSIL-SKOV3 cells and SKOV3 cells incubated in various concentrations of cisplatin (2.5, 5.0, 10.0 and 20.0 μg/mL) at different time points (24, 48 and 72 h), and cell cycle changes of these cells at IC50 cisplatin concentration (4.4 μg/mL). High performance liquid chromatography was used to test intracellular concentration of cisplatin at different induction concentration of cisplatin (2.5, 5.0 and 7.5 μg/mL) and various time points (24 and 48 h). Ultrastructural changes of these cells at concentration of cisplatin IC50 (4.4 μg/mL) were observed by transmission electron microscope.Results:After annealing double-strand nucleotide was connected to pGCSIL/GFP vector, sequencing result was correct. SKOV3 cell were transfected with virus particles followed by Western blot detection of interference effect. Flow cytometry was used to detect apoptosis in three groups of cells, and increased apoptosis rate was found at the raised cisplatin concentration (2.5, 5.0, 10.0 and 20.0 μg/mL) at 24, 48 and 72 h in DHFR-pGCSIL-SKOV3, pGCSIL-SKOV3 and SKOV3 cells. The apoptosis rate in DHFR-pGCSIL-SKOV3 was signiifcantly higher than that in pGCSIL-SKOV3 and SKOV3 cells at 24 and 48 h (P<0.05). Flow cytometry was adopted to test cells cycle of 3 groups at different time period under IC50 cisplatin concentration (4.4 μg/mL), the results indicated that G0/G1 phase cell rate of DHFR-pGCSIL-SKOV3 was much more than the others, of which G2/M and S phase cell rates were on the contrary. While at 72 h, 3 groups were mainly G2/M and S phase cell rates, DHFR-pGC-SIL-SKOV3 was lower than the others. High performance liquid chromatography method was used to detect intracellu-lar cisplatin concentration at 24 and 48 h after the cells were incubated at various concentrations of cisplatin (2.5 and 5.0μg/mL). The results showed the intracellular cisplatin content of DHFR-pGCSIL-SKOV3 cell was signiifcantly higher than that of pGCSIL-SKOV3 and SKOV3 cells. However, after incubation at cisplatin concentration of 7.5 μg/mL, the intracellular cisplatin content of DHFR-pGCSIL-SKOV3 cell was signiifcantly lower than that of pGCSIL-SKOV3 and SKOV3 cells at 24 h, while higher than pGCSIL-SKOV3 and SKOV3 cells at 48 h (P=0.034,P=0.014). We observed ultrastructural changes of three different cell lines induced by IC50 cisplatin concentration(4.4 μg/mL) at different time points by the electron microscope. We found that the microiflaments were increased and gathered together and mitochondrial structure was also changed obviously without the drug. However, there was rare microiflament in three groups of cells at 24 and 48 h, while at 72 h, obviously increased inordinate microiflaments were observed.Conclusion:We successfully constructed pGCSIL lentivirus interference carrier carryingDHFR gene. The research indicates that down-regulation ofDHFR gene is related to cisplatin drug resistance in ovarian cancer. The results laid the foundation for us to investigate the molecular mechanisms of multidrug-resistance in tumor.

Objective To analyze and compare the independent and combined diagnostic values of high frequency ultrasonography (HFU) and molybdenum target imaging (MTI) for early breast tumor in a prospective study.Methods 258 patients with breast mass which were suspected as breast tumors by physical examination received HFU,MTI,their combined applications and operative treatments.Comparative studies were made between the surgical pathology and imaging evaluations.Results Totally 258 cases were enrolled in this study.Surgical pathology confirmed 135 cases of early breast cancer and 123 cases of benign breast tumor.For the diagnosis of early breast cancer,combined application was more effective than HFU or MTI alone in sensitivity (P<0.05),and more superior than MTI alone in accuracy.While the specificity,positive predictive value,negative predictive value, false positive rate, false negative rate had no difference among HFU, MTI and combined applications (P>0.05).In the patients of early breast cancer, compared with HFU,MTI found less nodules of calcification(18.3% vs 36.5%,P<0.01),but revealed more axillary lymph metastasis (69.0% vs 33.3%,P<0.01).Conclusion HFU and MTI alone or their combined application are reliable imaging methods for the diagnosis of early breast cancer.For the sensitivity and accuracy of diagnosis, combined applications has more superior advantages.