Objective To observe the clinical curative effect of sijunzi decoction composite with shenqi pills in the treatment of spleen and kidney yang deficiency syndrome in patients with primary hypothyroidism(hypothyroid-ism).Methods 62 spleen kidney yang deficiency patients with hypothyroidism were randomly divided into 30 cases in the control group and 32 cases in the treatment group.Patients in the control group were given L-thyroxine tablets orally,once a day;patients in the treatment group were given sijunzi decoction combined with shenqi pills for decoc-tion,daily 1 agent,3 times a day orally,on the basis of the control treatment group.Two groups treatment cycle were 4 months,and the clinical efficacy of the two groups were observed.Results The treatment group total effective rate was 96.9%,which was higher than that in the control group 80.0%(χ2 =4.402,P<0.05);before and after treat-ment,TCMsyndrome scores were significantly decreased in the two groups (t=21.40,25.40,all P<0.01)and treatment group decreased more significantly(t=4.30,P<0.01 ).Before and after treatment,TSH,FT3,FT4 in the two groups were significantly improved (control group:t=5.31,1.14,4.13,treatment group:t=6.27,1.85,6.08,all P<0.01),and the degree of improvement in treatment group was more obvious compared with the control group (t=1.33,0.67,278,all P<0.05).Conclusion Sijunzi decoction and shenqi pills can significantly improve the clinical symptoms of primary hypothyroidism of spleen-kidney yang deficiency type in addition to thyroid function and blood lipid,indicating that this method is capable of achieving satisfactory clinical effects in treating primary hypothyroidism of spleen-kidney yang deficiency type.

Objective To explore the effects of the therapeutic effects of interventional heated chemotherapy( IHC) for moderate and advanced huge primary hepatocellular carcinoma and compare with transcatheter arterial chemoembolization( TACE) . Methods The 68 pa-tients with moderate and advanced huge primary hepatocellular carcinoma accepted the treatment of IHC were considered as the observation group,and 32 patients managed TACE were regarded as the control group. The therapeutic effects,living quality,secondary surgery rates,The reduced level of serum AFP,survival rate,and toxic side effects of 2 groups were contrasted. Results The effective rate of observation group was obviously higher than that of control group (79. 4% vs 56. 3%,P <0. 05),and secondary surgery rates was increased(70. 6% vs 43. 8%,P<0. 01). The ratio of increased Karnofsky score was more than 20 in observation group,which was obviously higher than that of control group(35. 3% vs 12. 5%,P<0. 01). T The reduced level of serum AFP was more than 50% in observation group,which was higher than that of control group (52. 9% vs 25. 0%,P<0. 01). The survival rate of 12,18,24 months after treatment in control group was in-creased than that in control group individually (P<0. 01). Conclusion IHC is effective for huge primary hepatocellular carcinoma and in-creases the secondary surgery rates,which is one of the safe and effective methods for huge primary hepatocellular carcinoma.

Objective To inhibit the expression level of SALI4 in AML cell line THP-1 and investigate its potential effects on pathogenesis of leukemia. Methods AML cell line THP-1 was transfected with plasmids that expressed small interfering RNA targeting SALL4. The samples were divided into 4 groups:(1) blank group: samples with not any treatments; (2) control group: cells with empty pRS vector alone;(3) test1 group:cells with SALL4-shRNA-pRS-1 plasmid transfection complex; (4) test2 group:cells with SALL4-shRNA-pRS-2 plasmid transfection complex. The expression levels of SALL4 mRNA and protein were measured by real time fluorescence quantitative PCR and WB. C-myc, Cyclin D1 and β-catenin were important components of Wnt/β-catenin signaling pathway and their expression levels in SALL4 knockdown THP-1 cells were detected by real-time fluorescence PCR. Furthermore, THP-1 apoptosis was analyzed by flow cytometry after Annexin V-PI staining. Results Real time fluorescent quantitative PCR illustrated that the expression of SALL4 in testl group, test2 group, control group and blank group were ( 36. 0 ± 4. 3 ) %,(32. 0 ± 2. 4) %, ( 102. 0 ± 6.5 ) % and ( 100. 0 ± 2. 6 ) % respectively. There was statistical significance ( F = 226. 3, P < 0. 05 ). The expression of SALL4 in testl and test2 group respectively were significant lower than that in blank group (t = 19.7,19. 1, P<0. 05). The expression of SALL4 had no significant difference between blank group and control group (t = 1.1, P ＞0. 05). Western blot analysis revealed SALL4 protein in testl and test2 group were significantly decreased compared with those of control and blank group. All above data indicated the high efficiency of RNA interference targeting SALL4. Comparing with the blank group, the relative expression of C-myc, Cyclin D1 and β-catenin mRNA in test1, test2 and control group were(44.0 ±6.2)%,(44.0 ±5.1)% and (107.0±13.6)%;(22.0±4.5)%,(25.0±3.5)% and (48.0 ± 7. 6 ) %; ( 42.0 ± 3.5 ) %, ( 59. 0 ± 3.7 ) % and ( 79. 0 ± 5.6 ) %. The expression of C-myc,β-catenin and Cyclin D1 mRNA in testl and test2 group were significant lower than that in blank group (t = 10. 1,9. 5, 23. 3, 22. 9; 17.4, 12. 4; P < 0. 05). The percentage of apoptotic cells in group of test1,test2,control, blank were (57.2 ±9.1)%, (34.4 ±8.6)%, (14.4 ±3.6)% and (14.8 ±4.8)%respectively. There was statistical significance ( F = 42. 5, P < 0. 05 ). After the inhibition of SALL4, the percentages of apoptotic cell in testl and test2 group were significantly increased( t =9. 7, 4. 5 ;P <0. 05).Conclusion The inhibition of SALL4 in leukemia cell line THP-1 downregulates the expression of cell proliferation related genes such as C-myc, Cyclin D1,β-catenin and promoted apoptosis.

Objective To investigate the effect of health education on motor function in hemiplegia after stroke. Methods 105 stroke patients were divided into treatment group （53 cases）and control group（52 cases）. The patients in both groups received the similar rehabilitation and medicines, and the patients in the treatment group also accepted the health education. They were assessed with the Fugl-Meyer assessment （FMA） and modified Barthel index （MBI）. Results The scores of FMA and MBI in the both groups were improved after treatment（P<0.001）, and the result of the treatment group was better than that of the control group （P<0.05）. Conclusion Health education can improve the recovery of motor function and activity of daily living in patients with hemiplegia by knowing more about the disease.

Objective To explore the capability of Ang-1 gene delivery to acute myocardial infarction using targeted microbubbles carrying ICAM-1 antibody.Methods Thirty-seven rabbits' left circumfles branch coronary arteries were ligated for models.Three rabbits were injected with microbubbles carrying ICAM-1 to detect the ability of targeting.Thirty-four rabbit models were divided into 3 groups randomly as follow:IM group (n=12,accept direct intramuscular injection),ICAM-1 group (n=12,accept intravenous injection of targeted microbubbles and Ang-1) and control group (n=10,without any treatment).Ultrasonography were executed on all animals before and 2 weeks after the treatment.All rabbits were killed after 2 weeks and examined for Ang-1 mRNA and protein by RT-PCR and Western-Blot respectively.Microvessel density (MVD) counting of infracted myocardium,observed by factor Ⅷ immunochemical staining,was performed to value the proangiogenesis effect of Ang-1 delivered by targeted microbubbles carrying ICAM-1 antibody.The liver and the kidney in ICAM-1 group were taken to assess the systemic delivery.Results IM and ICAM-1 group showed significantly improvement in the ejection fraction (P＜0.05) while control group did not.Ang-1 mRNA and protein could be detected in IM and ICAM-1 group;however,the expression between the two groups showed no siginificant difference.None of the control animals showed Ang-1 expression.compared with ICAM-1 group,the MVD was greater in IM group.Ang-1 was not detected either in liver or in kidney in ICAM-1 group.Conclusions Targeted microbubbles carrying ICAM-1 antibody can deliver Ang-1 gene to ischemic myocardium directly.Meanwhile,it's as effective as IM injections besides the greater angiogenesis effect.This strategy improves the perfusion of acute myocardial infarction and the function of heart.

Objective To certificate the effects on transfection ratio and cells viability of ultrasound (US) acoustic intensity, radiation duration, microbubbles concentration and DNA concentration in delivery human angiopioetin-1 gene (hAng-1) into 293T cells by SonoVue microbubbles and decide the optimal transfection parameters. Methods Mix 293T cells and SonoVue microbubbles linked with eGFP-C_3-hAng-1 in a different way, detect the gene transfection ratio and cells viability under the various US intensity, radiation duration, microbubbles and DNA concentrations. Results The gene expression would be increased if enhanced the intenstiy of US,radiation time,microbubbles and DNA concentrations,and the cells viability would be kept more than 90% ( P <0. 01). Whereas,if the US intensity increased over 1. 5 W/cm~2 ,the duration over 30 s and microbubbles and DNA concentrations over 20% and 15 mg/L respectively,the gene expression would not increase significantly ( P > 0. 05),whereas coupled with obviously decreased cells viability( P <0. 01). Conclusions The optimal conditions of deliver hAng-1 gene into 293T cells by SonoVue microbubbles was mixing cells and microbubbles in a cell wall-sticky way,US intensity was 1. 5 W/cm~2, duration 30 s,20% microbubbles and 15 mg/L DNA concentration.

Objective To investigate the transfection efficacy and expression of Ang-1 gene and proangiogenesis in vitro and vivo by ultrasound-mediated microbubble destruction.Methods 293T cells were divided into three groups:group A was given hAng-1 plasmid and microbubbles plus ultrasonic irradiation,group B was given hAng-1 plasmid and ultrasound,group C was given hAng-1 plasmid only (without ultrasound).Forty-eight hours after transfection,the transient expression rate was observed under fluorescence microscopy and flow cytometry.RT-PCR and Western blot analysis were taken to evaluate the mRNA and protein expression of Ang-1 respectively.Twenty-seven rabbit models of ligated left circumflex branch coronary artery were divided into 3 groups randomly as follow:group Ⅰ (accepted intravenous injection of SonoVue microbubble and Ang-1 plus ultrasonic irradiation),group Ⅱ (accepted intravenous injection of Ang-1 with ultrasound),group Ⅲ (control group).Myocardial contrast echocardiography (MCE) was executed on all animals before and after the treatment.Two weeks after gene delivery,RT-PCR and Western blot analysis were taken to evaluate mRNA and protein expression of Ang-1 respectively.Microvessel density (MVD) counting of infracted myocardium,observed by Factor Ⅷ immunochemical staining,was performed to value the proangiogenesis effect of Ang-1 delivered by ultrasound mediated cavitation of microbubble.Results Green fluorescence was observed in group A and B by fluorescence microscopy,which was negative in group C.The transfection expression rate was significantly improved in group A ( P ＜ 0.01).In vivo,Microbubbles could be observed in former ischemic myocardium in MCEexamination and the Ang-1 mRNA and protein could be detected in group Ⅰ.On the other hand,the contrast agent was defected obviously and none of the animals showed Ang-1 mRNA and protein expression in other two groups.The MVD counting showed significant improvement in group Ⅰ whereas other two groups didn't.ConclusionsMicrobubble-enhanced ultrasound exposure can improve the Ang-1 gene transfection expression rate observably both in vitro and in vivo.This strategy for delivering has great proangiogenesis effect in vivo.

Objective:To evaluate the value of ultrasound-guided transperineal systematic prostate biopsy(SPB)and cognitive fusion multi-parameter magnetic resonance imaging(mpMRI) suspicious transperineal targeted biopsy(CFTB) in the prostate cancer with different serum prostate specific antigen(PSA) levels.Methods:A retrospective analyses were performed in 527 patients with suspected prostate cancer who underwent ultrasound-guided SPB from January 2018 to December 2019 in Shanghai Jiaotong University Affiliated 6th People′s Hospital. According to the PSA levels, they were divided into group A(PSA 4-10 μg/L) and group B(PSA>10 μg/L). All the patients underwent ultrasound-guided SPB, 376 patients with suspicious mpMRI had two additional targeted biopsies. The detection rates of ultrasound-guided SPB and CFTB in prostate cancer were tested by χ 2 test. Compared with pathological results, the sensitivity, specificity, accuracy of two methods were calculated and tested by χ 2 test, and a P<0.05 was defined as statistically significant difference. Results:Prostate cancer was detected in 319 of 527 patients(60.5%). One hundred and three cases of 198 patients in group A were diagnosed as prostate cancer, with an overall detection rate was 52.0%. Among them, ultrasound-guided SPB detected 72 cases of prostate cancer, the detection rate was 36.4%, sensitivity was 67.9%, specificity was 17.7%, accuracy was 26.5%, the detection rate, sensitivity, specificity and accuracy of CFTB were 39.9%, 75.6%, 91.6% and 88.8%, respectively. In this group, there were no statistically significant differences in the detection rate and sensitivity of the two methods in the diagnosis of prostate cancer (χ 2=0.525, 0.005, both P>0.05), and the differences in specificity and accuracy were statistically significant (χ 2=108.340, 79.829, respectively, both P<0.05). Two hundred and sixteen cases of 329 patients in group B were diagnosed as prostate cancer, with an overall detection rate was 65.7%. Among them, 160 cases of perineal prostate cancer were detected by ultrasound-guided SPB, with the detection rate was 48.6%, sensitivity was 78.2%, specificity was 37.6% and accuracy was 49.5%. A total of 189 cases of prostate cancer detected by CFTB, the detection rate was 57.4%, the sensitivity was 89.3%, the specificity was 90.6%, and the accuracy was 90.2%. All the differences were statistically significant in group B(χ 2=5.131, 4.391, 61.339, 38.982, all P<0.05). Conclusions:When PSA is greater than 10 μg/L, CFTB has a higher diagnostic efficiency than SPB.When PSA is 4-10 μg/L, there are no significant differences between the two methods in the detection rate and sensitivity of prostate cancer.

Objective To investigate the appropriate setting up of normal reference ranges of lymphocyte subsets in some flow cytometry laboratories and to study the effects of different flow cytometers and various reagents by different manufacturers on the analysis of peripheral blood lymphocyte subsets. Methods Three FCM labs (named A, B and C) in Beijing region were selected representing 3 commonly used flow cytometers (Beckman Coulter Epics XL, Beckman Coulter Cytomics FC500, BD FACS Calibur). 50 samples from healthy donors were distributed to 3 labs and tested according to individual lab's standard operating procedure to verify whether the normal reference ranges of peripheral blood lymphocyte subsets established were appropriate. The application of internal quality control was also investigated. Commercial blood quality control reagents were given to the 3 FCM labs and tested within 20 working days paralleled with routine samples. In addition, 20 patients' samples were prepared using 4 different combinations of reagents ( a , b , c and d). The results from combination a, which used the Beckman Coulter reagents and instrument, were compared to the results from combination b, c and d, which used reagents from different manufacturers. Then the prepared samples were tested on Beckman Coulter Epics XL to evaluate the effects of different combinations of reagents on the results of peripheral blood lymphocyte subsets analyzed by the same instrument. Furthermore, 24 patients' samples prepared by same reagents from Beckman Coulter company were tested on both Beckman Coulter Epics XL and BD FACS Calibur respectively to assess the effects of different instruments on peripheral blood lymphocyte subsets. 20 patients' samples prepared by same reagents and instruments were analyzed by Beckman Coulter Epics XL analytic system and BD FACS Calibur analytic system respectively to assess the effects of the two analytic systems on the lymphocyte subsets. Results Over 10% of the results for NK and T4/T8 in lab A as well as T4 in labs B and C fell outside of their normal reference ranges. The probabilities exceeding corresponding normal reference ranges were 16% ( 9/50 ), 24% ( 12/50 ), 22% (11/50) and 12% ( 6/50 ), respectively. The results using internal blood quality control in 3 FCM labs within 20 working days were all within the reference ranges of the quality control provided by the kit. The biases from b and c reagent combinations were substantial compared with that of reagent a combination. Among the biases from b and c reagent combinations, the lowest probability of bias exceeding 10% was T8 of combination b, which had probability of 70% (14/20). The highest probabilities of hias exceeding 10% were T3 and T4 of b and c reagent combinations, which reached 100% (20/20) . Furthermore, the biases of T3, T8 and B of d reagent combination compared with that of reagent a combination were also substantial. The probabilities of bias exceeding 10% were 35% (7/20) ,85% (17/20) and 75% (15/20), respectively. Comparing the results of samples prepared and analyzed by reagents and instruments from different manufacturers to that of samples prepared and analyzed by the same company's reagents and instruments showed that there were great discrepancies in T3, T4 , T8 , B and NK. The probabilities of bias exceeding 10% were 71% ( 17/24), 80% (19/24) ,38% (9/24), 33% (8/24) and 92% (22/24), respectively. The biases of T8, NK and B were substantial when compared the results from Beckman Coulter Epics XL analytic systems and BD FACS Calibur analytic systems. The probabilities of bias exceeding 10% were 55% (11/20 ), 70% ( 14/20 ) and 55% (11/20), respectively. Conclusions FCM labs should set up their own normal reference range for peripheral blood lymphocyte subsets. The normal reference range should be verified periodically. It is important to apply internal blood quality control regularly and accumulate the quality control results. The reagents and instrument for preparing peripheral blood samples should be from the same manufacturers.

Objective To test the efficiency of gene transfer and expression mediated by ultrasound and microbubble strategy in 293T cell. Methods SonoVue microbubbles were mixed with pla.smid DNA encoding hAng-1 and green fluorescent protein. The mixture of the DNA and microbubbles was administer to cultured 293T cells by ultrasound exposue. The ultrasound condition was 1. 5 W/cm~2 and 30 s. Forty eight hours later, transfer rate was assessed by fluorescence microscopy and flow cytometry. Cell viability was assayed by Trypan Blue staining. RT-PCR and Western blot analysis was used to examine the expression of hAng-1 mRNA and protein. Agarose gel electrophoresis was used to evaluate the integrity of the plasmid. Results The transfection expression rate of eGFP in 293T cells was markedly increased with the additon of 20% microbubbles and 15 mg/L DNA. Fetal calf serum had no influence on the gene transfer rate. Ultrasound irradiation combined with microbubbles couldn't destroy the integrity of plasmid. Conclusions Ultrasound-mediated microbubbles destruction can increase the transfection and expression of hAng-1 gene in 293T.