According to the texts of Baihu Jia Renshen Decoction in the books of Maijing,Qianjin Yaofang,Qianjin Yifang,Taiping Shenghui Fang,Hxin Fang,and Jingui Yuhan Jing,the author drew the following conclusions:Baihu Jia Renshen Decoction in the present Shanghan Lun was very likely the written error Of Baihu Decoction,which was more close to the original meaning of Zhang Zhong-jing.From the period of Sui and Tang Dynasty,doctors had gotten new experiences in their practices.They added Ginsen into Baihu Decoction and named the new decoction"BaihU Jia Renshen Decoction",which was still in use today.Although there was not intrinsically difierence between Baihu Decoction and Baihu Jia Renshen Decoction,while the saying of"adding Renshen or not,should be decided by patient's Yuanqi"by doctor Shu Chi-yuan in Qing Dynasty had a significant impact.The erroneous views by some recent scholars can't be complied with,who advocated that the four main symptoms of Baihu Decoction syndrome should belong to syndrome of Baihu Jia Renshen Decoction according to the original texts in Shanghan Lun.

The article made a study on the earliest literature and the initial time of the "Five Orbiculi" theory in TCM. By analyzing the masons for different locations of "Five Orbiculi" recorded in ancient TCM books, the author argued against that the term of "Five Orbiculi" originated from ancient India and the "Five Orbiculi" theory was a product with the combination of traditioinal Chinese and Indian cultures. The author further put forward that the "Five Orbiculi" theory most probably was a Chinese traditional medical innovation under the influence of Internal Classic of Medicine, a great development to the theory recorded in Miraculous Pivot On Serious Confusion.

AIM: To observe the effect of E2F decoy DNA on proliferation and apoptosis of androgen-independent prostate cancer cell line PC-3M.METHODS: E2F decoy DNA,ARE decoy DNA and control decoy DNA were transfected into PC-3M cells with lipofectamine,respectively.Their effects on cell proliferation were detected by MTT assay.The changes of cell morphology were observed by inverted phase contrast microscope.The cell apoptotic rate was determined by flow cytometry(FCM) analysis and chromosome DNA ladder was detected by DNA gel electrophoresis.The expression of c-Myc mRNA and cyclin D1 mRNA was detected by RT-PCR.The protein levels of c-Myc and cyclin D1 were detected by Western blotting.RESULTS: The growth of PC-3M cells was inhibited after transfection.The transfected PC-3M cells displayed typical apoptotic morphological changes.The apoptotic rate was 26.35% and DNA ladder was observed after transfection.The expression of c-Myc and cyclin D1 were inhibited.CONCLUSION: These results indicate that E2F decoy DNA induces apoptosis of androgen-independent prostate cancer cell lines PC-3M and inhibits cell proliferation via inhibiting expression of c-Myc and cyclin D1.

AIM: To study the basic mechanism of transcriptional regulation, NKX3.1 gene promoter was cloned and its promoter activities in prostate cancer cell lines and other cancer cell lines were tested. METHODS: 1.04 kb - promoter fragment of NKX3. 1 gene was obtained by PCR and cloned into pGL3 - basic and pEGFP - 1 that are promoter - less reporter vectors to examine its promoter activity driving the reporter gene transcription. The promoter activity was determined by dual -luciferase reporter assay and the expression of GFP reporter observed under fluorescence micro scope. RESULTS: The sequence of the cloned 1.04 kb promoter proved to be correct by DNA sequencing. The dual - lu ciferase reporter assay (M1/M2) showed that the promoter activity in LNCaP cell transfected with pGL3 - 1.04 kb promoter was about 1.5 - fold higher than that of pGL3 - control transfection and 50 - fold higher than that of pGL3 - basic transfec tion. To investigate the 1.04 kb - promoter activity in different tumor cell lines, the constructed pGL3 - 1.04 kb promoter and pEGFP - 1.04 kb promoter were transfected into several cell lines, respectively. The results showed that the activity of 1.04 kb promoterin LNCaP was highest among the tested cell lines. Multiple consensus sequence elements have been iden tified within the 1.04 kb fragment using TRANSFAC database. Further experiments will be done to determine their founc tions. CONCLUSION: Cloned 1.04 kb fragment upstream of NKX3.1 gene presented a strong promoter activity and its activity was highest in LNCaP cell among the tested tumor cell lines.

AIM: To observe the effect of exogenous androgen responsive element decoy on the promoter of prostate specific antigen (PSA) and the growth of LNCaP cells for searching the possibility of gene therapy for prostate cancer. METHODS: Firstly, pGL3 - PSA luciferase expression vector containing 640bp - promoter fragment of PSA gene was constructed. Then, a 23 -mer phosphorothioated ARE decoy based on the deduced ARE sequence at the promoter region of PSA gene was synthesized. pGL3 - PSA and ARE decoy DNA were cotransfected into PC3 - M cell by lipofectamineTM 2000. Through detecting the activity of luciferase, the effect of ARE decoy on the promoter of PSA was studied. Then the ARE decoy DNA was transfected into LNCaP cells. The effect of decoy DNA on the proliferation of LNCaP cells was examined by using MTT assay. The effects of apoptosis were detected by phase contrast microscopy, DNA agrose gel electrophoresis and flow cytometry. Meanwhile, the nuclear extract was prepared from LNCaP cells and DNA - protein interactions were examined by electrophoretic mobility shift assay (EMSA). RESULTS: The reporter assay showed that the aetivity of luciferase was significantly reduced in the ARE decoy - transfected cells, bnt not in the cells transfected with the control decoy. EMSA demonstrated specific binding of the ARE decoy to androgen receptor. The growth of LNCaP was remarkably inhibited and apoptotic morphological changes as well as DNA fragmentation were observed in the ARE decoy- transfected cells. The rate of apoptosis was 22.4% detected by FCM. CONCLUSION: The ARE decoy is capable of inhibiting the promoter of PSA gene and inducing the apoptosis in prostate cancer cells. It may become a potential therapeutic tool for prostate cancers.

ObjectiveTo investigate the feasibility of examining aortic pulse wave velocity (PWV),aortic distensibility (AD) and brachial artery flow-mediated dilation (FMD) by means of highresolution 3.0 T MRI.MethodsA total of 32 healthy volunteers underwent high-resolution MRI to assess aortic PWV,and AD in ascending aorta (AA),proximal descending aorta (DA),distal descending aorta (DDA) and FMD of the brachial artery with repeat examination performed in 1-2 hours.PWV was evaluated by 2D Phase Contrast (PC) velocity-encoded MRI with a 4.7-7.8 ms temporal resolution.Fiesta-cine MRI was used to assess AD and FMD with a 18.75-31.25 ms temporal resolution.The image quality of these two scans was scored and the agreement between them was tested with Kappa analysis.The reproducibility of the results between repeated measurements of PWV,AA-AD,DA-AD,DDA-AD and FMD was assessed with intra-class correlation coefficient (ICC) analysis.The method of Bland-Altman plot was used to assess the agreement between results of repeated studies.Results Each examination including PWV,AD and FMD were completed in about half an hour.The image quality between repeated scans showed good agreement ( Kappa value 0.776 ) with the score of ( 3.53 ± 0.62 ) and ( 3.41 ± 0.67 ) respectively.Reproducibility between repeated measurements was high for aortic PWV [ (4.33 ± 0.88 ) vs ( 4.36 ±0.88) m/s],AA-AD [(8.60±3.11) × 10-3 vs (8.59 ± 3.10) × l0-3/mm Hg(1 mm Hg =0.133 kPa) ],DA-AD[ (6.95 ±2.44) × 10-3 vs (6.95 ±2.42) × 10-3/mm Hg],DDA [(10.54 ±2.91) ×l0-3 vs (10.55 ±2.90) × 10-3/mm Hg] and FMD [(24.94 ± 12.55)％ vs (24.92 ±1 2.38 ) ％ ].ICC were 0.95,0.97,0.99,0.98 and 0.94,P ＜ 0.01.Excellent agreement between repeated measurements was found for aortic PWV [ confidence interval (CI) between - 0.55 and 0.50 ],AA-AD ( CI between - 0.11 and 0.12 ),DA-AD ( CI between - 0.08 and 0.08 ),DDA-AD ( CI between - 0.23 and 0.21 ) and FMD (CI between - 1.46 and 1.51 ).The maximum difference percentage in minimum average for aortic PWV,AA-AD,DA-AD,DDA-AD and FMD was 38.53％,9.65％,3.86％,5.68％,42.37％,respectively,all less than 50％.Conclusion Comprehensive assessment of aortic compliance and brachial endothelial function can be achieved using 3.0 T high-resolution MRI with excellent reproducibility and within a reasonable amount of time.

Objective To investigate the imaging features of focal nodular hyperplasia and hepatocellular carcinoma on DWI. Methods The data of patients with histopathologically confirmed FNHs and HCCs between August 2008 and November 2010 were collected. A total of 24 patients with 26 FNH lesions and 36 patients with 39 HCC lesions were included in our study. All patients underwent breath-hold DWI with b = 500 s/mm2 and dynamic contrasted-enhanced (DCE) MRI. The imaging findings of FNHs and HCCs were retrospectively analyzed and compared. The signal intensity (SI) of the lesions on DWI were classified as iso-, slightly high, high SI and the distribution of SI between FNHs and HCCs was compared with Fisher exact test. ADC value and lesion-to-liver ADC ratio of FNHs and HCCs were measured and compared by using independent sample t test. ROC was performed to assess the diagnostic value of ADC value and lesion-liver ADC ratio in the characterization FNHs versus HCCs. Results Of 26 FNHs,23 manifested as isointensity or slightly high SI on DWI, but most 25 out of 39 HCCs showed high SI. The distribution of SI between FNHs and HCCs had significant difference ( P = 0. 000). The mean ADC value and lesion-liver ADC ratio for FNHs [ (1.76 ± 0. 62 ) × 10-3 mm2/s and 1.06 ± 0. 18, respectively ] were significantly higher ( P = 0. 001, P = 0. 000, respectively ) than those for HCCs [ ( 1.26 ± 0. 46 ) × 10-3mm2/s and 0. 79 ±0. 12, respectively]. The area (Az) under the ROC for the ADC value and lesionliver ADC ratio for the differentiation of FNHs versus HCCs were 0. 79 ± 0. 05 and 0. 85 ± 0. 05,respectively, with no significant difference (P =0. 270). The specificity of the two measures was 69. 23% and 97.44%, respectively, with significant difference (P = 0. 001 ). Conclusion FNH shows isointensity or slightly high SI with relatively higher ADC value and lesion-liver ADC ratio than those of HCCs on DWI,which is characteristic for its diagnosis and differentiation.

AIM: To study the basic mechanism of transcriptional regulation,NKX3.1 gene promoter was cloned and its promoter activities in prostate cancer cell lines and other cancer cell lines were tested.METHODS:(1.04 kb)-promoter fragment of NKX3.1 gene was obtained by PCR and cloned into pGL_3-basic and pEGFP-1 that are promoter-less reporter vectors to examine its promoter activity driving the reporter gene transcription.The promoter activity was determined by dual-luciferase reporter assay and the expression of GFP reporter observed under fluorescence microscope.RESULTS: The sequence of the cloned(1.04 kb) promoter proved to be correct by DNA sequencing.The dual-luciferase reporter assay(M1/M2) showed that the promoter activity in LNCaP cell transfected with pGL_3-(1.04 kb) promoter was about 1.5-fold higher than that of pGL_3-control transfection and 50-fold higher than that of pGL_3-basic transfection.To investigate the 1.04 kb-promoter activity in different tumor cell lines,the constructed pGL_3-(1.04 kb) promoter and pEGFP-1.04 kb promoter were transfected into several cell lines,respectively.The results showed that the activity of(1.04 kb) promoter in LNCaP was highest among the tested cell lines.Multiple consensus sequence elements have been identified within the(1.04 kb) fragment using TRANSFAC database.Further experiments will be done to determine their founctions.CONCLUSION: Cloned 1.04 kb fragment upstream of NKX3.1 gene presented a strong promoter activity and its activity was highest in LNCaP cell among the tested tumor cell lines.

Objective:To explore the clinical efficacy of radical resection for lung metastasis from colorectal cancer and the prognostic factors.Methods:The retrospective cohort study was conducted. The clinicopathological data of 63 colorectal cancer patients with lung metastasis who were admitted to Peking University Cancer Hospital from January 2004 to December 2015 were collected. There were 35 males and 28 females, aged (57±12)years. Patients underwent radical resection for primary lesion and lung metastasis from colorectal cancer. Observation indicators: (1) diagnosis and treatment; (2) follow-up and survival; (3) prognostic factors analysis. Follow-up was conducte by outpatient examination and telephone interview to detect the survival of patients after operation up to December 2018. Measurement data with normal distribution were represented as Mean±SD, and measurement data with skewed distribution were represented as M (range). Count data were described as absolute numbers or percentages. The Kaplan-Meier method was used to calculate survival rates and draw survival curves. Log-rank test was used for univariate analysis and COX proportional hazard model was used for multivariate analysis. Results:(1) Diagnosis and treatment: of 63 patients with lung metastasis from colorectal cancer, 6 had synchronous lung metastasis and 57 had metachronous lung metastasis. Eighteen cases of suspected lung metastasis were initially detected by chest X-ray, and further confirmed by computed tomography (CT). Forty-five cases of suspected lung metastasis were initially detected by chest CT. All the 63 patients underwent radical resection for primary and metastatic lesions. Two of 22 cases undergoing mediastinal lymph nodes dissection were detected one positive lymph node, respectively. All patients recovered well after operation, without severe complications. There were 57 of 63 patients receiving more than 6 months of postoperative adjuvant chemotherapy and targeted therapy based on fluorouracils. (2) Follow-up and survival: 63 patients were followed up for 8-143 months, with a median follow-up time of 58 months. During the follow-up, 19 of 63 patients died, 24 patients had secondary recurrence with a 5-year recurrence rate of 38.1%(24/63) and a recurrence interval of 18 months(range, 3-58 months). Of 24 patients with secondary recurrence, 19 had lung metastasis, 3 had brain metastasis, 2 had bone metastasis, 2 had liver metastasis; some patients had multiple metastases. Of 24 patients with secondary recurrence, 5 underwent reoperation and 19 underwent chemotherapy and radiochemotherapy. The 5-year overall survival rate of 63 patients was 62.7%. (3) Prognostic factors analysis: results of univariate analysis showed that location of primary lesion, the number of lung metastases and carcinoembryonic antigen (CEA) level before resection of lung metastasis were related factors for prognosis of patients with lung metastasis from colorectal cancer ( χ2=4.162, 7.175, 6.725, P<0.05). Results of multivariate analysis showed that the number of lung metastases and CEA level before resection of lung metastasis were independent influencing factors for prognosis of patients with lung metastasis from colorectal cancer ( hazard ratio=2.725, 2.778, 95% confidence interval as 1.051-7.064, 1.072-7.021, P<0.05). Conclusions:Radical resection for lung metastasis from colorectal cancer is safe and feasible. The number of lung metastases and CEA level before resection for lung metastasis are independent influencing factors for prognosis of patients with lung metastasis from colorectal cancer.

We constructed inducible and constitutive heterotrophic expression vectors of Dunaliella salina (D. salina) and identified heterotrophic transformants. A gene encoding a glucose transporter (Glut1) was cloned from human placenta tissues by RT-PCR and sequenced. Inducible heterotrophic expression vector pMDDGN-Bar of D. salina, which included a duplicated carbonic anhydrase (DCA) promotor and a Bar selectable marker that could drive expression of the Glut1 gene in D. salina, was constructed by molecular biology methods. In addition, we constructed another vector G5Glut1-Bar that contained a constitutive ubiquitin promotor, Glut1 and Bar box. The two expression vectors were introduced into D. salina by electroporation method, and then screened the transformants with phosphinothricin (PPT). Total RNA of the transformants extracted was used to analyze the integration of the target gene (Glut1) by RT-PCR. The cloned Glut1 sequence was 1479 bp and encoded 493 amino acids. The results of all enzymes digesting showed that two expression vectors were successfully constructed. After screening by PPT for several weeks, the transfomants grew well whereas wild-type cells died completely. The result of RT-PCR indicated that two transformants both had an about 250 bp specific band and the sequence homology was 100% compared with the human Glut1 sequence by Blast analysis. Taken altogether, inducible and constitutive heterotrophic expression vectors of D. salina was constructed successfully and the Glut1 gene was integrated into the genome of D. salina. Expression vectors above-mentioned may be used for the expression of the foreign Glut1 gene in D. salina.
Base Sequence ; Chlorophyta ; genetics ; metabolism ; Cloning, Molecular ; Electroporation ; Genetic Vectors ; genetics ; Glucose Transporter Type 1 ; biosynthesis ; genetics ; Industrial Microbiology ; Molecular Sequence Data ; Transformation, Genetic