Diterpenes, an important class of natural compounds, are widely distributed in nature. As the valuable diterpenoids continue to be found, diterpene synthase in the course of diterpene synthesis get as much attention as possible. The multiformity of end-product-diterpenoids were also due to the diversity of diterpene synthase. This paper focuses on the advances in recent biosynthesis pathway of diterpene and types, cloning, catalytic mechanism, synthetic biology application.

Objective Partial hepatectomy, liver transplantation, and radio frequency ablation for hepatocellular carcinoma (HCC) were compared to select the most suitable method for HCC. Methods 1198 patients with HCC in 3 hospitals in Guangzhou were divided into 3 groups: group Ⅰ , small HCC; group Ⅱ > HCC without vascular invasions and group Ⅲ , HCC with vascular invasion. The patients either received partial hepatectomy, transplantation or ablation. The 1-, 2- or 3-year survival rates, the 3-year recurrent rates and Child-Pugh grades in the 3 groups were compared. Results For small HCC, there was a significant increase in the 3-year survival rate (P<0. 05) and a significant decease in the recurrent rate (P<0. 05) in patients who received transplantation, compared with those who received hepatic resection. Patients who received ablation had a higher 3-year survival rate and a lower recurrence (P<0. 05) in comparison with those who received hepatectomy. There was no significant difference(P<0. 05) between transplantation and ablation, but there were more Child A patients who received hepatectomy and ablation, and more Child B and C patients who received transplantation. For advanced HCC, there was no significant different in the 3-year survival rates for the 3 therapies, but the 3-year recurrence was lower (P<0. 05) in the transplantation group. Conclusions For small HCC, superiority of transplantation versus resection was obvious. Ablation (diameter <3 cm) was also superior to resection, whereas ablation was as effective as transplantation. There were more Child B and C patients in the transplantation group than the ablation and resection groups. Therefore, small HCC with hepatic decompensation should receive liver transplantation. Transplantation was advantageous in having less tumor recurrent but there was no difference in the 3 therapies for advanced HCC.

Objective Radiosensitivity is highly correlated with DNA doub le stra nd breaks(DSBs). The repair of human DSB is possible mainly through nonhomologou s DNA end joining(NHEJ)pathway. This study was designed to determine the relat ionship between expression of 70Ku/ 80Ku prot ein (a modulating subunit of DNA-PK, which is an important component in NHEJ pathway) and radiosensitivity of nasoph aryngeal carcinoma cell lines. Methods Radiosensitivity parameters of the nasoph aryngeal carcinoma cell lines were obtained by the colony forming assay and radi ation dose-survival curve was done by linear quadratic model. Western blot was p erformed to determine the expression of 70Ku and 80 Ku in total extract of CNE1 a nd CNE2 at baseline level、different time after 4?Gy irradiation、12 h after di ff erent doses of irradiation. To perform a semi-quantitated assay of protein expr e ssion, the optic density (OD) value of each band was estimated by automatic imag e analysis system. Results Surviving fraction of CNE1 was higher than those of CNE2 at each dose point. Mean inactivation dose was higher in the CNE1(2.35)t han that in the CNE2(1.11), but the quantity of 70Ku/ 80Ku in either cell line was similar at both baseline and postirradiation levels. Conclusions CNE2 is mo re radiaosensitive than CNE1; as there was no marked correlation observed betwee n the quantity of Ku protein and radiosensitivity of the nasopharyngeal carcinom a cell lines. X-irradiation cannot result in any change in total quantity of Ku protein.

Objective To set up the molecular cytobiological model of endogenous coagulation factor Ⅷ (FⅧ) re-expressing in human liver cells L02,and to study the regulation pathway and molecular basis of the re-expression of FⅧ in L02 cells activated by NO signal.Methods The L02 cells at logarithm growth phase were selected and randomly divided into blank control group and experimental group, inhibitor group and inhibitor control group;they were cultured for 0,12,24,36,48,and 60 h.Flow cytometry was used to detect the expression of human FⅧ protein in L02 cells after treated for 48 h.Griess experiment was performed to detect the levels of NO in L02 cells at different time points;the transcription levels of human FⅧ gene,iNOS gene,NF-κB1 gene and I-κB alpha gene were detected by RT-PCR method.Western blotting method was used to detect the expression levels of human phosphorylated I-kappaB (phosphorylated I-κB)in L02 cells.Results The results of flow cytometry showed that the expression of human L02 FⅧ protein was found after treated with L-arginine for 48 h. The Griess results showed that the levels of NO in L02 cells in experimental group were significantly increased at 3,6,12,and 24 h (P<0.05)and the levels of NO in blank control group,inhibitor group and inhibitor control group had no changes. The RT-PCR results showed that the transcription of human FⅧ mRNA in L02 cells was found in experimental group,but there was no transcription of human FⅧ mRNA in blank control group,inhibitor group and inhibitor control group;the transcription levels of iNOS,NF-κB1 and I-κB alphain experiment group were increased(P<0.05)and the transcription levels of these genes in blank control group,inhibitor group and inhibitor control group had no changes. The Western blotting results showed that after adding L-arginine the expression level of phosphorylated I-κB was significantly increased (P < 0.05 ), other groups had no such change. Conclusion L-arginine can activate the phosphorylation of I-κB by NO signal pathway to lead to the changes in the expression of human FⅧ gene promoter upstream regulatory-related transcription factors NF-κB to activate the expression of human FⅧ in human liver cells L02.

Objective To study the pathogenesis of upper gastrointestinal rehaemorrhagia after the transjugular intrahepatic portasystemic shunt (TIPS) and its influencing factor.Methods Fifty postoperative patients with TIPS were selected.The patients were followed-up,and the effect of the various factors in the role of upper gastrointestinal rehaemorrhagia after TIPS was analyzed.Results The portal vein pressure of 50 patients with TIPS decreased from preoperative (39.8 ±9.2) cmH2O (1 cmH2O =0.098 kPa) to postoperative (25.2 ± 5.8) cmH2O,and there was statistical difference (P ＜ 0.05).Fourteen patients appeared upper gastrointestinal rehaemorrhagia after TIPS,which accounted for total of 28％ (14/50) and included 3 cases of postoperative vomiting blood within 3 days.Acute stomach mucosa lesions bleeding was considered,and bleeding was controlled within a short-term medical treatment (1 patient after more than a year in recurrent upper gastrointestinal rehaemorrhagia).Twelve cases of patients appeared upper gastrointestinal rehaemorrhagia within 2 years after TIPS,and the causes of rehaemorrhagia in 6 cases were esophageal variceal rehaemorrhagia,gastric and duodenal ulcer in 3 cases,erosive gastritis in 2 cases,coagulation abnormalities in 1 case.Esophageal variceal rehaemorrhagia rate was 12％ (6/50).Conclusions The main reasons of upper gastrointestinal rehaemorrhagia after TIPS are variceal rehaemorrhagia and non variceal rehaemorrhagia,both of which are important causes of rehaemorrhagia after TIPS.Variceal rehaemorrhagia after TIPS occurs more than 3 months,and non variceal rehaemorrhagia occurs within 3months,so it is very important to protect gastric mucosa with proton pump inhibitor in postoperative patients.

Objective To compare the sensitivity of HE,immunohistochemistry (IHC) and RT-PCR in detection of breast cancer metastases in axillary lymph nodes.Methods Twenty female patients with newly diagnosed and clinically node-negative breast cancers underwent modified radical mastectomy, including a complete axillary lymph node dissection. The ages of the patients ranged from 31 years to 65 years, and the diagnosis of breast cancer was approved by pathological finding. Two hundred and thirty-nine axillary lymph nodes were found in these 20 patients. Metastases in axillary lymph nodes were explored by HE, cytokeratin 19 IHC and RT-PCR for cytokeratin 19 respectively. Results Seven(2.9%) lymph nodes were found to have metastatic cancers by HE in 3 patients,all nodes were found in level Ⅰ. Metastatic cancers were found in 13 (5.4%) nodes by IHC in 7 patients,11 nodes in level Ⅰ and 2 nodes in level Ⅱ; and 52(21.8%) nodes were found to contain tumor cells by RT-PCR in 14 patients,30 nodes in level Ⅰ and 22 nodes in level Ⅱ. All of 7 histologically(HE) positive nodes were found to contain tumor cells by IHC and RT-PCR. Among 232 histologically(HE) negative nodes,6(2.6%) nodes were found to contain tumor cells by IHC,and 45(19.4%) nodes were found to contain tumor cells by RT-PCR, all 6 IHC positive nodes showed the expected 460-base pair products on gel electrophoresis ( P

Objective To evaluate the effects of the short-term intervention lifestyle intervention on metabolic measurements of community patients with impaired glucose regulation (IGR). Methods A total of 90 IGR participants were randomly assigned to the control group (n=45) or the intervention group (n= 45). The subjects in the control group received routine diet and physical exercise advice once a month. The subjects in the intervention group received additional individualized diet counseling and circuit-type resistance training. Metabolic parameters were compared before or after the intervention between the two groups. Results In oral glucose tolerance test (OGTT),2-h plasma glucose (PG) and homeostasis model of insulin resistance index (HOMA-IR) were significantly decreased in the intervention group at 3 months(F= 13.47 or 82.25 ,both P < 0.05). Body mass index (t=-2.44, P<0.05), systolic blood pressure (t= -3.39, P<0.05), diastolic blood pressure (t=-3.97, P<0.05), fasting plasma glucose (t=-3.89, P<0.05),2-h PG (t=-7.22,P <0.05) ,total cholesterol (t=-2.72,P<0.05),low-density lipoprotein cholesterol (t=-2.74, P<0.05), and glycosylated hemoglobin A1 C (t=-3.73, P<0.05) were significantly declined in the intervention group compared to the control group (all P<0.05). Conclusions Intensive lifestyle intervention can significantly improve the metabolic markers of IGR subjects and should be used to prevent type 2 diabetes.

Objective: To observe the effects of acupuncture on synovial pathology, synovial mast cell degranulation and tryptase expression and to investigate the relationship between the functions of mast cells and effects of acupuncture on early adjuvant arthritis in rats. Methods: Forty-six male Wistar rats were randomly divided into normal control group (n=16), untreated group (n=15) and acupuncture group (n=15). Adjuvant arthritis was induced by injection of 0.1 mL Freund's complete adjuvant in right hind limb footpad. Normal control group and untreated group received no acupuncture treatment, while rats in the acupuncture group were treated with sterilized disposable stainless steel needles inserted perpendicularly as deep as 2 to 3 mm at Xuanzhong (GB39), 6 mm at Shenshu (BL23) and 7 mm at Zusanli (ST36) for eight times (15 min each time) every two days. Setting the modeling day as the 0 day of the experiment, the body weight and paw volume of the rats were measured every three days from the 0 day. In the end, synovial tissues of the right hind ankles were sampled and made into paraffin sections. Then they were firstly stained with hematoxylin-eosin for observing synovial pathology to evaluate the effects of acupuncture on adjuvant arthritis, then stained with toluidine blue for observing the number and degranulation ratio of synovial mast cells and finally detected by immunohistochemical staining method to investigate the expression of tryptase in synovium. Results: Compared with the untreated group, the body weight of rats in the acpuncture group was increased significantly (P<0.05), while the paw volume decreased obviously (P<0.01). Hematoxylin-eosin staining showed that acupuncture significantly inhibited inflammatory cell infiltration, synovial cell hyperplasia, and synovial fibroplasia in synovium of rats with adjuvant arthritis as compared with the untreated group (P<0.05). Toluidine blue staining showed that acupuncture could significantly diminish the numbers of total and degranulated mast cells in rats with adjuvant arthritis (P<0.01), which were significantly higher in the untreated group than in the normal control group (P<0.01). Showing by immunohistochemical staining, the expression of tryptase in synovium in the acupuncture group was decreased as compared with the untreated group (P<0.01). Analyzed by Spearman's bivariate correlation, the number of mast cells and degranulation ratio of mast cells were positively correlated with the pathological scores (r=0.837, P<0.01; r=0.634, P<0.01). Conclusion: Acupuncture can improve pathological condition of inflammatory synovium in rats with early adjuvant arthritis by inhibiting the function of synovial mast cells, which may play an important underlying role in the immunoregulation of acupuncture on adjuvant arthritis.

Objective To investigate the MRI imaging features, and pathologic basis of the orthotropic transplantation nude mouse model with human pancreatic cancer. Methods Adopting Siemens Magnetom Trio Tim 3.0 Tesla superconductive MRI and breast coil was used to examine 30 orthotropic transplantation nude mouse models of the human pancreatic cancer, these mouse were sampled to acquire TSE-T1 -weighted and T2-weighted transverse axial images. Intraperitoneal injection of Gd DTP A was used to perform continuous dynamic enhancement scanning. Signal intensities of tumors were measured in plain scanning and each phase' s enhancement scanning images, respectively. Intensification rates of tumors were calculated. Pathologic examination of tumors was performed to be compared with the findings of MRI scanning. Results The successful rate of inoculation of 30 nude mice was 100%. The histological findings were comparable with poorly differentiated adenocarcinoma. Compared with signal of adjacent tissues, the MRI findings of the tumors were uniformly slightly hypointensity (90% , 27/30) , or unevenly (10% , 3/30) on TSE-T1WI; uniformly (20% , 6/30) or unevenly (80% , 24/30) hyperintensity with equal or more hyper signal spots on TSE-T2WI. Signal intensities on plain scanning was 228.35 ±11.71, and 1.5,3,6,9, 12 min after enhancement scanning, thesignal intensities were 258.20 ± 11.17, 301.75 ± 17.09, 358.65 ±25.13, 480.05 ± 19.01, 558.35 ± 40.49, which were significantly higher than those in plain scanning (P <0.01). The intensification rate of every phase was 0.13 ±0.04, 0.35 ±0.11, 0.56 ±0.10, 1.10 ±0.10, 1.45 ±0.18, and the difference among these phases was statistically significant (P <0.01). The significantly intensified area was the area where the tumor cells grew actively with rich capillaries; the central area without intensification was the area of necrotic tissue and/or densely packed tumor cells and few capillaries. Conclusions High resolution MRI imaging of implanted tumors can be obtained by intraperitoneal injection of contrast, and it is consistent with pathologic examinations.

This study reported the obtainment of the full-length cDNA of Salvia miltiorrhiza hairy roots (Abbr: SmHDR, GenBank number: JX233817), via extracting Salvia miltiorrhiza hairy roots total RNA, designing specific primers according to the transcriptome data and using the RACE strategy, and then analyzed it with bioinformatics approaches. On this basis, using the real-time PCR to detect SmHDR gene expression after Ag+ induction, and testing tanshinones contents of corresponding samples by UPLC. SmHDR has 1 647 nucleotides, and an open reading frame (ORF) encoding a protein of 463 amino acid residues. The deduced protein has isoelectric point (pI) of 5.72 and a calculated molecular weight about 51.88 kD. In the secondary structure, the percentage of alpha helix, beta turn and random coil were 35.64%, 20.30% and 44.06%, respectively. Sequence alignment and phylogenetic analysis demonstrated that SmHDR had relative close relationship to the HDR of Picrorhiza kurrooa, similar to HDR from other species of plants. Real time PCR results indicated that elicitor of Ag+ stimulated the increase of mRNA expression of SmHDR. At the same time, results of ultra performance liquid chromatography (UPLC), used to examine the accumulation of diterpenoid tanshinones in hairy roots, showed that the contents of diterpenoid tanshinones in hairy roots of Salvia miltiorrhiza were increased dramatically at 12 h after treated with Ag+, and then decreased significantly. This result showed a positive correlation between the levels of mRNA expression and tanshinones accumulation in Salvia miltiorrhiza stimulated by Ag+. The content of tanshinones was gradually raised, and it had an obvious increase at 120 h. The bioinformatics analysis and gene expression indicated that SmHDR might be involved in tanshinones biosynthesis, which laid the foundation for further study of secondary metabolic regulation mechanism of tanshinones.