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Purpose@#We compared the clinical outcomes of modified procedures for associating liver partition and portal vein ligation for staged hepatectomy (ALPPS) based on a risk-reduced strategy with those of classic ALPPS procedures in treating large liver carcinoma. @*Materials and Methods@#Short-term outcomes, increases in future liver remnant (FLR) and functional FLR (FFLR), and overall survival (OS) were compared between 45 consecutive patients treated with modified ALPPS procedures and 34 patients treated with classic ALPPS procedures. @*Results@#Clinical outcomes after the 1st-stage operation markedly improved with the modified procedures. Although the proportions of liver cirrhosis and hepatocellular carcinoma were higher in the modified group, the mortality and incidence of severe complications did not increase. FLR and FFLR hypertrophy at 1 week after the 1st-stage operation were similar in both groups; however, kinetic growth rates in the modified group were lower. OS rates were similar. @*Conclusion@#Modified ALPPS procedures could be safely applied to provide long-term survival for patients with liver cirrhosis without sufficient FLR.
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Purpose@#We compared the clinical outcomes of modified procedures for associating liver partition and portal vein ligation for staged hepatectomy (ALPPS) based on a risk-reduced strategy with those of classic ALPPS procedures in treating large liver carcinoma. @*Materials and Methods@#Short-term outcomes, increases in future liver remnant (FLR) and functional FLR (FFLR), and overall survival (OS) were compared between 45 consecutive patients treated with modified ALPPS procedures and 34 patients treated with classic ALPPS procedures. @*Results@#Clinical outcomes after the 1st-stage operation markedly improved with the modified procedures. Although the proportions of liver cirrhosis and hepatocellular carcinoma were higher in the modified group, the mortality and incidence of severe complications did not increase. FLR and FFLR hypertrophy at 1 week after the 1st-stage operation were similar in both groups; however, kinetic growth rates in the modified group were lower. OS rates were similar. @*Conclusion@#Modified ALPPS procedures could be safely applied to provide long-term survival for patients with liver cirrhosis without sufficient FLR.
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This study aims to explore the relationship of DNA methylation with the contents of the index components as well as the growth and development of Pogostemon cablin. The demethylation reagent 5-azacytidine(5-azaC) was used to treat the tissue culture seedlings of patchouliol-type P. cablin. High performance liquid chromatography was employed to evaluate the changes of DNA methy-lation in P. cablin, and GC-MS to detect the contents of index components in P.cablin. The agronomic characters of P.cablin were measured using the common methods. The results showcased that DNA methylation of P.cablin was significantly reduced by 5-azaC in a concentration-dependent manner. Thirty days after treatment with 5-azaC at different concentrations, the content of patchouli alcohol changed slightly; compared with that in the control group, the content of pogostone in 50 μmol·L~(-1) and 100 μmol·L~(-1) 5-azaC groups was significantly up-regulated. The 100 μmol·L~(-1) 5-azaC group had the largest differences in contents of pogostone and patchouli alcohol compared with the control group, followed by the 50 μmol·L~(-1) 5-azaC group. Ninety days after disinhibition, the content of pogostone in the treatment group was significantly increased and the content of patchouli alcohol was significantly decreased. In addition, 5-azaC significantly inhibited the growth and development of P.cablin in a dose-dependent manner. These results indicate that DNA methylation regulates the biosynthesis of the index components in patchouliol-type P.cablin and proper demethylation can directly promote the synthesis of pogostone and indirectly affect the accumulation of patchouli alcohol.
Subject(s)
Azacitidine , DNA Methylation , Gas Chromatography-Mass Spectrometry , Oils, Volatile , Pogostemon/geneticsABSTRACT
The terpenoids in Pogostemon cablin have complex structures and abundant pharmacological effects. Patchouli alcohol(PA) and pogostone(PO) have a high medicinal value by virtue of anti-tumor, anti-inflammatory, antibacterial, antioxidant, and other biological activities. Due to the low content of terpenoid metabolites in P. cablin, the study of biosynthesis and metabolism regulation can provide a biosynthetic basis for obtaining high-content terpenoids. In this study, key enzyme genes in biosynthesis, transcription factors in metabolism regulation, spatio-temporal expression of terpene synthase were reviewed, aiming to provide a reference for the development, protection, and utilization of P. cablin resources.
Subject(s)
Pogostemon/genetics , Terpenes , Transcription Factors/geneticsABSTRACT
Objective:This paper aims to study the genetic diversity of <italic>Pogostemon cablin</italic> by amplified fragment length polymorphism （AFLP） markers. Method:The 12 pairs of primers were used for AFLP analysis of 212 samples from 14 varieties，and biological analysis software such as POPGENE 32，Arlequinver 3.5，MEGA 7 and NTSYSpc 2.10e were used for polymorphism parameter calculation，principal coordinate analysis and cluster analysis. Result:A total of 2 238 loci were amplified by 12 pairs of primers. 2 226 of them were polymorphic loci， accounting for 99.38%. At the inter-population level，the values of effective alleles（<italic>Ne</italic>），Nei's gene diversity index（<italic>H</italic>），Shannon polymorphic information index（<italic>I</italic>） were 1.365 6±0.066 3， 0.220 7±0.036 4， and 0.343 7±0.050 2，respectively；and 1.118 5±0.038 7，0.071 3±0.023 0，0.109 4±0.035 0，respectively at the intra-population level. Analysis of molecular variance（AMOVA）showed that 71.57% of the total variation of <italic>P. cablin</italic> was of inter-population nature， and 28.43% was of intra-population nature. The 14 populations could be divided into four groups by cluster analysis. Conclusion:The results of AFLP molecular markers showed that abundant genetic diversity was present at inter-population level of <italic>P. cablin</italic>，however，relatively low at intra-population level； the genetic differentiation at the inter-population level was significant，which could provide a reference for the subsequent study of good germplasm selection of <italic>P. cablin</italic>.
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Objective: The molecular identification method of Polygonum multiflorum from different producing areas was explored by using the sequencing of intergenic region of chloroplast genes psbA-trnH. Methods: A total of 116 samples of P. multiflorum were collected from 15 populations in seven provinces and autonomous regions. The total DNA was extracted and the sequences of psbA-trnH were amplified by PCR. The purified PCR products were sequenced and analyzed by MEGA 6.06 software. Results: The genetic distances among the populations of P.multiflorum are 0.001—0.187. In the maximum likelihood phylogenetic tree, 15 populations of P. multiflorum were clustered into two blanches. Conclusion: The genetic variation of P. multiflorum is significant and the psbA-trnH sequences of P. multiflorum can be used as germplasm source for molecular identification between Deqing regarded as geo-authentic habitat and other producing areas.
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OBJECTIVE: To establish a genebank for phage single-chain antibody for further screening the specificity of single chain fragment variable( Sc Fv) in lung tissue of silicosis rats by phage display technology. METHODS: Twenty-four specific pathogen free male SD rats were used to construct silicosis model by one-time bronchial perfusion with 1. 0 m L of silicon dioxide suspension( mass concentration,100 g / L). We took periphery blood from 6 rats 3,6,9 and 12 weeks respectively after establishing the model. The peripheral lymphocytes were mixed,and total RNA was extracted using Trizol,and c DNA was synthesized by reverse transcription. The degenerated primers were used to amplify the variable region of heavy chain( VH) gene and variable region of light chain( VL) gene by polymerase chain reaction( PCR).Then VH and VL genes were assembled to form Sc Fv by T4 DNA linker. The cloning recombinant of Sc Fv and plasmid of PCANTAB-5e were transformed into competence E. coli TG1 by calcium chloride. The Sc Fv genebank of silicosis model was constructed by M13K07 helper phage superinfection. There were 10 bacterial colonies for plasmid restriction dualenzyme digestion randomly selected for confirmation. RESULTS: Agarose gel electrophoresis showed that there were two bands of obvious 28 S and 18 S in total RNA of periphery blood lymphocytes of silicosis rats. The total RNA was intact. The size of VH gene fragment was about 400 bp,the size of VL gene fragment was about 350 bp and recombinant Sc Fv gene fragment length was about 750 bp. The helper phage was amplified and placed with double-deck agar plate and observed limpid plaque with the size of a rice grain. The phage titer was 1. 35 × 10~(16) pfu / L. The recombinant plasmids were transformed into E. coli TG1 and total bacterial count was 8. 0 × 10~9 cfu / L in resistant plate. The positive cloned plasmid PCR gel electrophoresis and double enzyme results showed a positive inserting rate of 90. 0%. The capacity of phage single-chain antibody genebank of experimental silicosis was 7. 2 × 10~9 cfu / L. CONCLUSION: The silicosis rat model with phage Sc Fv gnebank could be successfully established,and its capacity and diversity provide support for the follow-up screening.
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<p><b>OBJECTIVE</b>The purpose of this review is to discuss some critical issues of isoflavones protective against the development of prostate cancer (PCa).</p><p><b>DATA SOURCES</b>Data cited in this review were obtained primarily from PubMed and Embase from 1975 to 2015.</p><p><b>STUDY SELECTION</b>Articles were selected with the search terms "isoflavone", "Phytoestrogen", "soy", "genistin", and "PCa ".</p><p><b>RESULTS</b>Isoflavones do not play an important role on prostate-specific antigen levels reduction in PCa patients or healthy men. The effect of isoflavones on sex hormone levels and PCa risk may be determined by equol converting bacteria in the intestine, specific polymorphic variation and concentrations of isoflavones. The intake of various types of phytoestrogens with lower concentrations in the daily diet may produce synergistic effects against PCa. Moreover, prostate tissue may concentrate isoflavones to potentially anti-carcinogenic levels. In addition, it is noteworthy that isoflavones may act as an agonist in PCa.</p><p><b>CONCLUSIONS</b>Isoflavones play a protective role against the development of PCa. However, careful consideration should be given when isoflavones are used in the prevention and treatment of PCa.</p>
Subject(s)
Humans , Male , Isoflavones , Therapeutic Uses , Phytoestrogens , Therapeutic Uses , Prostatic NeoplasmsABSTRACT
Objective To explore the effect of colectomy on the expression of glucagon-like peptide-2 (GLP-2) in serum and ileum and compare the changes of GLP-2 after different colectomy in rats.Methods Eighty male or female rats,aging 3-4 months old,were recruited in this study.The rats weight 180-250 g.The 80 rats were evenly and randomly distributed into 4 groups according to the surgical procedures they underwent:control group,in which the rats were not performed any procedures; sham surgery group,in which the rats underwent laparotomy ;left hemicolectomy group,in which the rats were performed left hemicolectomy;and subtotal colectomy group,in which the rats were performed subtotal colectomy.According to execution time,each group had 4 subgroups,including 0 day group,10 day group,20 day group and 30 day group,and 5 rats were included in each subgroup.The whole blood was collected through cardiac puncture.Serum was collected and GLP-2 in serum was determined by enzyme-linked immunosorbent assay.The GLP-2 in ileum was studied by immunohistochemistry staining.The length of villus of ileum was also measured by HE staining.Results The protein of GLP-2 was found in fibroblasts,epithelium and endocrine cells in ileum.Compared with control group and sham surgery group,the expressions of GLP-2 in serum and ileum increased significantly in the rats that underwent left hemicolectomy after surgery(all P ＜ 0.05),and it was increasing with time.The villus length of the rats underwent left hemicolectomy also increased significantly compared with the control group and sham surgery groups 20 days and 30 days after surgery(all P ＜ 0.05).The expressions of GLP-2 in serun and ileum and villus length of the rats with subtotal colectomy were not significantly different from those of the control group and sham surgery group(all P ＞ 0.05).Conclusions The expressions of GLP-2 in serum and ileum are elevated after left hemicolectomy but not in subtotal colectomy,which may be related to the delayed intestinal adaption after colectomy.
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<p><b>OBJECTIVE</b>Based on the observation that coagulation necrosis occurs in the majority of neonatal necrotizing enterocolitis (NEC) patients, it is clear that intestinal ischemia is a contributing factor to the pathogenesis of NEC. However, the published studies regarding the role of intestinal ischemia in NEC are controversial. The aim of this paper is to review the current studies regarding intestinal microcirculatory dysfunction and NEC, and try to elucidate the exact role of intestinal microcirculatory dysfunction in NEC.</p><p><b>DATA SOURCES</b>The studies cited in this review were mainly obtained from articles listed in Medline and PubMed. The search terms used were "intestinal microcirculatory dysfunction" and "neonatal necrotizing enterocolitis".</p><p><b>STUDY SELECTION</b>Mainly original milestone articles and critical reviews written by major pioneer investigators in the field were selected.</p><p><b>RESULTS</b>Immature regulatory control of mesentery circulation makes the neonatal intestinal microvasculature vulnerable. When neonates are subjected to stress, endothelial cell dysfunction occurs and results in vasoconstriction of arterioles, inflammatory cell infiltration and activation in venules, and endothelial barrier disruption in capillaries. The compromised vasculature increases circulation resistance and therefore decreases intestinal perfusion, and may eventually progress to intestinal necrosis.</p><p><b>CONCLUSION</b>Intestinal ischemia plays an important role through the whole course of NEC. New therapeutic agents targeting intestinal ischemia, like HB-EGF, are promising therapeutic agents for the treatment of NEC.</p>
Subject(s)
Humans , Infant, Newborn , Endothelin-1 , Physiology , Endothelium, Vascular , Enterocolitis, Necrotizing , Drug Therapy , Pathology , Heparin-binding EGF-like Growth Factor , Intercellular Signaling Peptides and Proteins , Therapeutic Uses , Intestines , Ischemia , Microcirculation , Physiology , Nitric Oxide , Physiology , Splanchnic CirculationABSTRACT
To study the effects of Smad4 on liver fibrosis and hepatocarcinogenesis in mice treated with CCl(4)/ethanol. The wild-type mice (Smad4 +/+) and the Smad4 knockout mice (Smad4 +/-) were injected subcutaneously with carbon tetrachloride(CCl(4))/ethanol twice a week for twenty weeks. The expression of Smad4, TGFbeta1, Smad2, Smad3, Smad6, TIMP1, MMP2 and MMP9 was detected by RT-PCR. In the cirrhotic liver, the expression of Smad4 mRNA was significantly higher than that in the normal liver. Comparing with wild-type mice (Smad4 +/+), the TGFbeta1-Smad4 signaling was markedly attenuated in the Smad4 knockout mice (Smad4 +/-). After induction by CCl(4)/ethanol, the hepatic fibrosis in the Smad4 knockout mice (Smad4 +/-) was obviously alleviated compared with the wild-type mice (Smad4 +/+), and the incidence rate of hepatocarcinogenesis of the former was also lower than that of the latter(32.0% vs 41.9%). These results indicate that knocking out Smad4 can delay the progression of liver fibrosis and liver cancer.
Subject(s)
Animals , Female , Male , Mice , Carbon Tetrachloride , Disease Models, Animal , Ethanol , Liver Cirrhosis, Experimental , Metabolism , Pathology , Liver Neoplasms, Experimental , Metabolism , Pathology , Mice, Knockout , RNA, Messenger , Genetics , Metabolism , Reverse Transcriptase Polymerase Chain Reaction , Signal Transduction , Smad Proteins , Genetics , Metabolism , Smad4 Protein , Genetics , Metabolism , Tissue Inhibitor of Metalloproteinase-1 , Genetics , Metabolism , Transforming Growth Factor beta1 , Genetics , MetabolismABSTRACT
<p><b>OBJECTIVE</b>to compare the therapeutic effect of posterolateral fusion (PLF), posterior lumbar interbody fusion (PLIF) and posterior circumferential fusion (PCF) for lumbar spondylolisthesis.</p><p><b>METHODS</b>from January 2003 to December 2008, 232 patients with lumbar spondylolisthesis treated with pedicle screw fixation and followed for reviewed retrospectively. The patients were divided into three groups based on fusion method: group A (PLF, 66 case), group B (PLIF, 54 case)and group C (PCF, 112 case). The three groups were reviewed and compared for clinical outcome and fusion rate.</p><p><b>RESULTS</b>the mean follow-up period was 21 months (range, 6-60 months). The fusion rate was 80.1% for PLF, 92.5% for PLIF and 93.7% for PCF group (P > 0.05). As to isthmic spondylolisthesis or Meyerding grade degenerative II and III spondylolisthesis, the fusion rate was 60.7% for PLF group, 90% for PLIF group and 93.3% for PCF group (P < 0.05). Compare the fusion rate for PLF group and PLIF+ PCF group (P < 0.05), fusion rate for PLIF group and PCF group (P > 0.05). The rate of excellent and good together was 84.8% in PLF group, 90.7% in PLIF group and 93.6% in PCF group (P > 0.05).</p><p><b>CONCLUSIONS</b>posterior lumbar interbody fusion and posterior circumferential fusion are more consistent with bio-mechanics, have a higher fusion rate, for the treatment of spondylolisthesis they are the preferred surgical approaches.</p>
Subject(s)
Adolescent , Adult , Aged , Female , Humans , Male , Middle Aged , Young Adult , Bone Transplantation , Methods , Follow-Up Studies , Lumbar Vertebrae , General Surgery , Retrospective Studies , Spinal Fusion , Methods , Spondylolisthesis , General Surgery , Treatment OutcomeABSTRACT
Objective: To develop a high efficient expression, purification system of recombinant arginine deiminase(ADI).Methods: cDNA fragment encoding for mycoplasma ADI was obtained by artificial synthesis and was cloned into prokaryotic expression vector(pBV220). The recombinant ADI was generated by the transformation of the recombinant vector into the host strain DH5?. Anion exchange and gel filtration chromatography was carried out for purification of the recombinant ADI. The biological activity of final product was detected by the assay of agrinine degradation in vitro. Results: A prokaryotic expression plasmid pBV220-ADI was generated successfully, and was identified by DNA sequencing; the recombinant protein was highly expressed in DH5?, the proportion of the recombinant protein is exceeded 35% of the whole protein. The inclusion bodies were solubilized with 6mol/L guanidine hydrochloride under reducing conditions in order to avoid incorrect disulfide-bond formation of the recombinant ADI molecules. Dilution and dialysis at lower degrees temperature were the optimum renaturation methods. After gel filtration, the purity and specific activity of rADI reached 95% and 80 IU/mg respectively. Conclusions: A set of protocols for high efficient rADI expression and purification has been established, which is simple, efficient and applicable.
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<p><b>BACKGROUND</b>Rheumatic heart disease (RHD) is the most important sequela of rheumatic fever (RF): evidence that streptococcal infection is aetiological is prominent, but sometimes contradictory. Acute HSV-1 infection in mouse leads to carditis and valvulitis whereas recurrent infection results in inflammatory granulomatous lesions that resemble Aschoff bodies. Cells containing HSV-1 inclusions or virus infected giant cells appear similar to Anitschkow cells or Aschoff cells respectively. We hypothesized that HSV-1 infection also may be involved in RHD.</p><p><b>METHODS</b>Formalin-fixed, paraffin-embedded valvular tissue samples from 32 patients with RHD were investigated for evidence of HSV-1 infection. HSV-1 antigen was detected by immunohistochemistry, using HSV-1-specific monoclonal and polyclonal antibodies. HSV-1 glycoprotein D gene sequences were amplified by nPCR, using beta-globin gene amplification in the same samples as internal control. Valvular tissue from 5 cases of sudden death and 3 cases died of neisseria meningitis without a history of valvular disease was used for comparison. HSV-1-infected lung tissue was used as positive control.</p><p><b>RESULTS</b>HSV-1 antigens were detected in valvular tissues from 21 of 32 (65.6%) patients. Fifteen of these 21 (46.9% of cases), but no antigen-negative sample, were positive also for HSV DNA. Nucleotide sequence of PCR products was homologous to the targeted region of the HSV-1 glycoprotein D gene. HSV-1 antigen was present also in one case of sudden death but viral DNA was not found in any tissue sample from the comparison group. Results from reagent and positive controls were as anticipated.</p><p><b>CONCLUSIONS</b>This is the first study to show the presence of HSV-1 antigen and genomic DNA in valvular tissues from patients with RHD and provides evidence for an association of HSV-1 infection with some cases of rheumatic valvular disease.</p>