[Cloning and expression analysis of CpPMM gene in Codonopsis pilosula].

GDP-mannose is an important precursor for the synthesis of Codonopsis pilosula polysaccharides and involved in the synthesis of sugar chains. Phosphomannomutase(PMM)catalyzes the conversion of mannose-6-phosphate(Man-6-P)to mannose-1-phosphate(Man-1-P)to synthesize GDP-mannose. In this study, specific primers were designed based on the PMM gene sequence information in transcriptome data, and the full length of the C. pilosula PMM gene was cloned and named CpPMM. The correlation between the CpPMM gene expression and C. pilosula polysaccharide synthesis was analyzed by a series of bioinformatics analysis, prokaryotic expression and qRT-PCR. The results show that the CpPMM gene contains a 741 bp open reading frame(ORF), encoding 246 amino acids, which is highly similar to the PMM of other species and highly homologous to the Helianthus annuus from the Asteraceae family. It was predicted to be a hydrophilic non-transmembrane protein without signal peptide, which was predicted to be located in the cytoplasm with multiple phosphorylation sites. Combined with predictive analysis of conserved domains, this protein belongs to the HAD(haloacid dehalogenase)superfamily; prokaryotic expression studies show that the size of the CpPMM fusion protein is about 29 kDa, which is consistent with the relative molecular mass predicted. The target protein is an inclusion body and is partially soluble. The qRT-PCR results showed that the CpPMM gene exerted spatiotemporal expression patterns, and the expression level in fruiting period was significantly higher than that in the other three periods such as the flowering period. Along with the growth period of C. pilosula, the polysaccharide content of C. pilosula showed a gradual increase trend, reaching the highest during the harvest time. And there are significant differences in the polysaccharide content of C. pilosula in each period. In this study, the CpPMM gene was cloned from the root of C. pilosula, at the same time, the prokaryotic expression system was constructed. In addition, its gene expression level is highly correlated with the polysaccharide content of C. pilosula. It lays the foundation for further studying the function of CpPMM gene and the analysis of biosynthetic pathways of polysaccharides in medicinal plants.

Study on HPLC specific chromatograms of Lu Dangshen / 中国中药杂志

In this paper, the RP-HPLC specific chromatography was adopted, with DIKMA-C18 (4.6 mm x 250 mm, 5 µm) as the chromatographic column, with a gradient elution compose of acetonitrile and 0.1% phosphoric acid at flow rate of 0.8 mL · min(-1), the detection wavelength was 220 nm. The difference of the HPLC specific chromatograms between the Lu Dangshen and other different base sources and different producing area of Codonopsis Radix was compared, involved in the similarities and differences of the number and the relative peak area of characteristic peaks in the HPLC specific chromatograms. The HPLC specific chromatograms of Lu Dangshen was established and the relative retention times of seven peaks was determined, and the peaks of codonopyrrolidium B, syringin, lobetyolin, tangshenoside I and atractylenoide III were identified; The HPLC specific chromatograms of Lu Dangshen provided a method for scientific evaluation and effective control the quality of Lu Dangshen from Shanxi famous-region.

Peptides analysis in digested edible bird's nest by HPLC-MS / 中国中药杂志

Edible bird's nest contains lots of glycoproteins. The glycosylation inhomogeneity for glycoprotein often results in wide range of molecular weight and the difficulty for protein separation and charaterization. In this paper, proteins in the edible bird's nest were extracted using multiple extractions, and then digested by PNgase F and trypsin. The digest mixture was separated with HPLC, and peptides were identified based on MS/MS data searching. The results indicated that the extracted proteins were amount to 79.7% of total protein in the edible bird's nest. More than 20 species of peptides in the digested mixture were identified. The sequences of these peptides showed similarity with some proteins from Swiss-prot. The research indicated that deglycosylation, tryptic digestion coupled with HPLC-MS/MS is a proper strategy for characterization of proteins in the edible bird's nest.

Calculus formation in the prostatic cavity after transurethral resection of the prostate: causes, treatment and prevention / 中华男科学杂志

<p><b>OBJECTIVE</b>To study the causes, clinical manifestations, treatment and prevention of calculus that develops in the prostatic cavity after transurethral resection of the prostate.</p><p><b>METHODS</b>We reported 11 cases of calculus that developed in the prostatic cavity after transurethral resection or transurethral plasmakinetic resection of prostate. The patients complained of repeated symptoms of frequent micturition, urgent micturition and urodynia after operation, accompanied with urinary tract infection and some with urinary obstruction, which failed to respond to anti-infective therapies. Cystoscopy revealed calculi in the prostatic cavity, with eschar, sphacelus, uneven wound surface and small diverticula in some cases. After diagnosis, 1 case was treated by holmium laser lithotripsy and a second transurethral resection of the prostate, while the other 10 had the calculi removed under the cystoscope, followed by 1 -2 weeks of anti-infective therapy.</p><p><b>RESULTS</b>After treatment, all the 11 cases showed normal results of routine urinalysis, and no more symptoms of frequent micturition, urgent micturition and urodynia. Three- to six-month follow-up found no bladder irritation symptoms and urinary tract infection.</p><p><b>CONCLUSION</b>Repeated symptoms of frequent micturition, urgent micturition, urodynia and urinary tract infection after transurethral resection of the prostate should be considered as the indicators of calculus in the prostatic cavity, which can be confirmed by cystoscopy. It can be treated by lithotripsy or removal of the calculus under the cystoscope, or even a second transurethral resection of the prostate. For its prevention, excessive electric coagulation and uneven wound surface should be avoided and anti-infection treatment is needed.</p>

Effects of hyperbaric oxygen on tumor growth in the mouse model of LNCaP prostate cancer cell line / 中华男科学杂志

<p><b>OBJECTIVE</b>To assess the safety of hyperbaric oxygen in the treatment of radiation-induced hemorrhagic cystitis in patients with prostate cancer, and to investigate its effect on the growth of indolent prostate cancer in vivo.</p><p><b>METHODS</b>Thirty severe combined-immunodeficient mice received subcutaneous injection of human prostate cancer LNCaP cells. Then they were randomized to an experimental and a control group and exposed to 20 sessions of hyperbaric oxygen and normobaric air, respectively, followed by a 4-week observation on the growth of the transplanted tumors and analyses of their histopathological features at 28 days, including the volume, microvessel density (CD34), apoptosis markers (p53 and p27 proteins) and the proliferation index (Ki-67) of the LNCaP tumors.</p><p><b>RESULTS</b>On the 28th day after tumor vaccination, the tumor volume was (120 +/- 7.9) mm3 in the HBO and (122 +/- 8.2) mm3 in the control group; the microvessel density and the expressions of Ki-67, p53 and p27 were 39.3 +/- 5.2, (78.1 +/- 7.6)%, (40.4 +/- 6.2)% and (63.7 +/- 5.1)% in the former, and 36.2 +/- 4.9, (75.3 +/- 8.4)%, (44.2 +/- 5.7)% and (61.5 +/- 5.5)% in the latter. There were no significant differences in all the indexes above between the two groups (P > 0.05).</p><p><b>CONCLUSION</b>Hyperbaric oxygen did not promote the growth of indolent prostate cancer in the murine model, nor did it have any significant effect on the new vessels.</p>

Simultaneous determination of baicalin, rhein and berberine in rat plasma by column-switching high-performance liquid chromatography.

A simple HPLC method using column-switching and ultraviolet detection was developed for the simultaneous determination of baicalin (BA), rhein (RH) and berberine (BE) in rat plasma. Plasma samples were injected directly onto a Capcell Pak MF C(8) column (150 mm x 4.6 mm i.d.) to remove protein and to be pre-separated by an isocratic elution using 50 mmol/L phosphate sodium (pH 6.85)-acetonitrile (10:1, v/v). After the drug-containing fractions were transferred to a Kromasil C(18) column (150 mm x 4.6 mm i.d.) by a valve switching step, the valve position was switched back and the main separation was performed by an isocratic elution using triethylamine adjusted 20 mmol/L phosphoric acid (pH 6.78)-acetonitrile (4:1, v/v). The flow rate was always 1.0 mL/min. The calibration curve showed excellent linear relationship (r>or=0.9997) over the concentration range of 0.4-7.9 microg/mL for baicalin, 0.2-7.8 microg/mL for rhein and 0.4-7.7 microg/mL for berberine in rat plasma. The intra- and inter-day assay precisions (R.S.D.) of three analytes were in the range of 0.34-4.3% and the accuracies were between 98.0% and 102.4%. Their recoveries were all greater than 95%. The method was successfully applied to the multi-constituents plasma concentration-time curve study after oral administration of a traditional Chinese medicine prescription Xiexin-Tang in rats.

Value of preoperative urodynamics to the prognosis of transurethral prostate resection for benign prostatic hyperplasia / 中华男科学杂志

<p><b>OBJECTIVE</b>To establish the value of preoperative urodynamics to the outcome prediction of transurethral prostate resection (TURP) for benign prostatic hyperplasia (BPH).</p><p><b>METHODS</b>One hundred and sixty patients with BPH underwent urodynamic tests. Preoperative IPSS (International Prostate Symptom Score) and QOL (Quality of Life) were compared with those 8 to approximately 11 months after operation.</p><p><b>RESULTS</b>The parameters of urodynamic tests (max. free flow, detrusor pressure at max. flow, Schafer grade, Abrams-Griffiths No, urethral resistance factor, cystometric capacity, effective capacity). IPSS and QOL were improved after operation (P < 0.001). And all the relative coefficients of linear dependence analysis, IPSS, QOL and max. free flow, detrusor pressure at max. flow, Schafer grade, Abrams-Griffiths value, urethral resistance factor, cystometric capacity, and effective capacity conducted postoperatively, were statistically significant.</p><p><b>CONCLUSION</b>Preoperative urodynamics of transurethral prostate resection for benign prostatic hyperplasia may provide indication for operation and predict postoperative improvement of symptoms. Preoperative urodynamics should be considered as a routine examination.</p>

Determination of lignans of Schisandra medicinal plants by HPLC / 中国中药杂志

<p><b>OBJECTIVE</b>To determinate the contents of lignans in the stems, roots and fruits of Schisandra medicinal plants.</p><p><b>METHOD</b>HPLC was applied to determine the contents of schisandrin, gomisin A, schisantherin A, deoxyschizandrin, d-epigalbacine, (+)-anwulignan, wuweizisu B, 6-O-benzoylgomisin O and wuweizisu C in the stems, roots and fruits of Schisandra.</p><p><b>RESULT</b>The percent contents of 9 lignans in the fruits of 5 species were 0.52% - 1.96%, and 0.02% - 1.51% in the stems of 11 species. The lignans in the fruits of S. micrantha were similar to that of S. sphenanthera, especially with high content of deoxyschizandrin. The lignans of S. viridis were similar to that of S. chinensis and with high content of schisandrin, gomisin A and wuweizisu B. (+)-Anwulignan was present in the fruits of S. henryi and S. sphenanthera, in the stems of S. micrantha and S. pubescens var. pubinervis, and in the roots of S. bicolor with percentages of 0.77%, 0.42%, 0.50%, 0.26% and 0.38% respectively. d-Epigalbacine was present in the stems of S. pubescens var. pubinervis, S. pubescens and S. glaucescens with percentages of 0.89%, 1.51% and 0.17% respectively.</p><p><b>CONCLUSION</b>The lignans contents in the fruits of Schisandra were higher than that in the roots and stems. The fruits of S. henryi and S. sphenanthera, the stems of S. micrantha and S. pubescens var. pubinervis and the roots of S. bicolor, the stems of S. pubescens var. pubinervis, S. pubescens and S. glaucescens may be served as the resources plants of (+)-anwulignan and d-epigalbacine respectively.</p>

Ribosomal DNA ITS sequences analysis of the Chinese crude drug fructus schisandrae sphenantherae and fruts of Schisandra viridis / 中国中药杂志

<p><b>OBJECTIVE</b>To find the patterns of the rDNA ITS sequence variation of Schisandra sphenanthera and S. viridis, and to establish the molecular biological method for the identification of Fructus Schisandrae Sphenantherae and the fruits of S. viridis.</p><p><b>METHOD</b>PCR products were sequenced directly and the sequences were analyzed with PAUP 4.0b10. NJ systematic tree was obtained with neighbor-joining method.</p><p><b>RESULT</b>The Complete ITS sequence of S. sphenanthera was 691-692 bp, of which there were 282 bp of ITS1 and 246-247 bp of ITS2. The complete sequence of S. viridis was 694-695 bp, consisting of 285-286 bp of ITS1 and 246-247 bp of ITS2. There were three informative sites in ITS1 regions for the two species. In the NJ tree with Kadsura anamosma and K. coccinea as outgroups, five different populations of S. viridis were the monophyletic group with the bootstrap value of 68%. These populations included one from Tianmushan, Zhejiang province, three populations from Jigongshan, Henan Province and the other two populations of S. viridis cited the sequences from GeneBank (registration numbers are AF263438 and AF163703 respectively).</p><p><b>CONCLUSION</b>The rDNA internal transcribed spacer is a good marker to distinguish the Fructus Schisandrae Sphenantherae from the fruits of S. viridis.</p>