An actinomycete B37 was isolated from an intertidal marine sponge Hymeniacidon perleve, which has strong activity against Gram positive bacteria and moderate activity against tumor cells. The mycelium and spore morphology, physiological properties and 16SrDNA sequence suggested that B37 is Streptomyces pseudogriseolus. The fermentation conditions of this strain were investigated for the biosynthesis of bioactive metabolites.

This study is to determine the content of three alkaloids and establish the HPLC fingerprint of "Jianlian" Nelumbinis Plumula. The HPLC method of content determination was as follows: Thermo C18 (4. 6 mm x 250 mm, 5 μm) was conducted with acetonitrile-sodium dodecyl sulfonate solution-acetic acid (56: 43: 1) at a flow rate of 1.0 mL x min(-1). The monitoring wavelength was set at 282 nm and the column temperature was 35 degrees C. The method of HPLC fingerprint was as follows: Agilent ZORBAX SB-Aq C18 (4.6 mm x 250 mm, 5 μm) was conducted with gradient elution of methanol and water at a flow rate of 0.8 mL x min(-1), the monitoring wavelength was set at 282 nm and the column temperature was 35 degrees C. Similarities evaluation and hierarchical clustering analysis were applied to demonstrate the variability of 12 batches of "Jianlian" Nelumbinis Plumula samples. The results demonstrated that 11 batches showed good similarity on chemical constituents. The method could well display the chemical information of "Jianlian" Nelumbinis Plumula. It was simple, reliable and could be used for the chemical quality control of "Jianlian" Nelumbinis Plumula.
Alkaloids ; analysis ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; Nelumbo ; chemistry ; Quality Control ; Seeds ; chemistry

OBJECTIVETo explore the mechanism of electroacupuncture for treating focal cerebral ischemia-reperfusion injury.

METHODSSeventy-five Wistar rats were randomly divided into a control group, a model group and an electroacupuncture group, 25 cases in each group. The model of focal cerebral ischemia-reperfusion was established by inserting nylon thread into the internal carotid artery except the control group which was only separated of the carotid artery without occlusion. Electroacupuncture group was treated with electroacupuncture at "Baihui (GV 20)" and "Dazhui (GV 14)" and the other groups without electroacupuncture treatment. The number of nestin positive cells expression at 1st, 3rd, 7th, 14th and 21st days after focal cerebral ischemia reperfusion was observed by use of immunohistochemistry method.

RESULTSThe number of nestin positive cells in electroacupuncture group at ischemia side was significantly more than that in the model group at 3rd, 7th, 14th and 21st days (P < 0.05, P < 0.01), and at contralateral ischemia side, the number of nestin positive cells in the electroacupuncture group was significantly more than that in the model group only at 7th day (P < 0.05).

CONCLUSIONElectroacupuncture at "Baihui (GV 20)" and "Dazhui (GV 14)" in rats can increase the number of nestin positive cells in hippocampus after focal cerebral ischemia-reperfusion, which may be one of the important mechanisms of electroacupuncture in treating acute cerebral ischemic diseases.

Animals ; Brain Ischemia ; genetics ; metabolism ; surgery ; therapy ; Disease Models, Animal ; Electroacupuncture ; Gene Expression ; Hippocampus ; cytology ; metabolism ; Humans ; Intermediate Filament Proteins ; genetics ; metabolism ; Male ; Nerve Tissue Proteins ; genetics ; metabolism ; Nestin ; Neural Stem Cells ; metabolism ; Random Allocation ; Rats ; Rats, Wistar ; Reperfusion

OBJECTIVETo compare the outcomes of the three microsurgical strategies, inguinal high ligation (IHL), retroperitoneal high ligation (RHL) and low ligation (LL) of internal spermatic veins, in the treatment of varicocele.

METHODSWe retrospectively analyzed 120 cases of varicocele, which were equally divided into groups I , II and III to be treated by IHL, RHL and LL of internal spermatic veins, respectively. We compared the operation times, post-operative complications, recurrence rates and results of pre- and post-operation semen analysis among the three groups.

RESULTSThe mean operation time was significantly longer in group III ( [55 +/- 6 ] min) than in I ([35 +/- 10] min) and II ([42 +/- 12] min) (P<0.05), while the rate of post-operative complications remarkably higher in group I (4 cases of hydrocele [10% ] and 3 cases of epididymitis [7.5%]) than in II (2 cases of hydrocele [5%] and 2 cases of epididymitis [5%]) and III (1 case of hydrocele [2.5%] and 1 case of epididymitis [2.5%]) (P<0.05). Six to 12 (mean 9) months of follow-up visit found 2 cases (5% ) of recurrence in group I, 1 case (2.5%) in group II and none in group III, with no statistically significant difference among the three groups (P>0.05). At 12 months after surgery, group III showed significantly higher sperm concentration, grade a +b sperm and the sperm motility than the other two (P<0.05), but no significant differences were observed in these parameters among the three groups preoperatively.

CONCLUSIONAs a microsurgical approach to the treatment of varicocele, LL is better than IHL and RHL of internal spermatic veins in improving the seminal parameters of the patients.

Humans ; Ligation ; Male ; Microsurgery ; adverse effects ; methods ; Retrospective Studies ; Treatment Outcome ; Urologic Surgical Procedures, Male ; adverse effects ; methods ; Varicocele ; surgery

OBJECTIVETo explore the effect of terbutaline on sodium transport in rat alveolar type I (ATI) and type II (ATII) cells of rats.

METHODSThe whole cell currents were recorded from ATII cells isolated from rat lungs perfused with or without amiloride (inhibitor of epithelial sodium channel) and ZnCl(2) (inhibitor of cyclic nucleotide-gated cation channel) in the whole cell recording mode using the patch-clamp technique. The effect of terbutaline on the currents was examined.

RESULTSThe main currents recorded from ATII cells were amiloride-sensitive and Zn(2+)-sensitive. The amiloride-sensitive and Zn(2+)-sensitive current shared a similar proportion (P>0.05). Both currents could be significantly increased by terbutaline (P<0.05), and the proportion of amiloride-sensitive current was 1.7 times that of Zn(2+)-sensitive current (P<0.05).

CONCLUSIONThere are functional epithelial sodium channels (ENaC) and cyclic nucleotide-gated cation channels (CNG) on freshly isolated ATII cells, both serving as the main channels for sodium transport. Terbutaline increases the absorption of alveolar fluid primarily by increasing sodium transport of ENaC and CNG on ATI and AT II cells.

Amiloride ; pharmacology ; Animals ; Chlorides ; pharmacology ; Cyclic Nucleotide-Gated Cation Channels ; antagonists & inhibitors ; drug effects ; Male ; Peptides ; pharmacology ; Pulmonary Alveoli ; cytology ; metabolism ; Rats ; Rats, Sprague-Dawley ; Sodium ; metabolism ; Sodium Channels ; drug effects ; Terbutaline ; pharmacology ; Zinc Compounds ; pharmacology

Major surface protein (p30) and Dense Granule Antigen GRA6 of Toxoplasma gondii have good antigenicity, and could be used for detection of IgM against Toxoplasma gondii. GRA6 may complement P30 to reach more high sensitivity for detection of antibodies to Toxoplasma gondii, so, we try to express the chimeric protein of GRA6 and P30 by genetic engineering, identify its antignenicity and use for developing diagnosis reagent. Antigenic domains of p30 and GRA6 of Toxoplasma gondii were screened by analyzing their sequences using the software ANTHEWIN. Two DNA fragments encoding respectively antigenic domains of p30 and GRA6 were cloned, they were inserted into the same expression vector pET28a( + ) and expressed as a chimeric protein in Escherichia coli. BL21(DE3), the expressed chimeric protein of p30 with GRA6 in a form of inclusion body was about 25% of total proteins of E. coli. BL21(DE3). The inclusion body was washed once with 0.5% Triton X-100 and dissolved with 0.5% SKL, after renaturation by gradient dialysis, the recombinant protein was purified by DEAE-Sepharose FF cation column and then detected with 12% SDS-PAGE, it exists mainly in the eluted peak with 300 mmol/L NaCl and has high purity. By using enzyme-linked immunosorbent assay (ELISA), the recombinant protein was examined for reactivity with immunoglobulin M (IgM) antibodies in 6 sera from patients infected with Toxoplasma gondii ., it was reactive with all the 6 sera but not with sera from normal people, these results showed that the recombinant chimeric antigen has good antigenicity and specificity and could be used for detection of IgM against Toxoplasma gondii. The expressed chimeric protein could be used for epidemic investigation of Toxoplasma gondii, blood donor screening, especially for detection of pregnant women, and is of great significance in prevention of Toxoplasma gondii infection.
Animals ; Antigens, Protozoan ; genetics ; immunology ; metabolism ; Electrophoresis, Polyacrylamide Gel ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Immunoglobulin M ; immunology ; Models, Genetic ; Polymerase Chain Reaction ; Pregnancy ; Protozoan Proteins ; genetics ; metabolism ; Recombinant Fusion Proteins ; genetics ; immunology ; metabolism ; Toxoplasma ; genetics ; immunology

The quality of Chinese medicinal materials relates greatly to the clinical curative effect and security. In order to ensure the quality and safety of Chinese medicinal materials, a systematic and operable traceability system needs to be established. It can realize the whole process of quality and safety management of Chinese medicinal materials "from production to consumption" through recording and inquiring information and recalling defective products, which is an important direction for the future development of traditional Chinese medicine. But it is still at the exploration and trial stage. In this paper, a framework of Chinese medicinal materials' quality and safety traceability system was established on the basis of the domestic and international experience about the construction of food and agricultural products traceability systems. The relationship between traceability system of Chinese medicinal materials' quality and GAP, GMP, GSP was analyzed, and the possible problems and the corresponding solutions were discussed.
China ; Drugs, Chinese Herbal ; chemistry ; standards ; Humans ; Medicine, Chinese Traditional ; standards ; Quality Control

To study the chemical constituents of K. oblongifolia, silica gel column chromatography, MCI and Sephadex LH-20 were used to separate the 70% acetone extract of the stems of K. oblongifolia. The structures of the isolated compounds have been established on the basis of physicochemical and NMR spectroscopic evidence as well as ESI-MS in some cases. Twenty compounds were obtained and identified as heteroclitalignan A (1), kadsulignan F (2), kadoblongifolin C (3), schizanrin F (4), heteroclitalignan C (5), kadsurarin (6), kadsulignan O (7), eburicol (8), meso-dihydroguaiaretic acid (9), kadsufolin A (10), tiegusanin M (11), heteroclitin B (12), (7'S)-parabenzlactone (13), angeloylbinankadsurin B (14), propinquain H (15), quercetin (16), kadsulignan P (17), schizanrin G (18), micrandilactone C (19) and (-)-shikimic acid (20). Compouds 1, 5, 8, 11-15, 18 and 20 were isolated from this plant for the first time. Toxicity of compounds 1-10 were evaluated with zebrafish model to observe the effect on its embryonic development and heart function. The results showed that compounds 7, 9 and 10 caused edema of zebrafish embryo and decreased the heart rate of zebrafish, which exhibited interference effect on heart development of zebrafish.
Animals ; Embryo, Nonmammalian ; drug effects ; Guaiacol ; analogs & derivatives ; toxicity ; Kadsura ; chemistry ; Lignans ; toxicity ; Plant Extracts ; toxicity ; Quercetin ; toxicity ; Triterpenes ; toxicity ; Zebrafish ; embryology

OBJECTIVETo study the validity of transplanting transverse colon to replace esophagus in treating cicatricial stricture resulting from severe esophageal chemical burns in children.

METHODSA retrospective study was carried out on the clinical data of 46 patients with severe chemical esophageal burns who were treated from November 1972 to September 2008. The transverse colon with the ascending branch of the left colic artery was brought through a retrosternal tunnel to replace strictured esophagus. Thirty-two patients underwent colon-esophageal anastomosis and 14 patients underwent colon-pharyngeal anastomosis.

RESULTSAll patients survived after surgery, but complications occurred in 7 cases, including leakage of anastomosis in cervical region in 4 cases, stenosis of anastomosis in 2 cases, and dyspnea in 1 case, and they were cured after due treatment. Follow-up study (1 - 26 years) in 39 patients revealed that there was no difference in growth, development and diet between the patients and the normal children of the same age.

CONCLUSIONSEsophageal reconstruction with transverse colon together with the ascending branch of the left colic artery through a retrosternal tunnel is a valuable method for treating cicatricial stricture of the esophagus secondary to severe chemical burns of the esophagus in children.

Burns, Chemical ; complications ; surgery ; Child ; Child, Preschool ; Cicatrix ; complications ; etiology ; Colon, Transverse ; transplantation ; Esophageal Stenosis ; etiology ; surgery ; Esophagus ; surgery ; Female ; Humans ; Infant ; Male ; Postoperative Complications ; surgery ; Retrospective Studies

Objective:To investigate the effect of downregulation of SLP-2 gene expression on proliferation and apoptosis of brain glioma cells.Methods:The targeting siRNA sequence of SLP-2 transfected U251 human glioma cells,blank group (without any cell treatment)and negative control group (cell transfected nonsense siRNA sequence)were set,Western blot was used to detect the expression of SLP-2,Bcl-2,Bax,Notch1,Hes1 protein after transfected 48 h;cell proliferation was detected by CCKg;cell apoptosis was detected by flow cytometry.Results:The protein expression of SLP-2 in negative control group had no significant difference with the control group (P＞0.05),and the expression of SLP-2 protein in SLP-2 knock down group was significantly lower than control group (P＜0.05);cell survival rate,apoptosis rate,mRNA expression of IL-6,TNF-α and Bcl-2,Bax,Notch1,Hes1 protein expression in the negative control group and the blank group had no significant difference (P＞0.05),cell survival rate,mRNA expression of IL-6 and TNF-α and the expression of Bcl-2,Notch1,Hes1 protein in SLP-2 knock down group was significantly lower than control group,the apoptosis rate and the expression of Bax protein was significantly higher than control group (P＞0.05).Conclusion:Downregulation of SLP-2 gene expression can significantly inhibit the proliferation and induce apoptosis of glioma cells,down regulate the expression of inflammatory cytokines IL-6 and TNF-α,and the mechanism is related to the inhibition of Notch1 signaling pathway.