Objective:To optimize the extraction process of Hegan Lidan granules. Methods:In order to choose the optimal tech-nological parameters, the content of baicalin was determined by HPLC. An orthogonal method was utilized with solvent volume, extrac-tion time and extraction times as the impacting factors and the content of baicalin and extraction rate as the indices. Results:The opti-mal parameters were as follows:using 8-fold water as the solvent, the raw material was extracted three times with 2 h for each. Con-clusion:The process is steady and feasible, and can be used in the extraction of Hegan Lidan granules.

Objective: To establish the quality standard for compound Heishen pills. Methods: Scrophulariae Radix, Radix et Rhizoma and Belamcancae Rhizoma were identified by TLC. HPLC was used to determine the content of harpagoside and cinnamic acid in Scrophulariae Radix on a Welchrom-C18 column using methanol-acetonitrile-1% ethylic acid (8∶21∶71) as the mobile phase. The flow rate was 1. 0 ml·min-1 . The column temperature was at 30℃ and the detection wavelength was set at 278 nm. Results:The TLC method had good specificity without interference from negative control. The linear range of harpagoside was 1. 32-65. 80μg·ml-1 with the average recovery of 98. 06%(RSD=2. 16%),and that of cinnamic acid was 0. 38-19. 20 μg·ml-1 with the average recovery of 98. 78%(RSD=1. 34%). RSDs of precision, stability and reproductibility tests were all below 2%. Conclusion: The established method is accurate, feasible and reproducible. It can be used in the quality control of compound Heishen pills.

Objective To investigate the role of autophagy in radiation-induced death process of human esophageal squamous carcinoma Eca-109 cells.Methods Esophageal carcinoma cell line Eca-109 was divided into 6 groups of control,5 mmol/L 3-Methyladenine treatment,10 mmol/L treatment,6 Gy irradiation,irradiation + 5 mmol/L drug,and irradiation + 10 mmol/L drug.Some cells were transferred with GFP-LC3 plasmid and the changes of autophagosome were obserred.After each treatment,the expression of autophagy marker LC3B was measured by Western Blot,cell viability was detected by MTT,morphological characteristics of apoptosis cells were stained with a fluorescein of Hoechst 33342 and the percentage of apoptotic cells and cell cycle distribution were measured by flow cytometry.Clonogenic survival were used to evaluate the cell radiosensitivity.Results Autophagy level was increased after radiation,and the LC3B Ⅱ expression and LC3B Ⅱ/LC3B Ⅰ ratio were significantly decreased by autophagy inhibitor 3-Methyladenine (F =25.64,P ＜ 0.05).The number of autophagosome fluorescent foci were significantly increased in the GFP-LC3 transfected cells after radiation,but reduced by 3-Methyladenine (F =127.36,P ＜ 0.05).Compared with radiation alone group,autophagy inhibition combined with radiation significantly decreased cell viability (F =129.54,P ＜ 0.05) and colony formation,increased apoptosis and the percentage of G2/M-phase cells.Conclusions 3-Methyladenine enhances the radiosensitivity of esophageal squamous carcinoma Eca-109 cells,suggesting that inhibition of autophagy could be used as an adjuvant treatment of radiotherapy in esophageal squamous carcinoma.

Objective To explore the surgical procedures and effects for recurrent scalp dermatofibrosarcoma protuberans (DFSP).Methods Retrospective review was conducted in 7 scalp recurrent DFSP cases in the past 3 years.Intraoperative frozen section analysis of surgical margins was performed.Wide local excision with margins of 3 cm or more was applied,and excision of the skull external lamina or the entire layer of the cranial bones was performed once the skulls under the tumors were invaded.Frozen section analysis was again performed on all the margins.Secondary wound was repaired with local skin flap or fascial pedicled flaps.Surgically removed tumor was paraffin-embedded for HE-staining and immunohistochemical analysis.Results All of 7 tumor specimens were proved recurrent DFSP with histologic findings of intraoperation or postoperation.7 patients were all performed with wide local excision with margins of 3 cm or more,also with skull excision (5 cases with excision of skull external lamina and 2 cases with excision of the entire layer of skull).Negative margins were confirmed with intraoperative frozen sections or postoperative paraffin-embedded sections analysis were confirmed.Immunohistochemical analysis demonstrated that Ki-67 was all positive and CD34 was partially positive in 7 cases.No tumor recurrence or metastasis had been observed in any of our patients after a median follow-up duration of 17 months (range 15-41).Conclusions Wide local excision together with the skull invaded by tumors completely is effective surgical treatments for recurrent scalp DFSP; Use of scalp transferring technique could improve wound repair after complete tumor excision.

Thunder-fire wonder moxibustion is one of pressing moxibustion therapies and has a very good therapeutic effect on limb pains, furuncle-carbuncle and cold syndrome. To reveal the mechanism of clinical action of ancestors’ thunder-fire wonder moxibustion and seek the physical basis of its therapeutic advantage, this study, by a series of experiments, compared heat transfer regularities of thunder-fire wonder moxibustion versus pure moxa stick in simulated biological tissues under different conditions, preliminarily revealed heat radiation and heat transfer regularities of thunder-fire wonder moxibustion, tried to find pressing strength suitable for clinical operation of pressing moxibustion and had thoughts about changes in the clinical operation of past dynasties.

Objective:To investigate the expression of survivin and MMP-9 in the eutopic endometrium of patients with endometriosis(EMs).Methods:Eighty patients with endometriosis were selected as EMs group,four of them were in clinical stage Ⅰ,25 in stage Ⅱ,44 in stage Ⅲ and 7 in stage Ⅳ,47 in the proliferative phase and 33 in the secretory phase of the menstrual cycles.Twenty patients with hysteromyoma were selected as control group,including 7 patients in the proliferative phase and 13 in the secretory phase of the menstrual cycles.The expression levels of survivin and MMP-9 were determined by immuno-histochemical method.Image-Pro Plus 6.0(IPP),the special-purpose system for quantitative measurement of medical images,was used to analyse the results quantitatively.Results:Both survivin and MMP-9 were found positive in the plasma of the endometriosis.Both survivin and MMP-9 expression levels were higher in EMs group than those in the control group(P ＜0.01).Besides,survivin was over expressed with no relation to the menstrual cycle of the endometium.Survivin expression was higher in endometriosis stage Ⅱ than that of the stage Ⅲ(P ＞ 0.05).There was no difference in survivin expression within other stages(P ＞ 0.05).While MMP-9 expression had no difference between all the endometriosis stages(P ＞ 0.05).There was a close relationship between survivin and MMP-9 expression levels in 80 eutopic endometfium of endometriosis(r=0.262,t=2.860,P ＜ 0.05).Conclusion:The over expression of survivin and MMP-9 may play an important role in the pathogenesis of endometriosis.

Objective To explore the inhibitory effect and possible mechanisms of lianhuaqingwen capsules on radiation-induced acute lung injury in rats. Methods Rats were randomly divided into control group, radiation group and radiation plus lianhuaqingwen group, the control group and the radiation group rats were given 0. 9% sodium chloride solution, the radiation plus lianhuaqingwen group rats were given lianhuaqingwen 0. 9% chlorine sodium solution. HE staining was applied to test the lung tissue inflammation; quantitative RT-PCR and ELISA were used to measure the content of IL-6, TNF-α and MCP-1 in rats;immunohistochemical assay was taken to detect the infiltration of macrophage in lung tissues. Results The relative mRNA expression of IL-6, TNF-α and MCP-1 in the control, radiation model control and radiation plus Lianhuaqingwen groups were (0. 002 1±0. 000 20),(0. 006 6±0. 000 32),(0. 003 9±0. 000 22); (0. 003 7±0. 000 16),(0. 007 4±0. 000 33),(0. 005 5± 0.000 24);(0.001 4±0.000 15),(0.005 4±0.000 72),(0.003 2±0.000 17),respectively; the concentration (pg·mL-1) of IL-6,TNF-αand MCP-1 in the serum were (35. 2±10. 9),(111. 8±26. 1),(68. 2±15. 2); (229. 3±28. 5),(837. 5±57. 6), (566. 9±39. 8);(96. 85±8. 20),(314. 53±12. 76),(191. 32±10. 97),respectively; and the macrophages at high magnification field in each group were (59. 5±4. 3),(503. 9±25. 8)and (106. 2±12. 6), respectively. Lianhuaqingwen capsules significantly alleviated the lung inflammation in rats with radiation-induced acute lung injury,inhibited the accumulation of macrophage in lung tissue,reduced the expression of IL-6 and TNF-α,and decreased the content of MCP-1 in lung tissues and sera(P<0. 05). Conclusion Lianhuaqingwen capsules attenuated the lung inflammation developed in rats with radiation-induced acute lung injury through inhibiting the expression of MCP-1 and reducing the accumulation of macrophage in lung tissues.

Objective To investigate the radioprotective function of lianhuaqingwen (LHQW) in rat acute radiation-induced lung injury.Methods Totally 36 female Wistar rats were randomized into 3 groups as administered group (treated by LHQW plus radiation),radiation group irradiated with a single of 20 Gy in 6 MV X-ray by Elekta Synergy VMAT,and blank control group without radiation.Performance status (PS) was estimated during 31 d of LHQW instragastric administration.After rats being sacrificed at 1,14,28 d of LHQW adminstration,the pathomorphological changes were observed in trauma lung tissue,the cell number in BALF (Bronchoalveolar lavage fluid) was counted,the levels of TNF-α and IL-6 in serum were measured by ELISA,and TNF-α and IL-6 mRNA expressions in lung tissue were assayed by RT-PCR.Results After LHQW treatment,the PS of rat was significantly elevated with less inflammation in morphous,and the cell number in BALF was markedly decreased in compare with radiation alone group.Furthermore,the serum levels of TNF-α and IL-6 were obviously reduced (tTNF-α =7.372,2.891,tIL-6 =6.335,3.257,P ＜ 0.05) and the TNF-α and IL-6 mRNA levels in lung tissue were also decreased (tTNF-αmRNA =3.714,2.144,tIL-6mRNA =3.589,2.883,P＜0.05).Conclusions LHQW plays a protective role against acute radiation-induced lung injury in rats and the down-expressions of TNF-α and IL-6 may be involved.

Three kinds of PEGylated gold nanoparticles (PEG-Au NPs) with different surface charges are prepared by assembly of thiolated polyethylene glycol (HS-PEG) with different terminal groups including methoxy, amino or carboxyl on gold nanoparticle surface through sulfur-gold covalent bond, respectively.The experimental results of cell co-culturing and tail intravenous injection in mouse indicate that the biological behaviors of PEG-Au NPs are affected significantly by their surface charges.The cellular internalization amounts of PEG-Au NPs are following the order, positive charge ＞ neutral charge ＞ negative charge.The PEG-Au NPs are gradually transferred to liver and spleen from main organs through the circulation of blood after tail intravenous injection in mouse.The negatively charged PEG-Au NPs have the slowest hepatic clearance rate while the positively charged PEG-Au NPs can cause the strongest response of immune system in mice.

Insertional mutagenesis is a widely used method to determine the function(s) of a gene. To study the function(s) of the gene nsdAmgh in Streptomyces roseoflavus, a homologous recombination vector pSRNA2500 was structured in this paper. The recombination donor vector was then transformed into Strempomyces roseoflavus strain Men-myco-93-63 by conjugative transfer. The transformants were subjected to selection under the pressure of high temperature and appropriate antibiotics. As a result, several disrupted mutants of nsdAmgh gene, with a phenotype of Am(s)Km(r), were isolated and verified using PCR and Dot-blotting and Southern blotting hybridization methods. Functional analysis showed that the disrupted mutants of nsdAmgh had a two-fold higher inhibition against Verticillium dahlia Kleb than that of the wild strain Men-myco-93-63, which all will provide a new study route for future research about positive and negative regulator in Men-myco-93-63.
Genes, Bacterial ; Genetic Vectors ; Mutagenesis, Insertional ; Polymerase Chain Reaction ; Streptomyces ; genetics