OBJECTIVE:To explore a new extraction method of baicalin. METHODS: A new continuous circular extraction equipment was adopted to extract baicalin. L9(43) orthogonal experiment was conducted to optimize the extraction process taking the content of total flavonoids as index with the number of period of batching cycle of solvent, extraction temperature and distillation time as factors. The new continuous circular extraction was compared with the aqueous extraction and acidic deposition method in terms of the yield of extractum, content of total flavonoids and baicalin content and the color of the extract. RESULTS: The best technical conditions for the new extraction method of baicalin were as follows: 2.5 periods in batching cycle of solvent, 50 ℃ in extraction temperature, and 6 h in distillation time. The new continuous circular extraction was significantly better than the aqueous extraction in terms of contents of total flavonoids and baicalin and the color of the extract. CONCLUSION: The new continuous circular extraction method is applicable for the extraction of baicalin.

Objective By detecting the variation of long-chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD) DNA methylation in preeclampsia-like mouse models generated by different ways, to explore the roles of multifactor and multiple pathways in preeclampsia pathogenesis on molecular basis. Methods Established preeclampsia-like mouse models in different ways and divided into groups as follows: (1) Nw-nitro-L-arginine-methyl ester (L-NAME) group: wild-type pregnant mouse received subcutaneous injection of L-NAME;(2) lipopolysaccharide (LPS) group:wild-type pregnant mouse received intraperitoneal injection of LPS; (3) apolipoprotein C-Ⅲ (ApoC3) group: ApoC3 transgenic pregnant mouse with dysregulated lipid metabolism received subcutaneous injection of L-NAME;(4)β2 glycoprotein I (β-2GPI) group:wild-type pregnant mouse received subcutaneous injection ofβ-2GPI. According to the first injection time (on day 3, 11, 16 respectively), the L-NAME, LPS and ApoC3 groups were further subdivided into:pre-implantation (PI) experimental stage, early gestation (EG) experimental stage, and late gestation (LG) experimental stage.β-2GPI group was only injected before implantation. LCHAD gene methylation levels in placental were detected in different experimental stage. Normal saline control groups were set within wild-type and ApoC3 transgenic pregnant mice simultaneously. Results (1) CG sites in LCHAD DNA:45 CG sites were detected in the range of 728 bp before LCHAD gene transcription start site, the 5, 12, 13, 14, 15, 16, 19, 24, 25, 27, 28, 29, 30, 31, 32, 34, 35, 43 CG sites were complex sites which contained two or more CG sequences, others were single site which contained one CG sequence. The 3, 5, 6, 11, 13, 14, 18, 28 sites in L-NAME, LPS, ApoC3 and β-2GPI groups showed different high levels of methylation; the 16, 25, 31, 42, 44 sites showed different low levels of methylation; other 32 sites were unmethylated. (2) Comparison of LCHAD gene methylation between different groups:the methylation levels of LCAHD gene at 3, 11, 13, 14, 18 sites in L-NAME, LPS, ApoC3 andβ-2GPI groups were significantly higher than those in the normal saline control group (P<0.05); and the methylation levels of 42, 44 sites in these groups were significantly lower than those in the normal saline control group (P<0.05). (3) Methylation of LCHAD gene at the same site between different experimental stages: ① The 3, 11, 18 sites of EG experimental stage was significantly lower than PI and LG experimental stage in L-NAME group (P<0.05);the 3, 11, 18 sites of PI experimental stage was significantly lower than EG and LG experimental stage in LPS group (P<0.05);these sites of PI experimental stage was significantly higher than EG and LG experimental stages in ApoC3 group (P<0.05).②The methylation of site 5 in L-NAME and LPS groups were significantly higher than that of the normal saline control group (P<0.05), and the LG experimental stages were significantly higher than other stages, but in ApoC3 group , only PI and EG stages were significantly higher than the normal saline control group (P<0.05).③At site 6 in L-NAME group which showed high methylation level was significantly higher than the same site in other groups which showed low methylation level (P<0.05).④At 13, 14 sites, earlier preeclampsia onset caused a lower methylation level in L-NAME group, but PI experimental stage was significantly higher than EG and LG experimental stages in LPS group (P<0.05), EG experimental stage was significantly higher than PI and LG experimental stages in ApoC3 group (P<0.05). ⑤ At site 28, earlier preeclampsia onset caused a higher methylation level in L-NAME group, but PI experimental stage was significantly lower than EG and LG experimental stages in LPS group (P<0.05), EG experimental stage was significantly higher than PI and LG experimental stages in ApoC3 group (P<0.05).⑥The 16, 25, 31 sites in ApoC3 group were significantly higher than other groups (P<0.05). ⑦ At site 42 in β-2GPI group was unmethylated, but it in other groups showed low methylation level, the methylation level of site 42 inβ-2GPI group was significantly lower than that in other groups (P<0.05). Conclusions The methylation of 6 and 42 CG sites may be related to LCHAD gene expression in placenta of L-NAME and β-2GPI induced preeclampsia-like models respectively;LCHAD gene expression and DNA methylation may not have obviouscorrelation in LPS and ApoC3 induced preeclampsia-like models. Differences exist in LCHAD DNA methylation in preeclampsia-like models generated by different ways, revealed a molecular basis to expand our understanding of the multi-factorial pathogenesis of preeclampsia.

Simple sequence repeat (SSR) was used to investigate the genetic diversity and structure of Dendrobium officinale. A total of 15 primer pairs with stable and repeatable polymorphism were screened out from 60 SSR primer pairs developed by the method of microsatellite enrichment by magnetic beads. Forty-eight samples of Dendrobium officinale were analyzed in genetic polymorphism. These loci were polymorphic and displayed 3 to 9 alleles per locus with a mean number of 6.1. The observed and expected heterozygosities ranged from 0.60 to 0.85 and from 0.49 to 0.85 respectively. The polymorphic information content (PIC) of each SSR locus varied from 0.437 to 0.829 with an average of 0.702. Fifteen primer pairs were used in Dendrobium cross-species amplification and totally 13 primer pairs were proved to have the transferability in D. officinale related species. In addition, 500 tissue culture plantlets of D. officinale were tested for purity identification by means of PCR amplification with four SSR primer pairs. The results showed that SSR technique is a feasible, simple and inexpensive method for determining adulterants in germplasm identification.

Objective To investigate the prevention and management of anastomotic leakage in patients with middle and lower rectal cancer after neoadjuvant therapy.Methods In this study,50 cases of middle or lower rectal carcinoma underwent anus preserving radical resection and anastomosis after a 4 cycles of chemotherapy with FOLFOX and 46 Gy(2 Gy×23)radiotherapy.Anastomotic leakage,the prevention and management of this complication were analyzed retrospectively.Results All these 50 patients received low anterior resection after chemoradiotherapy,among the 19 cases receiving prophylactic ileac stoma simultaneously,there was no anastomotic leakage,whereas anastomotic leakage developed in 4 out of the 31 patients who did not receive prophylactic ileac stoma,including concomitant rectovaginal fistula in 2 cases.All these fistulae were cured conservatively.Conclusion Concomitant treatment of FOLFOX regimen and RT in neoadjuvant setting of rectal cancer could increase the rate of anal interior sphincter reservation,prophylactic ileac stoma helps to reduce the rate of anastomotic leakage after a anus-preserving low anterior resection of the cancer.

Objective To study the effects of compound preparations which contain docosahexenoic acid (DHA),soybean lecithin,and vitamin A on memory ability of children and to compare the difference between two compound preparations that contain DHA from marine algae and from fish oil.Methods Totally 160 11-12-year-old healthy children who were studying in a primary school in Baoshan District of Shanghai were enrolled in this study.All the subjects signed the informed consent form.Subjects were randomly divided into three groups with random numbers:marine algae DHA group(n=53),fish oil DHA group(n=53),and control group(n=54).Subjects in the marine algae DHA group were given compound preparation which contained DHA form marine algae,soybean lecithin,and vitamin A;subjects in fish oil DHA group were given compound preparation which contained DHA form fish oil,soybean lecithin,and vitamin A.The dose of DHA(200 mg DHA per capsule)and other components in the two groups was equal.Subjects in the control group were given a placebo with same appearance.The trial lasted 30 days.Each subject took a capsule per day.Immediately before and after the trial,subjects were tested by using the clinical memory scale compiled by the Institute of Clinical Psychology of the Chinese Academy of Sciences.Results Before the trial.there was no difference amongthree groups in terms of all items of clinical memory scale or memory quotient(all P>0.05).After the trial,except for associative learning(both P>0.05),the other items of the clinical memory scale and memory quotient in both marine algae DHA group and fish oil DHA group were significantly higher than those of control group(all P<0.05).No significant difference was noted between the marine algae DHA group and fish oil DHA group for all items of the clinical memory scale or memory quotient(all P>0.05).Conclusions DHA compounds can impreve the memory ability of children.DHAs with different sources have similar effect on memory ability.

Objective To investigate the effects of tuina manipulation on creep rate and maximal breaking stress of contractural Achilles tendons of rabbits. Methods Twenty-four rabbits were randomly divided into contrac-tural Achilles tendon model group, free activities group and tuina manipulation group. The opposite legs of model group were used as normal controls. Creep rate and maximal breaking stress of Achilles tendons were compared among the 4 groups. Results The average creep rate in tuina manipulation group approximated to that in normal group and was lower than that in model and free activities groups. The average maximal breaking stress in tuina manipulation group approximated to that in normal group and was higher than that in model and free activities groups. Conclusion Tuina manipulation can promote the creep rate and maximal breaking stress of contractura Achilles tendon approximately to normal level. Tuina manipulation may be useful in rehabilitation.

Objective To summarize the clinical experience of ESWL under B ultrasound guidance in treatment of urinary stones.Methods The clinical data of 20 625 patients who underwent B ultrasound-guided ESWL between August 1995 and July 2011 were retrospectively analyzed.Of the stones,8659cases were in the kidney,11 712 cases were in the ureter,and 254 cases were in the bladder.And 1965stones were radioparent.Results The stone fragmentation rates of the kidney,ureter and bladder stones were 96.27％ ( 8336/8659),97.56％ ( 1 1 426/1 1 712) and 99.21％ ( 252/254),respectively.The total stone fragmentation rate was 97.04％ (20 014/20 625 ).The 3-month stone-free rates of the kidney,ureter and bladder stones were 86.63％ (7501/8659),95.32％ ( 11 164/11 712) and 97.24％ (247/254),respectively.The total 3-month stone-free rate was 91.69％ (18 912/20 625).The stone fragmentation rate of radioparent stones was 97.91％ ( 1924/1965 ),and the 3-month stone-free rate was 94.40％ ( 1855/1965).Conclusion B ultrasound-guided ESWL is safe and effective in the treatment of urinary stones.

This study aimed to examine whether ophiopogonin D (OP-D) is capable of protecting cardiomyocytes against DOX-induced injury and the mechanisms involved. H9c2 cells were cultured. MTT assay was used to evaluate cell viability and toxicity. Mito-tracker as fluorescence probe was used to measure ROS content raised from mitochondria. The mRNA and protein expression of ATF6alpha, GRP78 and CHOP were analyzed using real-time PCR and Western blotting, respectively. The results showed that a significant endoplasmic reticulum stress (ERS) was induced upon exposure of H9c2 cells to DOX as indicated by the increase in the expression of ERS related proteins, which was paralleled with the accumulation of reactive oxygen species (ROS) and decrease in the viability of H9c2 cells. Whereas, DOX-induced ROS accumulation and up-regulation of ERS related proteins were partially abolished by pretreatment with OP-D. Consequently, a DOX-induced ERS was mitigated by application of OP-D. Similarly, DOX-induced decrease in cell viability was partially attenuated by either inhibiting CHOP or pretreatment with N-acetylcysteine (NAC), an antioxidant. Moreover, cardiac ultrastructural abnormalities seen in mouse receiving DOX injections were obviously ameliorated by pretreatment of OP-D. Taken together, the present study proved that OP-D protects cardiomyocytes against DOX-induced injury, at least in part, through reducing ROS accumulation and alleviating ERS.

Objective To fabricate a silk fibroin scaffold with biomimetic oriented microstructure using the directional crystallization technology, and to evaluate the possibility of application to the cartilage tissue engineering. Methods The silk fibroin scaffold with biomimetic oriented microstructure was made by the directional crystallization technology. The structure of scaffold was observed by the scanning electron microscope, and the pore size, porosity and mechanical properties were calculated. Adipose-derived stem cells were isolated from rabbit, and the passage 3 was seeded into the scaffold. The cell proliferation was detected by CCK8 method. The cell adhesion ability was observed by HE staining and scanning electron microscope. The cell viability was observed by LIVE/DEAD staining. Results The scanning electron microscopy showed that the parallel microtubule-like structure can be seen arranged in longitudinal section of the scaffold, which had a uniform directivity, and also the elliptical or circular pore structure in cross-section. The scaffold had a good pore interconnectivity with (112.43±12.65) μm pore diameter of the cross-section, (90.25±2.05)%porosity and (52.48±5.78) kPa the compression modulus. HE staining and scanning electron microscopy demonstrated that the cells uniformly adhered to the surface and inner pore, and which secreted lots extracellular matrix distributed in the oriented microstructure. Results of CCK8 showed that the good cell proliferation on the scaffold, and the LIVE/DEAD staining indicated that the cells were maintained good viability. Conclusion The silk fibroin scaffolds have a biomimetic oriented microstructure, and show good pore diameter, porosity, biocompatibility and biomechanical properties, which made it a suitable candidate as an alternative scaffold for cartilage tissue engineering.

Objective To investigate the effect of resveratrol on miRNA-106b in Alzheimer′s disease ( AD ) animal model.Methods Fifty Kunming male mice were divided into five groups by completely randomized block sampling.The five groups included three dosage resveratrol groups , an AD model group and a control group.The AD models were established in one month prior to treatments. Subsequently, from the 31st day various doses of resveratrol were provided intragastricly for 60 days.Then the memory function was observed by the step-down test.Meanwhile, the varying expressions of APP , P62, ApoA1, miRNA-106b, ABCA1 were tested in each group to determine whether there is the binding site for miRNA-106b in APP 3′UTR sequence.Results Compared with the control group by step-down test, the memory function of the AD model group mice decreased in different degree , which in the drug treatment group was higher than that in the model group (P<0.05).Compared with the AD group, the expression of APP (1.131 ±0.035) in the drug treatment group was higher than that in the model group (0.652 ± 0.026), while the P62 (0.412 ±0.022) and ApoA1 (0.534 ±0.032) were lower than the model group ( all P<0.05 ).High and medium dose groups of resveratrol treatment reduced varying degrees of APP (0.733 ±0.018,0.929 ±0.019,F=177.733) levels, and increased P62(0.954 ±0.035,0.633 ±0.015, F=434.5 ) and ApoA1 ( 1.042 ±0.051, 0.824 ±0.034, F=286.582 ) levels ( all P<0.05 ).The expression of miRNA-106b (0.464 ±0.313) and ABCA1(0.293 ±0.042) in the model group was lower than that in the control group (miRNA-106b 1.064 ±0.032, F=238.159; ABCA1 0.781 ±0.027,F=341.61;both P<0.05).The miRNA-106b (0.843 ±0.034, 0.601 ±0.012) and ABCA1 (0.882 ± 0.025, 0.624 ±0.036) levels in the high, medium dose resveratrol treatment groups increased to different extent ( both P<0.05 ).After the drug treatment , luciferase reporter vector experiments showed that the APP 3′UTR sequence contains the binding site of miRNA-106b.Conclusions APP is one of the target genes of miRNA-106b.Resveratrol is capable of improving AD by enhancing the expression of miRNA-106b and down-regulating the target genes including APP , P62 and ApoA1.This provides a new theoretical basis for the clinical treatment of AD.