Effects of parent-child painting and creative crafting therapy on preschool children with autism spectrum disorder and their mothers: a prospective randomized controlled trial / 中国当代儿科杂志

OBJECTIVES@#To study the effects of parent-child painting and creative crafting therapy on the core symptoms of preschool children with mild-to-moderate autism spectrum disorder (ASD) and the parenting stress and hope level of their mothers.@*METHODS@#A total of 56 preschool children with mild-to-moderate ASD and their mothers were divided into an experimental group and a control group using the block randomization method, with 28 pairs in each group. The subjects in the control group received an applied behavior analytic intervention and those in the experimental group received parent-child painting and creative crafting therapy in addition to the intervention in the control group. The intervention time was 20 weeks for both groups. Autism Behavior Checklist (ABC), Social Responsiveness Scale (SRS), Parenting Stress Index-Short Form (PSI-SF), and Herth Hope Index (HHI) were used to evaluate the core symptoms of children and the parenting stress and hope level of their mothers before and after 20 weeks of intervention.@*RESULTS@#Forty-nine child-mother pairs completed the study (25 pairs in the intervention group and 24 pairs in the control group). The children in the experimental group had significantly lower scores of social interaction, language, social communication, and social motivation and total scores of ABC and SRS compared with those in the control group (@*CONCLUSIONS@#The combination of applied behavior analytic intervention with parent-child painting and creative crafting therapy can more effectively improve the core symptoms and social interaction of preschool children with mild-to-moderate ASD, reduce the parenting stress of mothers and improve their hope level.

Curative effect of progressively integrated sandplay therapy on core symptoms and sleep management in preschool children with mild-to-moderate autism spectrum disorder / 中国当代儿科杂志

OBJECTIVE@#To explore the curative effect of progressively integrated sandplay therapy on core symptoms and sleep management in preschool children with mild-to-moderate autism spectrum disorder (ASD).@*METHODS@#A total of 50 mild-to-moderate ASD preschool children were randomly divided into an experimental group (n=25) and a control group (n=25). The control group was treated with structured teaching and auditory integration training (AIT). Based on these, the experimental group was treated with progressively integrated sandplay therapy. The Autism Behavior Checklist (ABC), Children Autism Rating Scale (CARS), Social Responsiveness Scale (SRS), Children's Sleep Habit Questionnaire (CSHQ) and emotion recognition tools were used to evaluate the overall curative effect before treatment and 3 months after treatment.@*RESULTS@#The experimental group had significantly lower scores of feeling, social interaction, somatic motor, language factor, total ABC and total CARS than the control group (P<0.05). The experimental group had also significantly lower scores of social cognition, social communication, social motivation, autistic behavior factor and total SRS than the control group (P<0.05). The experimental group had a significantly higher accurate rate of recognition of the upper facial expression and lower facial expression than the control group (P<0.05). The experimental group had significantly lower scores of bedtime resistance, sleep onset, sleep duration, night waking, daytime sleepiness, and total CSHQ than the control group (P<0.05). The experimental group had significantly reduced themes of wounding and significantly increased themes of healing after sandplay therapy (P<0.05).@*CONCLUSIONS@#Progressively integrated sandplay therapy can effectively improve the core symptoms and sleep quality of preschool children with mild-to-moderate ASD, which can be used as an early rehabilitation measure.

Expression of the MYL2 gene in the development of rat testis tissue / 中华男科学杂志

<p><b>Objective</b>To study the expression of the gene of myosin regulatory light chain-2 (MYL2) in the development of rat testis tissue.</p><p><b>METHODS</b>Using real-time PCR and immunohistochemistry, we determined the mRNA transcription level and protein expression of MYL2 in the rat testis.</p><p><b>RESULTS</b>The mRNA expression of the MYL2 gene changed in an age-dependent manner, reaching the highest value on postnatal day (PND) 2, then dropped rapidly till PND 8, increased slowly on PNDs 10 and 12, decreased on PND 14, rose slightly from PND 15 and rapidly on PNDs 20 and 25, and declined slowly from PND 65. Immunohistochemistry showed that the MYL2 protein was mainly expressed in testicular sperm cells.</p><p><b>CONCLUSIONS</b>The MYL2 gene may be involved in the proliferation of spermatogonial stem cells and the process of sperm cells developing into mature sperm.</p>

Effects of circulating microvesicles containing microRNA-27 a on blood brain barrier tight junction injury of ischemia stroke mice / 中国药理学通报

Aim To study the effect of circulating mi-crovesicles containing miR-27 a on blood brain barrier tight junction injury of ischemia stroke mice and its mechanism. Methods The middle cerebral artery occlusion mouse model was established, and the mi-crovesicles in the supernatant of 2 h of ischemic stroke brain tissues were separated by centrifugal ultrafiltra-tion method. The transmission electron microscopy was used to observe microvesicle morphology, and the di-ameter of microvesicles was detected. Based on the 2 h ischemia stroke mouse model, mice were injected via femoral vein with microvesicles at 5 mg · kg-1 . TTC staining was applied to detect infarction volume of is-chemia brain, while HE staining was applied to detect the expression change of tight junction protein occludin and claudin-5 . Western blot was subjected to detect occludin, claudin-5, TLR4, NF-κB and p38 protein expression. ELISA method was used to measure the contents of cytokines IL-1β and TNF-α. Results The microvesicle shape was approximately circular bi-lateral membrane structure, with an average diameter of 160 nm, which conformed to the morphological char-acteristics of microvesicles. Compared with the ische-mic stroke group, injection of microvesicles could ag-gravate the damage of brain tissues in ischemic mice, further increase the infarct volume and reduce the posi-tive staining areas of occludin and claudin-5 in ische-mia brain tissues. Meanwhile, the protein expressions of occludin and claudin-5 further decreased, and fur-ther up-regulated TLR4 and phosphorylation of NF-κB and p38 compared with the ischemia stroke group. Al-so, more content of IL-1βand TNF-αwere detected in ischemia stroke group injected with microvesicles com-pared with those in ischemia stroke group with signifi-cant difference, while the injection of antagomir-27a could alleviate brain damage and reduce the activation of TLR4, NF-κB and p38 in ischemia stroke mice. Conclusions Microvesicles containing miR-27 a could significantly attenuate brain injury in ischemia stroke mice, while aggravate the tight junction damage of ischemia brain. The mechanism might be correlated with the up-regulation of the expression of TLR4 , the phosphorylation of NF-κB, p38, and the release of cy-tokines IL-1β and TNF-α.

Clinical efficacy of bilateral MIS-LLIF via Quadrant channels for single-level degenerative lumbar spondylolisthesis / 局解手术学杂志

Objective To investigate the clinical efficacy and related complications of bilateral minimally invasive transforminal lumbar interbody fusion via Quadrant channels for the treatment of single-level degenerative lumbar spondylolisthesis.Methods Sixty selective pa-tients with single-level mild and moderate degenerative lumbar spondylolisthesis who underwent bilateral MIS-TLIF via Quadrant channels from August 2015 to May 2017 were retrospectived investigated.Operative time,blood loss,X-Ray exposure time,postoperative drainage and related complications were recorded.Lumbar functional improvement was defined as an improvement in the JOA.The VAS was also employed at pre and postoperation(6 months),to evaluate low back and leg pain.Intervertebral fusion was assessed by Bridwell evaluation criteria and the clinical outcome was assessed by the MacNab scale at 6 months.In addition,related complications also were be analyzed.Results All patients underwent a successful MIS-TLIF surgery,of whom 58 patients were followed up for 6 months.The operation time was 170~290 minutes (mean:220.17 minutes),intraoperative blood loss was 100~260 mL(mean:168.62 mL),intraoperative radiation exposure was 15~36 times (mean:25.60 times) and postoperative drainage was 50~163 mL(mean:103.43 mL).Different significance between 6 months post-operative follow-up and pre-operation was exhibited (P<0.05) in VAS scores (t=34.97,P=0.000).At 6 months post-operative follow-up,there were significant differences(P<0.05) compared with pre-operation in JOA scores(t=36.91,P=0.000).At 6 months post-operative follow-up,21 cases of grade 1 and 37 cases of grade 2 fusion were present as determined by the Bridwell evaluation criteria(100% fusion).MacNab scale as-sessment classified 2 patients having excellent clinical outcome and 54 patients in good clinical outcome(96.55% of patients had good to excel-lent results).Amomg 60 cases,complications observed in 6 patients,direct injuries to nerve root in 1 case,dural tears occurred in 1 case,super-ficial wound infection in 1 case,pedicle screw was pulled out during the reduction of slip because of severe osteoporosis in 1 case and leg numb-ness and pain in 2 cases. Conclusion Bilateral MIS-TLIF via Quadrant channels with less complications is a safe and effective approach for the treatment of single segment degenerative lumbar spondylolisthesis,but potential complications should be prevented as well.

Application of 3D printing in surgical approach choice for treatment of cervical vertebra dumbbell tumor / 局解手术学杂志

Objective To explore the application of 3D printing technology in surgical approach choice for cervical vertebra dumbbell tumor.Methods Retrospectively analyzed the clinical data of 17 patients with cervical vertebra dumbbell tumor who were admitted into our hospital from August 2012 and February 2017 and recieved 3D model printing.Three director surgeons chosen surgical approach for each pa-tient with or without 3D printing models.The surgical approach,combined with 3D printing and chosen by most surgeons,was selected as final surgical approach for patients.All the patients were evaluated by JOA score and Frankel grade before and after operation.Results The tumor of patients who selected surgical approach with 3D printing was completely resected.Without 3D printing models in preoperation,3 surgeons selected the same surgical approach for 8 patients,2 surgeons selected the same surgical approach for 9 patients.Compared with those who se-lected surgical approach with 3D printing models in preoperation,the 3 surgeons changed surgical approach in a total of 13 times.The surgical approach of 5 patients were changed,including 2 cases changed from posterior approach to lateral approach,2 cases changed from lateral ap-proach to posterior approach,and 1 case changed from lateral approach to anterior approach.Eventually,11 cases underwent posterior ap-proach,1 case underwent anterior approach,3 cases underwent lateral approach and 2 cases underwent far lateral approach.Conclusion Ap-plied of 3D printing in treatment of cervical vertebra dumbbell tumor can rebuild and print local anatomy by digitizing method.It can excel-lently show the tumor form,relationship of tumor and vertebrae,tumor and foramen intervertebrale,tumor and vessel.3D printing technology could help surgeon precisely and directly understand the operation area and select advanced surgical approach to reduce surgical risk and in -crease the resection rate of tumor.And it has an excellent display effect especially for complex tumors and abnormal blood vessels.

AppIication of poIymer interbody fusion cages / 中国组织工程研究

BACKGROUND: Interbody fusion is an effective approach in the spinal surgery which is traditionally performed via auto/allogenic bone grafting,and it has a number of shortcomings as well as specific therapeutic effects. In the past few years, polymer interbody fusion cages have been gradually applied in clinical practice, which provides more options for spinal surgeons. OBJECTIVE: To introduce the advantages and disadvantages of degradable/non-degradable polymer materials respectively by analyzing their materialogy characteristics, and to further predict their application prospects. METHODS: PubMed, Ovid and CNKI were searched by computer to retrieve articles concerning the materialogy characteristics and application situation of common biomedical polymer materials in the spinal surgery. The key words were "spinal surgery, fusion cage, engineering plastic, degradable high polymer material" in Chinese and English, respectively. RESULTS AND CONCLUSION: Up to now, a variety of polymer materials, especially degradable polymer materials, are still at the stage of exploring and primary research, and their in-depth studies, including animal and biomechanics experiments, are still expected to be done gradually. Except for PEEK interbody fusion cages, some polymer materials are still lack of large sample and multi-center clinical trials before further application.

Effect of guizhi plus gegen decoction on ultrastructural changes of intervertebral disc annulus fibrosus cells / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the effect of Guizhi plus Gegen Decoction (GGD) on ultrastructural changes of intervertebral disc annulus fibrosus cells.</p><p><b>METHODS</b>Rats' intervertebral disc annulus fibrosus cells were isolated and cultured using adherence wall screening method. After annulus fibrosus cells were intervened by GGD, the microstructure and ultrastructural features of untreated annulus fibrosus cells and annulus fibrosus cells treated by GGD containing serum at different concentrations were observed under optical microscope and electron microscope.</p><p><b>RESULTS</b>Under optical microscope, most annulus fibrosus cells showed irregular polygons and few in star shape with rich superficial ecphyma. The nuclei were oval, large and complete. Under electron microscope, most cells in the blank group were oval after intervened by GGD containing serum at different concentrations. The nucleus was large, deviated, and irregular, the heterochromatin scattered diffusely, partial mitochondria vacuolized, and rough endoplasmic reticulum dilated. In the low dose GGD group, increased mitochondria and condensed density could be seen. The rough endoplasmic reticulum were expanded, lipid drops or glycogen could be occasionally seen. In the middle dose GGD group, increased endoplasmic reticulum expansion and condensed density could be seen. More medium density protein sediment could be seen. Increased mitochondria with condensed density could be seen, showing irregular cystic form with various sizes nucleus. In the high dose GGD group, increased rough endoplasmic reticulum with obvious expansion could be seen. More high density protein sediment could be seen. The nuclei were deviated. More mitochondria could be seen with secretory granules in them.</p><p><b>CONCLUSIONS</b>After intervened by GGD containing serum at different concentrations, the ultrastructure of annulus fibrosus cells were manifested as follows: (1) The endoplasmic reticulum increased more in the middle and high dose GGD groups than in the blank group and the low dose GGD group. Greater density protein sediment occurred, especially in the high dose GGD group. (2) GGD played an important role in preventing ultrastructural changes induced by the degeneration of annulus fibrosus cells.</p>

Characteristics and risk factors of lymph node metastases in esophageal carcinoma / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To investigate the status of lymph node metastases (LNM) of esophageal carcinoma and to identify the risk factors.</p><p><b>METHODS</b>Clinical data of 308 patients who underwent esophagectomy with three-field lymphadenectomy during January 2006 and December 2010 were reviewed. Characteristics of LNM were studied.</p><p><b>RESULTS</b>The average number of dissected lymph nodes was 35.6 ± 14.5 in 308 patients. There were 197 patients(64%) had LNM. Logistic regression analysis showed that lymphatic vessel invasion(P=0.019) and deep tumor invasion(P<0.001) were risk factors of LNM. The highest LNM site was paratracheal node(25.0%). The incidence of cervical LNM was 14.1% in the middle thoracic carcinoma, higher than that of upper thoracic (7.3%) and lower thoracic (8.3%). Rate of LNM was lower in upper thoracic carcinomas than that in middle or lower ones(P=0.001). No significant difference of LNM was found among upper, middle and lower thoracic carcinoma for cervical or thoracic nodes. Lymphatic vessel invasion(P<0.001) and metastases in paratracheal lymph nodes (P=0.014) were risk factors for cervical LNM.</p><p><b>CONCLUSIONS</b>LNM of esophageal carcinoma can be found in both directions vertically and skipped metastasis. Paratracheal lymph nodes involvement is an indicator for cervical lymphadenectomy in thoracic esophageal carcinoma.</p>

Extraction, isolation and purification for ginkgolide B / 中国中药杂志

<p><b>OBJECTIVE</b>To establish a simple extraction, isolation and purification method for ginkgolide B from ginkgo leaf.</p><p><b>METHOD</b>The optimum conditions of extraction, isolation and purification were studied by taking the transfer rate of ginkgolide B as index.</p><p><b>RESULT</b>Ginkgo leaf was extracted with 70% ethanol for three times, the extracts were concentrated to remove ethanol and diluted by water till the crude drug density reached 0.1 g x mL(-1). The dilution was adsorbed with HPD-450 macroporous resin. The impurities were eluted with 20% ethanol and ginkgolide B was eluted with 80% ethanol. Then the 80% ethanol eluant was concentrated and crystallized. Finally the crude crystals were recrystallized with isopropanol. The purity of the ginkgolide B recrystallization was 95%.</p><p><b>CONCLUSION</b>The process was stable and easy to operate, which was suited to industrialized production.</p>

Comparison of mini-probe endoscopic ultrasonography with computed tomography in preoperative staging of esophageal cancer / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To compare mini-probe endoscopic ultrasonography (MCUS) with computed tomography (CT) in preoperative T and N staging of esophageal cancer, and to find out the MCUS parameters to judge lymph node metastasis for esophageal cancer.</p><p><b>METHODS</b>Thirty-five patients received both MCUS and CT preoperatively, on both of which the T and N stages were determined. The accuracy, sensitivity, specificity, positive predicting value and negative predicting value were compared with the postoperative pathological results.</p><p><b>RESULTS</b>The accuracy of MCUS was 85.7% in T staging and 85.7% and 80.0% in N staging by the two different methods, which were 45.7% and 74.3%, respectively, by CT.</p><p><b>CONCLUSION</b>MCUS is better than CT in preoperative staging for esophageal cancer. The ratio of short to long axis (S/L) combined with short axis is a useful way to determine lymph node metastasis.</p>

Clinical study on the treatment of male immune infertility by Huzhangdanshenyin / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the efficacy of the Chinese medicine Huzhangdanshenyin on male immune infertility, and provide an effective therapy for this disease.</p><p><b>METHODS</b>Ninety men with immune infertility were selected as the research subjects and randomly divided into two groups, 60 in the treatment group, treated by Huzhangdanshenyin, and the other 30 in the control, treated by prednisone, both for 3 months. The improvement of clinical symptoms, immunologic indexes (antisperm antibodies in serum and seminal plasma) and sperm indexes (semen liquefied duration, motility, viability, density and abnormal morphology rate) were observed and the results analyzed.</p><p><b>RESULTS</b>The total antisperm antibody reversing ratio of the treatment group was higher than that of the control (P < 0.01), especially the serum antisperm antibody reversing ratio. There were significant differences in the clinical cure rate and total validity rate between the treatment group and the control (P < 0.01). After the treatment, the markers of the clinical symptoms (P < 0.01), especially the serum antisperm antibody reversing ratio. There were significant differences in the clinical cure rate and total validity rate between the treatment group and the control (P < 0.01). After the treatment, the markers of the clinical symptoms were lower (P < 0.01), and the improvement of the clinical symptoms was better in the treatment group than in the control (P < 0.01), especially the symptoms of pain in the back and knees, distending and bearing-down sensation of the perineum and testis, hypersexuality and topalgias. Compared with pre-treatment, sperm motility and viability of the treatment group significantly improved (P < 0.01), and so did sperm density (P < 0.05). However, there were no significant differences in sperm density, semen liquefied duration, abnormal morphology rate and pH (P > 0.05) before and after the treatment.</p><p><b>CONCLUSION</b>The Chinese medicine Huzhangdanshenyin works more effectively than prednisone in the treatment of male immune infertility. It could improve the antisperm antibody reversing ratio, clinical symptoms and signs and ameliorate sperm indexes with no obvious advierse effects.</p>

Effects of carbaryl production exposure on the sperm and semen quality of occupational male workers / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To analyse the male reproductive toxicity of carbaryl.</p><p><b>METHODS</b>Thirty-one male carbaryl exposure workers and 46 male administrators in the office in a pesticide factory were selected as the exposure group and internal control group respectively, and 22 male administrators in a center for disease control and prevention were served as the external control group. In order to evaluate the exposure levels, the concentrations of carbaryl, methyl isocyanate, ammonia and phenol in the ambient air of the work place in these three groups were monitored simultaneously for three consecutive days. Moreover, three workers in the exposure group and the external control group were selected to evaluate the amount of carbaryl of individual and dermal contamination for three consecutive days. After the semen were collected according to the standard method, the workers'semen qualities were analysed with WHO method, the sperm morphology and the sperm motility were evaluated using micro-cell slide spectrum technology and computer assisted sperm analysis (CASA) respectively.</p><p><b>RESULTS</b>In the exposure group, the concentrations of carbaryl and phenol (52.41 mg/m(3) and 0.08 mg/m(3) respectively) were significantly higher than those in the internal and external control group (P < 0.01 or P < 0.05). Furthermore, in the carbaryl exposure area the geometric mean concentration of carbaryl with the individual sampling was 7.38 mg/m(3), and the geometric mean of dermal contamination detected in the carbaryl exposure area was 862.47 mg/m(2). Carbaryl was not found in the external control area (P < 0.01). The seminal volume [(2.39 +/- 1.44) ml] and the sperm motility [(1.77 +/- 0.61) grade] were significantly lower than those in the external control group (P < 0.05), and sperm motion parameters such as linearity (LIN, 39.89% +/- 6.00%), straightness (STR, 71.51% +/- 11.22%), straight line velocity [VSL, (26.29 +/- 7.84) microm/s] and beat cross frequency [BCF, (3.99 +/- 1.55) Hz] were lower than those in the internal and external control group (P < 0.05), while the abnormal rates of viscidity, sperm motility and total aberration rate were higher than those in the external control group (P < 0.05).</p><p><b>CONCLUSION</b>Occupational exposure to carbaryl production can affect the workers'sperm and semen quality to certain extent.</p>

Pharmacokinetics of oxymatrine and its metabolite in beagle dogs by LC-MS / 中国中药杂志

<p><b>OBJECTIVE</b>To establish LC-MS method in the determination of oxymatrine and its metabolite in plasma and investigate their pharmacokinetics in beagle dogs.</p><p><b>METHOD</b>Lichrospher C18 column (4.6 mm x 250 mm, 5 microm) was used as the analytical column maintained at 25 degrees C. The mobile phase consisted of 10 mmol x L(-1) CH3COONH4 and CH3OH (25:75). Flow rate was 1 mL x min(-1). Electrospray ionization (ESI) was carried out. The ESI ion source was set in positive ion polasity mode. The selective ion monitoring (SIM) was set at m/z 265.1 and 249.2.</p><p><b>RESULT</b>The linearity ranged from 2 to 5000 ng x mL(-1) (r = 0.9991). The detection of oxymatrine and its metabolite were 0.6 and 0.3 ng x mL(-1). The RSD(%) within day and between day was less than 4.7%. The recovery of this method was more than 96.5%. The disposition was conformed to a two-compartment model. The T(1/2), Tmax, Cmax, MRT, AUC(0-->24 h) of oxymatrine were (5.5+/-1.58) h, (1.0+/-0.30) h, (2418.3 +/-970.78) ng x mL(-1), (3.2+/-0.64) h, (5797.4+/-908.16) ng x mL(-1) x h accordingly. The corresponding T(1/2), Tmax, Cmax, MRT, AUC(0-->24 h) of matrine were (9.8+/-2.77) h, (1.9+/-1.09) h, (1532.4+/-494.86) ng x mL(-1), (4.4+/-1.97) h, (5530.5+/-1042.65) ng x mL(-1) x h.</p><p><b>CONCLUSION</b>This assay was highly sensitive, rapid, simple and specific enough for determining concentrations of oxymatrine and its metabolite matrine in plasma of beagle dog.</p>

Study on the extraction recovery of ginsenoside Re in plasma by different solid-phase cartridges / 中国中药杂志

<p><b>OBJECTIVE</b>To optimize the solid-phase extraction method by comparison of the extraction recovery of ginsenoside Re plasma samples.</p><p><b>METHOD</b>After extracted by different solid-phase cartridges with water, acetonitrile, and different content methanol elution, the plasma samples were analyzed on an Zorbax SB-C18 column with acetonitrile-water gradient elution. From the recovery achieved, the best solid phase cartridge was found.</p><p><b>RESULT</b>This method consists of using 40% methanol as the wash solvent, and 80% methanol for the elution. Among the three kinds of solid-phase being tested, Waters Oasis HLB cartridge was found to be the best one.</p><p><b>CONCLUSION</b>The average extraction recovery of the Waters Oasis HLB cartridges was between 103%-113%, it can be used in the analysis of ginsenoside Re in plasma samples.</p>

Fermentation, purification and identification of recombinant RGD-Hirudin / 生物工程学报

Recombinant RGD-Hirudin ( r-RGD-Hirudin ) has double functions: anti-thrombin activity and anti-platelet aggregation activity. To identify these functions, the expression plasmid, RGD-Hirudin-pPIC9K, was constructed by inserting cDNA of RGD-hirudin in yeast expression vector pPIC9K. The high expression clone was gained after screening. This clone was fermented for 3 days. The r-RGD-hirudin was secreted into the culture. It was ultra-filtrated from culture supernatant, then after gel filtration chromatography and anion exchange chromatography, the purified r-RGD-hirudin was gained. Its purity was larger than 97% and its specific activity was 12 000 ATU/mg. The yield per liter culture of purified r-RGD-hirudin was 1 g and overall recovery yield was more than 75% . The purified r-RGD-hirudin was identified by reductive SDS-PAGE, anti-thrombin activity assay, anti-platelet aggregation assay, LC/MS and isoelectrofocusing assay. It is proved that r-RGD-Hirudin is ramification of wt-Hirudin and it has anti-thrombin activity and anti-platelet aggregation activity.

Study on technology of extracting geniposidic acid from Eucommia ulmiodes and its application / 中国中药杂志

<p><b>OBJECTIVE</b>To study the optimum technology of extracting bioactive component named geniposidic acid in Eucommia Ulmiodes by orthogonal test.</p><p><b>METHOD</b>An orthogonal test table L16(4(5)) was used with the factors of extracting time, temperature, times, ethanol consistency and the ratio of solid to liquid. The content of geniposideic acid was determined by RP-HPLC.</p><p><b>RESULT</b>The optimum technology was: temperature 50 degrees C, time 1 h, once, consistency 80%, and the ratio of solid and liquid 1:12. Under the condition, the best material of Eucommia Ulmiodes for extracting geniposidic acid was chosen.</p><p><b>CONCLUSION</b>The technology can provide some theoretic advice for industry.</p>

Preparation and functional study of an adenovirus vector expressing human plasminogen kringle 5 gene / 生物工程学报

Tumor angiogenesis plays a pivotal role in the progress of tumor. Among the various endogenous angiogenic inhibitors discovered, the human plasminogen kringle 5 (K5) has been demonstrated to be a potential inhibitor of the proliferation and migration of vascular endothelial cells in vitro. The replication-incompetent adenovirus (Ad) vector Adeno-X-CMV-K5 (Ad-K5) (where CMV is cytomegalovirus) was constructed and its antiangiogenic effect was tested on vascular endothelial cell and tumor cell. For the construction, the K5 cDNA was fused in-frame with human plasminogen signal sequence and inserted into the eukaryotic expression vector pcDNA3 to form pcDNA3K5. The recombinant plasmid was subcloned into the shuttle plasmid pShuttle under the control of the constitutive CMV immediate-early promoter. The plasmid carrying the cDNA for K5 (pShuttleKS) was then recombined with the Adeno-X viral DNA and transformed into E. coli DH5alpha. The resultant recombinant plasmid pAd-K5 was transfected into human embryonic kidney (HEK) 293 cells with liposome. The adenovirus expressing human plasminogen kringle 5 (Ad-K5) was successfully packaged and propagated in 293 cells, as detected by the cytopathic effect (CPE) on the cells, and the viral titer in the supernatant was 5 x 10(8) pfu/mL by plaque assay. Both human umbilical vein endothelial cell line ECV304 and human breast carcinoma cell line MDA-MB-231 were infected with Ad-K5 and Ad-LacZ, which was used the negative control, and assayed by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. Compared with uninfected control and Ad-LacZ infected control, Ad-K5 infected group at 80 MOI (multiplicity of infection) significantly inhibited ECV304 proliferation; the difference between uninfected control and Ad-LacZ infected control was not significant. In contrast, there was no significant difference in the proliferation of MDA-MB-231 among all the treatments. In addition, the Ad-K5 at 100 MOI inhibited the differentiation and tube formation of ECV304 on ECMatrix gel. These results suggested that the recombinant replication-defective Adenovirus expressing human plasminogen kringle 5 inhibited the proliferation, differentiation and tube formation of ECV304 and had no effect on the proliferation of MDA-MB-231. Adenovirus mediated human plasminogen kringle 5 gene therapy may be a potential treatment of cancer through angiogenesis inhibition.

Analysis of impurity in caderofloxacin / 药学学报

<p><b>AIM</b>To analyse the main impurity of caderofloxacin.</p><p><b>METHODS</b>The impurity of caderofloxacin was analysed and determinated by RP-HPLC/ESI/MS with a Zorbax SB-C18 (150 mm x 4.6 mm ID, 5 microns) column. The mobile phase was acetonitrile-0.5% acetic acid solution (17:83). A compound was synthesized: 1-cyclopropyl-8-(difluoromethoxy)-6-fluoro-1, 4-dihydro-7-(1-piperazinyl)-4-oxo-3-quinoline carboxylic acid (DMCA). Its HPLC chromatogram, UV and MS spectrum were compared with those of the impurity in caderofloxacin.</p><p><b>RESULTS</b>The molecular weight of the impurity was 14 less than that of caderofloxacin. It means the impurity was a CH2-group less than caderoflixacin. The tR, UV and MS of DMCA were the same as those of the impurity in caderofloxacin.</p><p><b>CONCLUSION</b>Based on the tR (HPLC), UV and MS, the impurity of caderofloxacin is confirmed as DMCA.</p>

Determination of first-order structure of somatostatin by electrospray ionization mass spectrometry / 药学学报

<p><b>AIM</b>To determine the molecular weight and first-order structure of somatostatin.</p><p><b>METHODS</b>The molecular weight of somatostatin was determined by electrospray ionization mass spectrometry. Somatostatin was deoxidized by 2-mercaptoethanol. A series of typical fragment ions of deoxidized product were obtained by insource collision-induced dissociation (CID).</p><p><b>RESULTS</b>The m/z of quasi-molecular ion [M + H]+ of somatostatin was 1,637.8 and [M + Na]+ was 1,659.5. The m/z of double-charge ion [M + 2H]2+ was 819.5 and [M + H + Na]2+ was 830.3. It showed that the molecular weight of somatostatin was 1,636.7. The y and b series of fragment ions of deoxidized product were obtained by adjusting the fragmentor voltage. It was determined that the first-order structure of deoxidized product of somatostatin was A-G-C-K-N-F-F-W-K-T-F-T-S-C.</p><p><b>CONCLUSION</b>The molecular weight and first-order structure of somatostatin were confirmed.</p>