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Objective:To investigate overall quality status of rose bengal agar culture medium for drug control in China to provide technical support for the drug standard revision. Methods:Four quality control microorganisms, namely Aspergillus niger, Candida al-bicans, Staphylococcus aureus and Escherichia coli, were used to evaluate the quality of rose bengal agar culture medium from 7 domes-tic and foreign manufacturers. Results:Including the control medium, the pH value of all the 8 media was different. The growth rate of Aspergillus niger and Candida albicans in the seven tested media were all above 0. 7 without significant difference (P>0. 05). The selectivity factors of Staphylococcus aureus in the seven media and the control medium was more than 6. The eight media showed differ-ences in the selectivity factors of Escherichia coli. Among them, A, B, C, E, G and the control medium had no inhibitory ( selectivi-ty) , while D and F showed certain selectivity. Conclusion:The rose bengal agar media from different manufacturers for drug control show similar growth-promoting ability with different pH value and selectivity to Escherichia coli.

Objective In order to study the value of drug treatment for leukemia, quercetin was delivered to the cultured NB4 cell line. MethodsUsing RT- PCR and western blot, we studied the expression of mutant p53 gene and protein after inducing 24, 48, 72, 96 and 120h by the quercetin with different concentrations (30, 60, 90μmol/L). Results We verified the results that quercetin with concentration of 30 ～90μmol/L could successfully inhibit the expression of mutant p53 protein, but the mutant p53 gene did not have. ConclusionThat data suggested that quercetin would be an effective method for the therapy of leukemia.

Background Diabetes is one of the risk factors that leads to corneal neuropathy.Silent signal regulatory factor 1 (Sirt1) plays an important role in glucose metabolism,lipid metabolism,regulation of insulin secretion and is closely related to the nervous system disease.The relationship between Sirt1 and diabetic corneal neuropathy is not fully understood.Objective This study was to detect the expression of Sirtl in cornea and trigeminal ganglion with type 1 diabetes model mice and explore the association of Sirt1 expression with diabetic corneal neuropathy.Methods Eight C57BL/6-Ins2Akita/J male mice and eight wild-type C57BL/6 male mice in the same litter were selected as type 1 diabetes model group and control group,respectively.The mice of two groups were sacrificed in overdose anesthesia method at 12-month old.Histological examination of cornea and trigeminal ganglion was performed using hematoxylin and eosin staining.Expression and localization of Sift1 protein in cornea and trigeminal ganglion were detected using immunohistochemistry.Western blot assay and fluorescine quantitative PCR were respectively used to quantitatively analyze the expression of Sirt1 protein and Sirt1 mRNA.Results Trigeminal ganglion cells were uneven in size and shape with the loosened cellular arrangement and disorder neurofibrosis alignment,and the corneal epithelial cells were less in the C57BL/6-Ins2Akita/J mice,but the trigeminal ganglion cells and corneal epithelial cells were normal in wild-type C57BL/6 mice.Immunochemisty exhibited that Sirtl protein was expressed mainly in corneal epithelium and the expression of Sirtl protein was stronger in the C57BL/6 mice than that in C57BL/6-Ins2Akita/J mice.Fluorescine quantitative PCR assay showed that the gray scale value of Sirt1 mRNA in cornea in C57BL/6-Ins2Akita/J mice was lower than that of the wild-type C57BL/6 mice(0.56±0.03 vs.0.98±0.13) with significant difference (t =5.853,P =0.010).Western blot showed that the expression of Sirt1 protein in cornea was lower in C57BL/6-Ins2Akita/J mice than that of the wild-type C57BL/6 mice(0.78±0.017 vs.1.300±0.012) with significant difference(t =33.140,P =0.001).However,no significant differences were seen in the gray scale value of Sirt1 mRNA(2.45±0.18 vs.2.51±0.22) (t=0.587,P=0.599) and protein level(1.100±0.015 vs.1.110±0.017) (t =0.430,P=0.709) in trigeminal ganglion tissues between C57BL/6-Ins2Akita/J mice and wide-type C57BL/6 mice.Conclusions The corneal nerve and structure is abnormal in 12-month-old C57BL/6-Ins2Akita/J mouse.Sirt1 is involved in the pathogenesis of diabetic keratoneuropathy,suggesting that it may be a potential target.

Objective:To provide the basic data for the further revision of mycoplasma test method described in Chinese Pharma-copoeia and some references for the operation standardization of drug mycoplasma test. Methods:Two incubation conditions,namely aerobic conditions and microaerophilic conditions,were compared with respect to the growth status of mycoplasma in liquid and solid media. Results:The growth of mycoplasma was obvious difference between the two incubation conditions,and the microaerophilic con-ditions were better than the aerobic conditions. Conclusion:The microaerophilic conditions can be used in the incubation of mycoplas-ma test,which should be defined and standardized in the future Chinese Pharmacopoeia.

Microbiological data deviations ( MDD) investigation differs from out of the standard ( OOS) investigation in chemical inspection items. Investigating Out-of-Specification( OOS) Test Results for Pharmaceutical Production issued by FDA specifically ex-cludes microbiological and other biological assays. Until recently, there is not a definitive guideline for MDD investigation in the world. Some institutions and experts have explored the practice and theory of MDD investigation. Referred to the existing practical experience and theoretical achievement and based on the provisions of Chinese GMP (2010 revision) and the requirements of Chinese Pharmaco-poeia Ⅱ(2010 edition) Appendix XIX Q, the article discussed the methods for MDD laboratory investigation.

Objective:To investigate the growth-promoting properties and applicability of the mycoplasma test culture media pre-scribed in Chinese pharmacopoeia, and provide reference for the standardization of the drug mycoplasma test method. Methods: Re-spectively using four quantitative detection methods including sensitivity, degree of color change, colony counts and colony diameter, and mycoplasma media widely used in the world as the reference media, the growth-promoting properties of 4 batches of mycoplasma broth and 4 batches of arginine mycoplasma broth from four domestic manufacturers were investigated. Results:The results of sensitivity assay and absorbance detection showed that all the media inoculated with below 100 CFU test microorganisms exhibited visible color change. Furthermore, the results of color change degree and colony diameter showed that there were significant differences among the media products from different manufacturers(P<0. 01). Conclusion:Mycoplasma broth and arginine mycoplasma broth both can sup-port the growth of below 100 CFU test microorganisms. Due to the difference in the growth-promoting properties among the media prod-ucts from different manufacturers, the drug mycoplasma test workers should use more sensitive methods to examine the applicability of the media.

Objective To investigate the cause of metastatic metatarsalgia after hallux valgus surgery and the clinical outcomes of Weil osteotomy for metastatic metatarsalgia after hallux valgus surgery. Methods From July 2009 to Janurary 2012, data of 27 patients (27 feet) with metastatic metatarsalgia of 2nd to 4th head of metatarsal bone after hallux valgus surgery who had been treated by Weil osteotomy were retrospectively analyzed. There were 1 male and 26 females with an average age of 51 years (range, 28-73 years). Metatarsalgia occurred 6-24 months after operation for hallux valgus. 13 feet underwent mini-invasive cervi-cal wedge osteotomy of the first metatarsal, 7 with Chevron procedure, 5 with Akin procedure, and 2 with Lapidus procedure. No shortening in first metatarsal was found in 5 feet with Akin osteotomy, while there were varying degrees of shortening in first meta-tarsal in the remaining 22 feet. The clinical results were evaluated by American Orthopaedic Foot and Ankle Society (AOFAS) and visual analogue scale (VAS) scores. The preoperative and postoperative plantar pressure changes were tested by plantar pressure tester. Results 25 Patients were followed up for 12-42 months (average, 24 months). Among these 25 cases, the metatarsalgia of 23 cases were completely disappeared. The metatarsal plantar lateral metastatic pain occurred in the remaining 2 cases (2 feet) and 1 was relieved by the foot pad, 1 was cured by re-Weil osteotomy. AOFAS score was 46.82 ± 6.13 before surgery and 90.63 ± 1.65 after surgery. The VAS score was 7.5 (6, 7) before surgery and 0.5 (0, 1.0) after surgery. The last follow-up, according to the score of AOFAS toe metatarsophalangeal-interphalangeal joint, of which 23 cases were excellent, 1 good, 1 poor;the excellent and good rate was 96%(24/25). Preoperative pressure under 2nd to 5th metatarsal head were 3.12±1.62 Pa, 5.81±1.92 Pa, 4.63± 2.10 Pa, 3.37±1.57 Pa, 1.67±1.20 Pa and postoperative were 3.33±1.35 Pa, 3.89±1.08 Pa, 3.65±1.96 Pa, 2.25±1.23, (1.48±1.11) Pa. Postoperative pressure under 2nd to 5th metatarsal head were significantly decreased. Conclusion Weil osteotomy can effec-tively adjust the length of the metatarsal and the height of metatarsal head, thus effectively improve the pressure under the metatar-sal head, so it could reach a good effect in the treatment of metastatic metatarsalgia after hallux valgus surgery.

Objective To demonstrate the relationship between Cdc20 mutation and the promotion of colon cancer via Cdc20loxp/+ APCmin/+ villin-cre+/-compound mutant mice.Methods Cdc20loxp/+ APCmin/+ villin-cre+/-compound mutant mice and APCmin/+ mutant mice were generated by mice mating strategy.The colon tumors of two group mice were compared by phenotypic analysis and histology analysis.Results Phenotypic analysis showed that the number of tumors in Cdc20loxp/+ APCmin/+ villin-cre+/-compound mutant mice group and APCmin/+ mutant mice group was 1.2±0.5 and 1.6±0.5, respectively (t =0.215, P =0.588), and the maximum diameter of tumors was (2.7±0.3) cm and (2.5±0.2) cm, respectively (t =0.568, P =0.575).Pathologic type of Cdc20loxp/+ APCmin/+ villin-cre+/-compound mutant mice was adenocarcinoma, while that of APCmin/+ mice was tubular adenoma.Conclusion Cdc20 carrying a null allele can accelerate the promotion of colon cancer in APCmin/+ mice without influence on the tumor number and size.

Objective To observe the effect of surface electromyographic biofeedback (sEMG-BFB) combined with routine swallow training on dysphagic patients with cerebral infarction at recovery stage.Methods Fiftyone dysphagic patients with cerebral infarction were randomly divided into two groups:control group (26 cases) and biofeedback training group (25 cases).The control group was given routine training including orofacial function training,balloon dilatation and behavioral swallowing training,while the biofeedback training group was given behavioral swallowing training was conducted with the guidance of sEMG-BFB in addition to the routine training.Before and after the treatment,videofluoroscopy swallowing study (VFSS) was performed to observe the opening of upper esophageal sphincter (UES).Functional oral intake scale (FOIS) was used to evaluate swallow function.Results Before treatment,there were no significant difference between the two groups in terms of FOIS score and UES opening (P ＞0.05).The FOIS score increased in both groups after treatment (P ＜ 0.05),and the FOIS score was higher in the biofeedback training group than that of the control group (P ＜ 0.05).After treatment,the number of UES complete opening and incomplete opening was 18 and 8,respectively,in the control group,versus 20 and 5,respectively,in the biofeedback training group.UES opening improved in both groups after treatment (P ＜ 0.05).Conclusion Routine swallowing training combined with sEMG-BFB can benefit the dysphagic patients with cerebral infarction for their UES opening and swallowing ability at recovery stage.