Background It is well known that sclera remodeling occurs during axial elongation in myopia under the control of growth hormone or its downstream effectors.The role of transforming growth factor-β (TGF-β) in myopia has been determined in previous studies.Bone morphogenetic protein (BMP) is one of members of the TGF-β superfamily,but if it plays an important role in the genesis and development of myopia is not completely clear.Objective This study was to identify the presence of BMPs in normal guinea pigs sclera and investigate the change of BMPs in the sclera in form-deprivation myopia (FDM) of guinea pigs.Methods Thirty young guinea pigs were randomized into normal control group and experimental group using table of random number.FDM models were established by occluding unilateral eyes of guinea pigs with a translucent lens for 14 days in the experimental group,and the fellow eyes served as the controls.Diopter of all eyes was tested by retinoscopy optometry,and ocular axial length was measured by A-sonography before and after modeling.Posterior sclera tissue of the animals was obtained on 14 days,and the relative expression level of BMPs mRNA and protein were assayed by reverse transcription PCR (RT-PCR) and Western blot.The use and care of the animals complied with ARVO Statement.Results On 14 days after occluding of unilateral eyes,the refraction diopter of the experimental group was (-0.48±0.51) D,and that of the fellow eyes was (3.22 ±0.34) D,showing a significant difference between them (t =-12.814,P =0.000).Also,a significant difference in the diopter was seen between the experimental group and normal control group ([-0.48±0.51]D vs.[2.97±0.70]D,t =-11.878,P=0.000).Axial length was (8.30 ± 0.05) mm in the experimental group,(8.11 ±0.06) mm in the fellow eyes and (8.06±0.06) mm in the normal control group,showing a significant increase in the experimental group compared with the fellow eyes and normal control group (t =7.230,P =0.000 ; t =9.084,P=0.000).The expressions of BMP-2 mRNA,BMP-4 mRNA,BMP-5 mRNA in posterior sclera were detected in the normal guinea pigs.Fourteen days after the induction of myopia,the relative levels of BMP-2 mRNA and BMP-5 mRNA in sclera were 0.41 ± 0.11 and 0.65 ± 0.06 in the experimental eyes,which were significantly lower than 0.62 ± 0.07 and 0.84 ± 0.03 in the fellow eyes with the descent range of 34.48％ and 23.67％ respectively (t=2.838,P=0.017; t=2.524,P=0.028).The relative values of BMP-2 protein and BMP-5 protein were 0.44±0.06 and 0.70±0.05 in the experimental eyes,and those of the fellow eyes were 0.61±0.05 and 0.82±0.03,showing significant decline in the experimental eyes with the lowing range of 23.42％ and 15.21％,respectively (t =2.465,P =0.030;t =2.445,P=0.031).No significant differences were found in the expression of BMP-4 mRNA and protein in posterior sclera between the experimental eyes and the normal control eyes (mRNA:t =0.704,P=0.460;protein:t=0.987,P=0.365).Conclusions The expressions of the BMP-2 and BMP-5 in sclera down-regulate significantly in FDM eyes,which suggest that BMP-2 and BMP-5 participate in sclera remodeling during myopia induction.

Objective To investigate the changes of the constitution and its ratio of collagen fiber in the process of masseter reattachment following different osteotomies of the prominent mandibular angle so as to offer guidance for the resection of mandibular angle. Methods Sixteen adult goats were randomized into four groups. In group A we performed unilateral curved osteotomy of the mandibular angle. In group B unilateral curved ostectomy was performed with partial masseter resection. In group C unilateral angle splitting ostectomy, while in group D unilateral dissection of the masseter muscle was conducted. The constitution and its ratio of collagen fiber in the interface were observed at 1-month, 2-month, 3-month, and 6-month after operation. Results On the changes of collagen fiber in the process of muscular reattachment, at 1-month post-operation, the constitution of collagen fiber (types Ⅰ and Ⅲ) in groups A and B were significantly different from that of control group (P<0.05). However, both groups C and D had no statistic difference from control group (P>0.05). At 2-month, 3-month and 6-month post-operation, those of all experimental groups had no statistic difference from control group. And with time, the percentage of collagen fiber type Ⅰ increased and type Ⅲ decreased gradually. Conclusion The recovery sequences of masseter muscle reattachment in this study are firstly group C, secondly group A and finally group B. It suggests that the recoveries of mastication and other oral activities are different. Group B turns out to be with a slow muscle reattachment. Thus, we recommend treating different kinds of mandibular hypertrophy with different ostectomies.

Gentianae Urnulae Herba, dried whole herb of Gentiana urnula,is a commonly used Tibetan medicine. However, only the character identification is used as quality control standard officially at present. As a part of project for the Chinese Pharmacopoeia (2015 edition), the quality standard of this species was established in this study. The tests of water content, total ash, acid-insoluble ash and ethanol-soluble extractives of the crude drugs were carried out following the methods recorded in appendix of Chinese Pharmacopeia (2010 edition, volume 1). The TLC identification method was established by using gentiournoside A as reference substance, and a mixture of ethyl acetate-methanol-water-formic acid(7:1. 5:1: 0. 2) as the developing solvent system on silica gel G TLC plate. The content of gentiournoside A was assayed by HPLC on an Agilent Zorbax SB-C18 (4.6 mm x 250 mm,5 μm) column, using acetonitrile-water (0.1% phosphoric acid) (26:74) as the mobile phase at a flow rate of 1.0 mL x min(-1). The column temperature is at 30 degrees C and the detection wavelength is at 240 nm. As a result, gentiournoside A and the other constituents were separated and presented the same fluorescence light comparing with the reference substance on TLC detected under the UV light(366 nm). The methodology validation for the assay of gentiournoside A showed that it was in a good linear correlation in the range of 0.009 95-0.398 g x L(-1) with the regression equation of Y = 1 467.1X +41.407(r = 0.999 9), and the average recovery was 98. 3% (RSD 2.2%). The mass fractions of gentiournoside A, water content, ethanol-soluble extractives of 15 batches samples were varied in the ranges of 0.175% -1.83%, 8.60% - 9.93% and 29.2% - 35.2%, respectively. Total ash and acid-insoluble ash were 10.2% - 17.2% and 5.26% - 10.8% detected from 10 batches samples. The recommended standards of quantitative indexes are that the mass fractions of gentiournoside A and extractives are not less than 0.80% and 26.0%, respectively; the water, total ash and acid-insoluble ash are not more than 12.0%, 15.0% and 8.0%, respectively.
China ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; standards ; Humans ; Medicine, Tibetan Traditional ; standards ; Plants, Medicinal ; chemistry ; Quality Control

Background The tissue-engineered cornea is becoming the hot spot in the ophthalmologic field,while the research of corneal substitute is in the ascendant,because it is more similar to the corneal morpha and easy to survive in vivo.Objective This study was to investigate the biocompatibility of recombinant human type-Ⅲ collagen/poly9 ( 3-( methacryloylamino ) propyl dimethyl ( 3-sulfopropyl ) ammonium hydroxide ) ( PMPDSAH ) interpenetrating polymer network (IPN) (RHC-Ⅲ/PMPDSAH IPN) hydrogel as a tissue-engineered cornea in rabbit eye and its feasibility as the corneal substitute.Methods One hundred and eight rabbits were randomly divided into experimental group( 90 rabbits) and normal control group ( 3 rabbits),and 15 rabbits ( 30 eyes ) used as the donor corneas.RHC-Ⅲ/PMPDSAH IPN,NGF PMPDSAH IPN and corneal grafts were lamellarly transplanted into the right eyes in RHC-Ⅲ/PMPDSAH IPN group,NGF PMPDSAH IPN group and allograft group respectively.The corneal transparency and neovascularization were examined and scored under the slim lamp and compared among three groups using Kraskal-Wallis H test.The corneal epithelization time was observed and compared among these three groups using one way analysis of variance and LSD-t test.The histological examination of corneas was performed at the 3rd day,1st and 2nd week,1 st,3rd and 6th month after the surgery.The immunohistochemistry was used to detect the expression of K3 in cornea at the 6th month.Results The grafts were well attached in RHC-Ⅲ/PMPDSAH IPN group,NGF PMPDSAH IPN group and allograft group,and no rejection reaction was found throughout 6-month following up.Compared with normal control group,no significant differences were found in the scores of corneal opacification and neovescularization in these three groups (x2 =4.34,P =0.23 ;x2 =2.60,P =0.46 ) at the 6th month.NGF PMPDSAH IPN group achieved reepithelialization in (4.97±0.63) days and was obviously shorted than that in RHC-Ⅲ/PMPDSAH IPN group and allograft group ( t =11.97,P =0.00; t =5.80,P =0.00).The re-epithelialization time in RHC-Ⅲ/PMPDSAH IPN was (6.86±0.71) days,and that of allograft group was (5.87±0.43 ) days,showing a significant difference ( t =6.32,P =0.00).Hematoxylin-eosin staining results demonstrated that implanted materials integrated into the host corneal tissue well and support corneal epithelialization.Part of the material degraded at the 2nd week and degraded completely 1 month later.Regular alignment and distribution of collagen fibers were seen in the regenerated cornea and were similar to those of the normal stroma in 6 months.Immunohistochemistry showed the positive expression of keratin-3 in corneal epithelial cells.Conclusions RHC-Ⅲ/PMPDSAH IPN has a good biocompatibility without toxicity to corneal tissue.Furthermore,NGF can promote the corneal wound-healing and re-epithelialization.The material can be used as safe and reliable corneal substitute after improving the mechanical strength.

Objective To detect the levels of 25-hydroxyvitamin D3 and analyse the related clinical factors in patients with chronic kidney disease (CKD).Methods Patients with CKD in our department from March 1st to July 1st were enrolled continuously.The level of 25-hydroxyvitamin D3 and intact parathyroid hormone(iPTH) were detected by electrochemiluminescence and immunochemiluminescent respectively.Serum calcium,phosphorus and albumin were measured by automatic biochemical instrument.Results 127 patients were selected and the average age was (60.9?15.3).The mean level of 25-hydroxyvitamin D3 was (12.06?6.41)?g/L.82.6% patients had vitamin D deficiency and 96.9% patients had vitamin D insufficiency.The level of 25-hydroxyvitamin had no statistics difference D3 between stage 1,2 and 3 CKD patients but was much hihger than that of stage 4 and non-haemodialysis stage 5 patients.The level of 25-hydroxyvitamin D3 was not related to serum calcium,phosphorus and iPTH,while positively related to albumin and eGFR and negatively related to serum creatinine and total cholesterol.Conclusion Vitamin D deficiency and insufficiency are frequent in CKD patients and deteriorate with the progress of kidney function impairment.The level of 25-hydroxyvitamin D3 is not related to the traditional CKD-MBD index such as serum calcium,phosphorus and iPTH.

Objective:To investigate the effect of metformin and arsenic trioxide on KG1a cells proliferation of acute myeloid leukemia and its possible mechanism.Methods:CCK-8 method was used to detect the killing effect of metformin,arsenic trioxide and combined application on KG1a cells.Annexin V-FITC/P1 Dual Stain Flow Cytometry was used to detect the effect of combined application on apoptosis of KG1 a cells.Western blot was used to detect the expression of intracellular apoptosis-,autophagy-related protein.Results:Metformin and arsenic trioxide alone or in combination could inhibit the proliferation of KG1 a cells and induce apoptosis of KG1 a cells,and the proliferation inhibition rate and apoptosis rate in the combined drug group were higher than those in the drug group alone(P＜0.05).The combination of drugs induced upregulation of Caspase 8 protein and P62 protein expression and was higher than that in the drug group alone(P＜0.05).Conclusion:Metformin can synergize with arsenic trioxide to kill KG1a cells,and its mechanism of action may be related to inducing apoptosis and enhancing autophagy.

The CO I gene sequences of Qianghuoyu, Pachytriton labiatus and Gehyra mutilata were achieved by PCR amplification and bi-directional sequencing. Furthermore, a pair of specific primers SJYW1 and SJYW2 in the non-conservative district were designed through sequence alignment. The PCR reaction condition was established by changing the annealing temperature and cycle numbers. The results showed that 350 bp DNA fragment was amplified from Qianghuoyu in PCR with annealed temperature at 54 °C and the cycle number was 25 cycles, whereas not any DNA fragment was amplified from P. labiatus and G. mutilata under the same reaction condition. This method is well-performed in the identification of Qianghuoyu for its excellent specificity and repeatability.
Animals ; Drug Contamination ; Medicine, Tibetan Traditional ; Polymerase Chain Reaction ; methods

A method for residual determination of 5 pyrethroid pesticides in Anoectochilus roxburghii by cloud point extraction-back extraction-GC-MS was established. PEG 6000 was used as extraction agent and isooctane was used for back-extractant. The con- tent was calculated by external standard method. The linear range was from 15 to 2 000 μg x kg(-1) with the good correlation coefficients (0.955-0.999). The recoveries at spiked concentrations of 50-500 μg x kg(-1) ranged from 85.12% to 101.6%. The limit of detection and quantification of 5 pyrethroid pesticides were in the range of 0.63-3.10 μg x kg(-1) and 2.10-10.31 μg x kg(-1), respectively. The proposed method can be applied to the determination of pyrethroid pesticides residues in A. roxburghii.
Chemical Fractionation ; methods ; Gas Chromatography-Mass Spectrometry ; methods ; Orchidaceae ; chemistry ; Pesticide Residues ; analysis ; chemistry ; isolation & purification ; Pyrethrins ; analysis ; chemistry ; isolation & purification

Objective To study the prescription and preparation technology of breviscapine microemulsion for parenteral injection,and to evaluate its quality.Methods The prescription was selected and optimized through single-factor test and the pseudo-ternary phase diagram method.The preparation technology was investigated,and the particle diameter,drug content,encapsulation efficiency and haemolyticus were evaluated.Results The prescription composition of breviscapine microemulsion was soybean oil:phospholipid:HS15:PEG400:water=1:0.1:0.55:0.55:6.64 (m/m),with the drug content of 4.01 mg·mL-1,the acquired breviscapine microemulsion exhibited light yellow,uniform and transparent,with average particle diameter of 92.1 nm and encapsulation efficiency of 96.8%.The compatibility test showed no drug precipitation and the preparation was no hemolytic crisis.Conclusion The preparation of breviscapine microemulsion injection is correspond to the main index of parenteral injection.

Objective To detect levels of serum platelet activating factor(PAF),thrombomodulin(TM) and white blood cell(WBC),platelet count(PLT) in neonates with meconium aspiration syndrome(MAS),and observe changes of mediators of inflammation and function of endotheliocute.Methods All cases were taken vein blood in 24 h and 72-96 h after birth.Surm PAF and TM were detected by EILSA technique,at the same time,blood cell counts were determined.Results PAF and WBC in neonates with MAS increased,which were relevant to the patients′ condition.TM of neonates with MAS increased significantly,especially in 72-96 h after birth and(aggrava)-ted with the patients′ condition.Conclusion Neonates with MAS have inflammatory reaction and injured endotheliocyte,which are(inte)-raction.