Characterization and phylogenetic analysis of the complete chloroplast genome of Lycopus europaeus / 药学学报

We intend to study the structural characteristics of Lycopus europaeus Linn. chloroplast genome and compare the evolutionary relationship of species from Lamiaceae with similar medicinal effects. The total DNA of Lycopus europaeus was sequenced using the Illumina Hiseq 4000 Sequencing platform and was assembled using NOVOplasty software. And then we annotated and analyzed the genome using the CPGAVAS2 online tool. We constructed the phylogenetic tree using the Stellera chamaejasme and Potentilla chinensis as the outgroup. The whole length of Lycopus europaeus chloroplast genome was 152 085 bp. A total of 132 genes were annotated including 88 protein-coding genes, 8 rRNA genes and 36 tRNA genes. Among them, 8 protein-coding genes (ndhB, rps7, rps12, rps19, rpl2, rpl23, ycf2, ycf15), 7 tRNA coding genes (trnM-CAU, trnL CAA, trnN-GUU, trnE-UUC, trnV-GAC, trnA-UGC, trnR-ACG) and 4 rRNA coding genes (rrn16s, rrn23s, rrn4.5s, rrn5s) are located in the IR region. There are 13 protein coding genes [rps16, rps19 (×2), atpF, rpoC1, rpl2 (×2), petB, petD, rpl16, ndhB (×2), ndhA] each contains one intron, two protein-coding genes (ycf3, clpP) each contain two introns, and 8 tRNA coding genes each contain one intron. A total of 34 SSRs were detected in the chloroplast genome of Lycopus europaeus. Phylogenetic analysis revealed that two species in the Lycopus genus, four species in the Dracocephalum genus, Glechoma longituba, two species in the Mentha genus and Prunella vulgari, in total 10 species are most related. The complete genome sequence of Lycopus europaeus was obtained and analyzed, which clarified the evolutional relationship between the species of Lycopus europaeus and in the Lamiaceae family.

Preparation and in vitro quality evaluation of self-microemulsion co-loaded with tenuifolin and β-asarone / 中国中药杂志

To prepare and optimize the self-microemulsion co-loaded with tenuifolin and β-asarone(TF/ASA-SMEDDS) and evaluate its quality. The prescription compositions of TF/ASA-SMEDDS were screened by solubility test, single factor test and pseudo-tern-ary phase diagram, and the prescriptions were further optimized by Box-Behnken response surface method, with the drug loading and particle size as the evaluation indexes. Then the optimized TF/ASA-SMEDDS was evaluated for emulsified appearance, particle size, morphology and drug release in vitro. The optimized prescription for TF/ASA-SMEDDS was as follows: caprylic citrate triglyceride polyoxyethylene castor oil-glycerol(10.8∶39.2∶50), drug loading of(5.563±0.065) mg·g~(-1) for tenuifolin and(5.526±0.022) mg·g~(-1) for β-asarone; uniform and transparent pan-blue nanoemulsion can be formed after emulsification, with particle size of(28.84±0.44) nm. TEM showed that TF/ASA-SMEDDS can form spherical droplets with a uniform particle size after emulsification; In vitro release test results showed that the drug release rate and cumulative release of tenuifolin and β-asarone were significantly improved. The preparation process of TF/ASA-SMEDDS was simple and can effectively improve in vitro release of tenuifolin and β-asarone.

Comparative analysis of three complete chloroplast genomes of Inula genus with phylogenetic analysis of 49 plants from Carduoideae / 药学学报

italic>Inula japonica, Inula hupehensis and Inula linaariifolia are all medicinal plants of Inula L. in the Compositae family, and Inula hupehensis is endemic to China. In order to compare their genomic sequence differences and provide scientific basis for their germplasm conservation and development, we obtained and analyzed the complete chloroplast genomes of these three species. Total DNAs were extracted from fresh leaves and subjected to next-generation DNA sequencing. NOVOPlasty was used to assemble the chloroplast genomes from the sequence reads. CPGAVAS2 was used to annotate the genes and repeats in each genome. Lastly, phylogenomics analysis was conducted using RAxML. The results showed that the total length of the chloroplast genome of Inula japonica, Inula hupehensis and Inula linaariifolia is 150 754, 150 909, and 150 812 bp respectively, each consisting of a large single copy region, a small single copy region and a pair of inverted repeat regions. In addition, the G/C content of all three chloroplast genomes was approximately 37.7% and each encoded 111 unique genes, including 79 protein-coding, 28 tRNA and 4 rRNA genes. Meanwhile, 32, 33, 34 simple repeat sequences, 18, 22, 18 tandem repeat sequences and 33, 37, 38 scattered repeat sequences were identified in three species. Phylogenomic analysis showed that all three species of Inula L. and Pluchea indica were clustered together, with the relationship between Inuleae and Senecioneae closer, suggesting that Inuleae may have originated from the Senecioneae, not the Cardueae. The data in this study not only enriches the chloroplast genome database of Inula L., but also lays the foundation for the future studies of species identification, phylogenetic relationships, evolution history and genetic diversity of Inula species.

Prevalence and associated factors of intra-articular lesions in acute ankle fractures evaluated by arthroscopy and clinical outcomes with minimum 24-month follow-up / 中华医学杂志(英文版)

Background@#Acute ankle fractures can lead to high rate of concomitant intra-articular lesions which may compromise clinical results. The purpose of this study was to evaluate the Prevalence of concomitant intra-articular lesions in acute ankle fractures with arthroscopy. We also sought to analyze the relationship between intra-articular lesions and the fracture type, as well as the severity of the fracture.@*Methods@#It was a retrospective cohort study. From April 2014 to December 2015, we have chosen arthroscopy-assisted open reduction and internal fixation (AORIF) for the treatment of unstable acute ankle fractures. All concomitant intra-articular lesions were assessed and documented carefully and prospectively, such as ligament injuries, osteochondral lesions, and tibiofibular syndesmosis injuries. All fractures were classified according to the Lauge-Hansen classification system. The American Orthopedic Foot and Ankle Society’s (AOFAS) ankle-hindfoot scale was used to assess post-operative function. Statistical comparisons between the intra-articular lesions, the fracture type, and the severity of the presenting fracture were performed using a Chi-squared analysis.@*Results@#Data of 36 patients were analyzed in the study, including 23 supination-type fractures and 13 pronation-type fractures. The incidence of tibiofibular syndesmosis injuries, chondral lesions, and loose bodies were 92%, 72%, and 39%, respectively. Avulsion fractures of the anterior tibiofibular syndesmosis were more commonly found in supination-type fractures than pronation-type fracture (45% vs. 15%, χ2 = 5.78, P = 0.02), which would cause mechanical blocking in the anterior portion of the ankle. On the contrary, chondral lesions were more commonly found in the more severe fractures than mild fractures (86% vs. 53%, χ2 = 4.57, P = 0.03). A mean 41.7 months (range, 33.0–51.0 months) of follow-up was achieved. A mean AOFAS’s ankle-hindfoot scale was 96.9, and 97.2% of the patients were satisfied with the procedure.@*Conclusions@#Acute ankle fractures have a high incidence of concomitant intra-articular lesions. Avulsion fractures of the anterior tibiofibular syndesmosis are more commonly found in supination-type fractures. Chondral lesions are related to the severity of the fractures, but not with the classification of the fractures. AORIF can be one reliable solution in dealing with the associated injuries seen with acute ankle fractures.

Effect of Traditional Chinese Medicine and Western Medicine Therapy on Gestational Diabetes Mellitus / 中国实验方剂学杂志

In recent years, as the level of economic life has improved, the incidence of gestational diabetes mellitus has increased year by year. Gestational diabetes mellitus (GDM) has been a serious threat to maternal and newborn health. The pathogenesis of gestational diabetes is not very clear, and may be closely associated with insulin resistance, genetic susceptibility, inflammatory response, metabolic disorders. According to the gestational diabetes diagnostic standard,24-28 weeks pregnant women keep an empty stomach over 8 h, taken 75 g oral glucose directly, and then receive the oral glucose tolerance test. GDM is diagnosed as fasting blood-glucose> 5.1 mmol · L-1,1-hour postprandial blood glucose>10.0 mmol · L-1,and 2-hour postprandial blood glucose>8.5 mmol · L-1. Western medicine treatment is mainly based on diet, exercise, drugs, education, monitoring and insulin therapy according to blood glucose. Meanwhile, GDM is a type of diabetes in traditional Chinese medicine. GDM is prevented and treated with diets and traditional method sports and Chinese herbs. Therefore, integrated Chinese and western medicine therapy can maximize the curative effect, reduce the incidence of gestational diabetes mellitus, and effectively improve the adverse outcome and prognosis of patients with gestational diabetes mellitus from mother to child.

Prevalence and associated factors of intra-articular lesions in acute ankle fractures evaluated by arthroscopy and clinical outcomes with minimum 24-month follow-up / 中华医学杂志(英文版)

BACKGROUND@#Acute ankle fractures can lead to high rate of concomitant intra-articular lesions which may compromise clinical results. The purpose of this study was to evaluate the incidence of concomitant intra-articular lesions in acute ankle fractures with arthroscopy. We also sought to analyze the relationship between intra-articular lesions and the fracture type, as well as the severity of the fracture.@*METHODS@#It was a retrospective cohort study. From April 2014 to December 2015, we have chosen arthroscopy-assisted open reduction and internal fixation (AORIF) for the treatment of unstable acute ankle fractures. All concomitant intra-articular lesions were assessed and documented carefully and prospectively, such as ligament injuries, osteochondral lesions, and tibiofibular syndesmosis injuries. All fractures were classified according to the Lauge-Hansen classification system. The American Orthopedic Foot and Ankle Society's (AOFAS) ankle-hindfoot scale was used to assess post-operative function. Statistical comparisons between the intra-articular lesions, the fracture type, and the severity of the presenting fracture were performed using a Chi-squared analysis.@*RESULTS@#Data of 36 patients were analyzed in the study, including 23 supination-type fractures and 13 pronation-type fractures. The incidence of tibiofibular syndesmosis injuries, chondral lesions, and loose bodies were 92%, 72%, and 39%, respectively. Avulsion fractures of the anterior tibiofibular syndesmosis were more commonly found in supination-type fractures than pronation-type fracture (45% vs. 15%, χ = 5.78, P = 0.02), which would cause mechanical blocking in the anterior portion of the ankle. On the contrary, chondral lesions were more commonly found in the more severe fractures than mild fractures (86% vs. 53%, χ = 4.57, P = 0.03). A mean 41.7 months (range, 33.0-51.0 months) of follow-up was achieved. A mean AOFAS's ankle-hindfoot scale was 96.9, and 97.2% of the patients were satisfied with the procedure.@*CONCLUSIONS@#Acute ankle fractures have a high incidence of concomitant intra-articular lesions. Avulsion fractures of the anterior tibiofibular syndesmosis are more commonly found in supination-type fractures. Chondral lesions are related to the severity of the fractures, but not with the classification of the fractures. AORIF can be one reliable solution in dealing with the associated injuries seen with acute ankle fractures.

Complete chloroplast genome of Allium chinense: comparative genomic and phylogenetic analysis / 药学学报

italic>Allium chinense belongs to the genus Alliums of the lily family. It can be used both as medicine and food. To date, the phylogenetic relationship of Allium species have not be resolved completely. Furthermore, there has been a lack of DNA barcode to distinguish closely related species. In this study, the complete chloroplast genome of A. chinense was obtained using next generation DNA sequencing and bioinformatic analysis, and compared with that from other Allium species. The genome is a circular molecule of 152 525 bp with a typical quadripartite structure. Genome annotation identified a total of 116 genes, including 81 protein-coding genes, 31 tRNA genes, and 4 rRNA genes. Analyses of sequences from six Allium species showed that the most diverse regions are found in the protein coding regions such as ndhA and ycf1 genes, and in the intergenic regions, such as ps16-trnQ, trnT-trnF, ndhF-rpl32, rpl32-trnL and rpl16-rps3. A phylogenetic tree was constructed using 58 protein coding sequences from 53 species. All branches showed strong support with bootstrap scores reaching 66%-100%, except those for the Lilium and Paris. Our results suggest that the completed chloroplast genome could solve the classification problems of these species. Using EcoPrimer software, we identified seven markers from the chloroplast genomes, which can be used to differentiate congeneric species. In summary, we have sequenced the complete chloroplast genome of A. chinense, carried out phylogenetic analysis and identified a series of genus specific DNA barcode sequences. The results have laid the foundation for the systematical determination of the phylogenetic relationship of Allium species and the differentiation of species using the genus specific primers.

The influence of implant elastic modulus on stress distributions in implant and peri-implant bone / 医用生物力学

Objective To investigate the effects of changes in elastic modulus of dental implants on stress distributions in implants and peri-implant bone by 3D finite element analysis, so as to supply experimental evidence for new implant system. Methods The model of the mandible with implant bone was constructed based on CT data. The elastic modulus of implants was set as 110, 90, 70, 55 and 40 GPa, respectively. The model was applied with static load of 300 N in vertical direction, 100 N in horizontal direction and 130 N in oblique direction, respectively, to stimulate occlusal state. The stresses on different parts of implants with different elastic modulus and peri-implant bone under 3 kinds of loads were calculated and analyzed. Results As the elastic modulus of implants declined, stresses in cortical bone around implants under horizontal and oblique loads decreased, and stresses in the implants showed a decreasing tendency as well. Conclusions The decrease in elastic modulus of implants can benefit the transferring of load from the implants to the surrounding bone, and reduce the risk of long-term implant failure.

Formulation of dried lignans nanosuspension with high redispersibility to enhance stability, dissolution, and oral bioavailability / 中国天然药物

Herpetospermum caudigerum lignans (HTL), one of the potential drugs with anti-hepatitis B virus and hepatoprotective effects, has limited clinical applications because of poor aqueous solubility and low bioavailability. Both herpetrione (HPE) and herpetin (HPN) are the most abundant ingredients in HTL and exhibit weak acidity. The purpose of the present study was to produce dried preparations of HTL (composed of HPE and HPN) nanosuspensions (HTL-NS) with high redispersibility using lyophilization technology. The HTL-NS was prepared by utilizing precipitation-combined homogenization technology based on acid-base neutralization reactions, and critical formulation and process parameters affecting the characteristics of HTL-NS were optimized. The resultant products were characterized by particle size analysis, SEM, XRD, stability, solubility, dissolution and in vivo bioavailability. HTL-NS showed near-spherical-shaped morphology and the size was 243 nm with a narrow PDI value of 0.187. The dried preparations with a relatively large particle size of 286 nm and a PDI of 0.215 were achieved by using 4% (W/V) mannitol as cryoprotectants, and had a better stability at 4 or 25 °C for 2 months, compared to HTL-NS. In the in vitro test, the dried preparations showed markedly increased solubility and dissolution velocity. Besides, in the in vivo evaluation, it exhibited significant increases in AUC, CMRT and a decrease in T, compared to the raw drug. In conclusion, our results provide a basis for the development of a drug delivery system for poorly water-soluble ingredients with pH-dependent solubility.

Impact of intrauterine Dibutyl phthalate exposure to rat offsprings on reproductive and developmental toxicity / 上海预防医学

Objective ] To observe the effects of intrauterine DBP exposure on the reproductive system of both mother rats and mature F 1 generation rats . [ Methods ] The pregnant rats were administered DBP in various doses (20 mg/kg,100 mg/kg and 500 mg/kg) by gavage.Enzyme linked immunosorbent assay ( ELISA ) was applied to detect the hormone levels in serum , including estrogen two alcohol ( E2 ) , luteinizing hormone ( LH ) , follicle stimulating hormone ( FSH ) and testosterone ( T ) . [ Results] The results showed that the experimental doses of DBP exerted significant effect on serum hormone levels but no significant effects on various organs of mother rats .Compared with the control group , serum T levels in the groups treated with 100 mg/kg and 500 mg/kg were significantly decreased (P<0.05),while FSH level increased in 500 mg/kg dose group (P<0.05);with the increase of DBP exposure doses, the weight of mother rats during gestational period decreased significantly (P<0.05).For the F1 female rats, on PND7, the serum LH and T levels decreased with DBP exposure doses increased (P<0.05);on PND26, the E2 levels of the female pups decreased significantly (P<0.05),the FSH levels of rats in dosed group (100 and 500 mg/kg) were significantly lower than those in the control group (P<0.05),and T level in 500 mg/kg DBP dosed group increased (P<0.05) obviously. [Conclusion] Intrauterine DBP exposure exerts significant toxic effect on the reproductive system of F 1 rats; even low dose of DBP can lead to changes in the serum hormone levels , which can be illustrated by the various metabolic profiles.

Enhancing the accumulation of beta-amyrin in Saccharomyces cerevisiae by co-expression of Glycyrrhiza uralensis squalene synthase 1 and beta-amyrin synthase genes / 药学学报

Glycyrrhiza uralensis Fisch. ex DC is widely used in traditional Chinese medicine (TCM). Among its various active components, glycyrrhizic acid is believed to be the marker component. Squalene synthase (SQS) and beta-amyrin synthase (beta-AS) are key enzymes in the biosynthetic pathway of glycyrrhizic acid in G uralensis. To reveal the effects of co-expression of SQS1 and beta-AS genes on this pathway, 7 yeast expression vectors harboring different SQS1 variants and beta-AS were constructed and expressed in Saccharomyces cerevisiae as fusion proteins. TLC and GC-MS results showed that co-expression of SQS1 and beta-AS enhanced the accumulation of beta-amyrin. The effects of SQS12 were more obvious than the other two SQS1 variants. This study is significant for further investigations concerned with exploring the biosynthesis of glycyrrhizic acid in vitro and strengthening the efficacy of G. uralensis by means of increasing the content of glycyrrhizic acid.

Effects of prior freezing or drying on the swelling behaviour of the bovine cornea / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Frozen or dried corneal grafts are commonly used for stromal transplantation such as lamellar keratoplasty (full or partial thickness), keratophakia, epikeratophakia. Structural properties are important for the final optical results of these surgeries but the effects of freezing/thawing and drying/rehydration on the properties of the stroma are known little compared with the corneal endothelium, mainly because of lack of non-invasive technique to evaluate the stromal structure. This study aimed to investigate the swelling and structural properties of the bovine corneal stroma following freezing or drying by X-ray diffraction which was a non-invasive technique and could give ultra-structural information in hydrated tissues.</p><p><b>METHODS</b>Bovine corneas were either frozen at -40 degrees C or dried to constant weight in a dessicator over silica gel. Swelling was carried out by placing the corneas into dialysis tubing and equilibrating them against various concentrations of polyethylene glycol (PEG) to obtain a range of tissue hydrations. This method minimises the loss of soluble tissue components during the swelling process. Synchrotron X-ray diffraction was used to measure the average intermolecular spacing, the interfibrillar spacing and the fibril diameter as a function of hydration. Changes in light scattering were detected using a microdensitometer.</p><p><b>RESULTS</b>Freezing and thawing of the cornea caused an increase in light scattering by 63.9% at tissue hydration (H) = 3.4, and by 50.0% at H = 4.9. Repeated freezing and thawing causes further increased by 38.9% at the second time and another 36.0% at the third time (P < 0.05). There was a tendency for both the frozen and the dried corneas to lose some swelling ability, achieving hydrations respectively of 10% and 18% below those of fresh corneas at 0 PEG. There were no changes in the fibril diameters, interfibrillar or intermolecular spacings as measured by X-ray diffraction in the equilibrated fresh, pre-frozen and pre-dried corneas.</p><p><b>CONCLUSIONS</b>The increase in light scattering and the loss of swelling ability after freezing and thawing probably results from structural changes following the close association of the collagen molecules and fibrils whilst the tissue is in the dry or frozen state. Some unknown changes in the extracellular matrix between the collagen fibrils may also play a role in the light scattering. The equilibration technique may improve the quality of rehydrated corneal graft or lenticules used for corneal surgeries.</p>

Clinical study of rigid gas-permeable lenses to correct astigmatism for keratoconus / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To evaluate visual outcome and wearing times in eyes fitted with a rigid gas permeable contact lenses and to assess the role of these lenses in therapeutic contact lens practice.METHODS: Patients fitted with RGP from May 2004 to December 2005 were retrospectively reviewed. Ocular diagnosis,flat and steep curvatures by corneal topography. The data included initial base curve, power, and the number of changes made in parameters during the follow-up. The outcome data included visual acuity and the duration of follow-up and lens wear. Complications and complaints were also noted.RESULTS: Forty-Three eyes of 25 patients had keratoconus were reviewed. The mean age of patients was 19.88 ±3.11years. 26 (60.5%)eyes achieved 20/20 visual acuity;15(34.9%) eyes achieved 20/25 to 20/40; and 1 (2.3%) eyes achieved 20/50 to 20/70. 37 (86.0%) eyes achieved visual improvement. No infection or neovascularization was noted; a corneal abrasion occurred in one eye.CONCLUSION: RGP may be a better alternative in selected patients with keratoconus rather than penetrating keratoplasty.

AC OCT of the anterior chamber iris-fixated phakic intraocular lens in high myopia eyes / 国际眼科杂志(Guoji Yanke Zazhi)

· AIM: To study the in situ relative intraocular position of the iris-claw phakic intraocular lens (PIOL)for high myopia using an anterior chamber optical coherence tomography (AC OCT)prototype.· METHODS: Six PIOLs (11.50 to 22.00D lens powers) were implanted in phakic myopic eyes. Using AC OCT, tomography was taken in the anterior chamber to measure the preoperative anterior chamber depth, postoperative distance between the PIOL and the corneal endothelium (endothelial-optic distance), and the postoperative distance between the PIOL and the crystalline lens.· RESULTS: Preoperative anterior chamber depth ranged from 3.27 to 3.91 mm and the postoperative endothelial-optic distance measured 2,07 to 2,24 mm. The distance between the crystalline lens and the posterior surface of the IOL ranged from 0.82 to 1.32 mm. Several tomography revealed the position of the PIOL on the iris, The pigment layer of the iris did not seem to be disturbed by the presence of the PIOL.· CONCLUSION, The original anterior chamber depths were reduced by 36,1% to 44.6% after implantation. This study of 6 eyes revealed that tomography taken by AC OCT are useful in verifying the intraocular position of the PIOL within the anterior chamber. Adequate space was maintained between the iris-fixated phakic intraocular lens and the corneal endothelium, angle, and crystalline lens.

Clinical study of combined penetrating keratoplasty with vitreoretinal surgery / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: to assess the effectiveness of a combined procedure ( pars plana vitrectomy with temporary keratoprosthesis, vitreoretinal surgery, and penetrating keratoplasty). in the complicated cases and the risk factors for the surgical failure.METHODS: Restrospectively reviewed charts of patients who underwent penetrating keratoplasty in combination with vitreoretinal surgery between 1990 and 2005, with a follow-up of 3mo to 9a. Analysis was focused on ocular history, indications for surgery, visual acuity (VA), anatomic results, and complications.RESULTS: 18 eyes had light perception or VA of hand motions only. The best-corrected VA improved during the first 3mo, increased in 72.2% of all eyes, remained unchanged in 27.78%, and no decreased. In 3 of 18 eyes (16.67%), VA was better than finger counting and hand motions, and nine eyes(50%) showed useful vision (0.05) postoperatively. 10 eyes showed a clear corneal graft (55.56%). 2 eyes needs the second keratoplasty, Bullous corneal edema was evident in 3 eyes, band keratopathy was evident in a 3 eyes. 10 patients were observed for more than 2a;6 had a clear graft (60%). Two eyes (11.11%) had silicone oil-corneal endothelium contact and all of these grafts failed.CONCLUSION: Although the functional outcome of a combined procedure is limited by primary and secondary tissue destruction, preserving ambulatory vision is possible and thus improves the quality of life, at least in patients with single remaining eyes.