Objective: Based on the data of transcriptome sequencing of Magnolia officinalis, MoDXS1 and MoDXS2 genes were completed in detail by using bioinformatics methods. Methods: MoDXS1 and MoDXS2 genes were analyzed and predicted by the tools of bioinformatics in the following aspects: physical and chemical characteristics of amino acid sequences, function domain, hyophobicity or hydrophilicity, secondary structure and tertiary structure of protein, molecular phylogenetic evolution, and so on; The expression levels of MoDXS1 and MoDXS2 were identified by real-time PCR. Results: ORF Finder indicated that MoDXS1 and MoDXS2 genes were full-length, and they all were unstable hydrophobic proteins; Structural domain of MoDXS1 and MoDXS2 showed high homology with other plants; The secondary structures all were hybrid architecture, and alpha helixes were the major motifs, tertiary structure of protein was predicted by Homology modeling; Sequence alignment that MoDXS family had relative close relationship to the DXS of Nicotiana tabacum, Salvia miltiorrhiza, and Arabidopsis thaliana; The results of evolutionary relationship analysis showed that MoDXS1 had relative close relationship to angiosperm, but MoDXS2 was clustered in a clade solely; The expression levels of DXS1 in M. officinalis and M. officinalis var. biloba were not significantly different, but the expression level of DXS2 in M. officinalis var. biloba was higher than that in M. officinalis. By applying the technique of GC-MS, the contents of major volatile components β-caryophyllene, β-caryophyllene oxide, and β-eudesmol in M. officinalis var. biloba are higher than those in M. officinalis. Conclusion: The results provide theoretical reference for studies on secondary metabolic regulation in terpenoid of M. officinalis.

Twenty-three histone methyltransferase genes were obtained from transcriptome dataset of Lonicera japonica. The nucleotide and proteins characteristics, subcellular localization, senior structural domains and conservative forecasting were analyzed. The result of phylogenetic tree showed that 23 histone methyltransferases were mainly divided into two groups: lysine methyltransferase and arginine methyltransferases. The result of gene expression showed that 23 histone methyltransferases showed preference in terms of interspecies and organs. They were more expressed in buds of L. japonica than in L. japonica var. chinensis and lower in leaves of L. japonica than in L. japonica var. chinensis. Eight genes were specific expressed in flower. These results provided basis for further understanding the function of histone methyltransferase and epigenetic regulation of active ingredients of L. japonica.
Computational Biology ; Gene Expression ; Histone-Lysine N-Methyltransferase ; genetics ; Lonicera ; enzymology ; genetics ; Phylogeny

Methyl valerate (MVA) pathway is one of the important ways for synthesis of terpenoids. This study was based on data of the transcriptome sequencing of Magnolia officinalis, the associated genes MoACOT, MoHMGS, MoHMGR, MoMK in methyl valerate (MVA) pathway, were completed in detail by using bioinformatics methods. The results of analysis showed that MoACOT and MoMK were stable hydrophobic proteins, MoHMGS and MoHMGR were unstable hydrophobic protein. The secondary structures of all proteins were hybrid architecture,and alpha helical were the major motifs. There were no clear transmembrane domains in MoACOT, MoHMGS and MoMK, but two transmembrane domains were founded in MoHMGR which were from 39-61 aa and 82-104 aa resepectively. The results of evolutionary relationship analysis showed that MoACOT, MoHMGS, MoHMGR and MoMK had relative close relationship to angiosperm or dicotyledonous plants, and accorded with genetic evolution rule. From transcriptome data, transcripted level of MoACOT, MoHMGS, MoHMGR, MoMK in M. officinalis and M. officinalis var. biloba was not significantly different. The result provided theoretical reference for study on Methyl valerate (MVA) pathway of terpenoid of M. officinalis.
Computational Biology ; Genes, Plant ; Magnolia ; genetics ; metabolism ; Phylogeny ; Terpenes ; metabolism

The transcriptome represents the whole complement of RNA transcripts in cells or tissues and reflects the expressed genes at various life stages, tissue types, physiological states, and environmental conditions. Transcriptomics study concerning medicinal plants has become the most active area in medicinal plant genome research. Transcriptome analysis provides a comprehensive understanding of gene expression and its regulation. The study of its transcriptome has great significance in solving the questions of genetic evolution, genetic breeding, ecology and so on. Here we report the application status of transcriptomics in medicinal plants based on emergence, development and methodology of transcriptomics.
Gene Expression Regulation, Plant ; Gene Regulatory Networks ; Plants, Medicinal ; genetics ; Sequence Analysis, RNA ; Transcriptome

According to the designed specific primers of gene fragment based on the Salvia miltiorrhiza transcriptome data, a full-length cDNA sequence of SQS2 from S. miltiorrhiza f. alba was cloned by the method of reverse transcription polymerase chain reaction (RT-PCR). The SmSQS2 cDNA sequence was obtained, this sequence is named SmSQS2 and its GenBank registration number is KM244731. The full length of SmSQS2 cDNA was 1245 bp, encoding 414 amino acids including 5'UTR 115 bp and 3'UTR 237 bp. Sequence alignment and phylogenetic analysis demonstrated that SmSQS2 had relative close relationship to the SQS2 of S. miltiorrhiza. The induction of E. coli [pET28-SQS2] in different temperature, induction time, IPTG concentrations and density of inducing host bacterium (A600) were performed, Shaking the culture at 30 degrees C until the A600 is approximately 0.6 and add IPTG to final concentration of 0.2 mmol x L(-1), and then the optimal expression of SmSQS2 recombinant protein were accumulated after the induction time of 20 h. The research provided important base for the study of sterol and terpene biosynthesis of SQS2 in S. miltiorrhiza f. alba.
Cloning, Molecular ; Farnesyl-Diphosphate Farnesyltransferase ; chemistry ; genetics ; metabolism ; Models, Molecular ; Molecular Sequence Data ; Phylogeny ; Plant Proteins ; chemistry ; genetics ; metabolism ; Salvia miltiorrhiza ; chemistry ; classification ; enzymology ; genetics ; Sequence Alignment

Background Prevention and treatment of posterior capsular opacification (PCO) is a hot issue.To establish a PCO animal model is the basis of relevant studies.The most common methods of creating a PCO model are phacoemulsification surgery with or without intraocular lens (IOL) implantation.But the suitability of different methods is unclear.Objective This experiment was to compare the outcome between the two methods of establishing a PCO model in rabbit eyes.Methods Twenty New Zealand white rabbits were collected and randomized into 2 groups.Phacoemulsification of cataract with IOL or without IOL implantation was performed on the right eyes of rabbits in these two groups.The operative eyes were examined under the slit lamp from day 1 through 3 months after surgery.The inflammatory response was evaluated and compared between the two groups,and the extent of PCO was graded based on Odrich's criteria.The use of the animals complied with the Regulations for the Administration of Affairs Concerning Experimental Animals by State Science and Technology Commission.Results The inflammatory response,including conjunctival congestion,corneal edema and aqueous flare were less severe in the model eyes with IOL implantation than the eyes without IOL implantation 1-3 days after operation.Inflammatory response gradually disappeared and showed the same degree in the PCO grade from 2 weeks through 3 months in both groups.The numbers of eyes with 1-3 grade of PCO were 8 and 9,and those with 0 grade of PCO were 2 and 1 in the with IOL implantation group and without IOL implantation group,respectively,showing a significant difference (P=0.39).PCO appeared at 1 month,extended at 2 months and formed dense fibrosis following operation.Conclusions The model outcome of phacoemulsification combined with IOL implantation is better than without IOL implantation one.It is the ideal animal model for the study of after cataract.

AIM:To observe the application efficiency of 5g/L indocyanine green ( ICG ) staining technique for continuous circular capsulorhexis ( CCC ) during phacoemulsification in white cataract.
METHODS:Ninety-eight patients (98 eyes) with white cataract were randomly divided into staining group ( 50 cases, 50 eyes) and control group (48 cases, 48 eyes). The control group didn't do anterior capsule staining. The staining group was injected to fill the anterior chamber, 5g/L ICG 0.1mL was applied on the central surface of the anterior capsule, using a 27G blunt needle through the side-port after 30s, and the redundant ICG was replaced by BSS, and continuous curvilinear capsulorhexis was accomplished using capsulorhexis forceps.
RESULTS: In staining group: after ICG staining, the capsule, which presented uniform light green and visualization of the anterior capsule was significantly improved. There are 48 eyes capsulorhexis success. The rate of success was 96%. Meanwhile, in control group, there was 29 eyes capsulorhexis success. The rate of success was 60%. The difference was statistically significant (P<0. 05). During follow-up, no dye particle and the complications of iritis or high intraocular pressure were found in the eyes of staining group. Anterior chamber inflammation, corneal opacity compared with control group, the difference was not statistically significant.
CONCLUSION:Indocyanine green staining increases the visibility of anterior capsule in over mature cataract, and it should be an effective and helpful method which can increase the success rate of capsulorehxis. At the same time, it can reduce the incidence of intraoperative complications. This will help beginners quickly grasp continuous curvilinear capsulorhexis, and shorten the learning curve.

BACKGROUNDTransplantation of mensenchymal stem cells (MSCs) has been proposed as a promising way for tissue engineering. However, the application of MSCs for transplantation will undergo apoptosis due to the extremely harsh microenvironment such as excessive inflammation. Apigenin (API) has been reported to protect cells against inflammatory damage and cell death by exhibiting anti-inflammatory and anti-oxidative capacity. Here we investigated the modulatory effects of API in lipopolysaccharide (LPS)-mediated inflammation and apoptosis of MSCs, and further defined the underlying mechanism.

METHODSEffects of different concentrations of API (0, 5, 10, 20, 40 and 80 µmol/L) for 24 hours, and LPS (0, 0.5 and 5.0 µg/ml) for 6 hours and 24 hours on MSCs viability were assayed by MTT. Based on this, MSCs were pretreated with different concentrations of API (0 - 40 µmol/L) at the indicated times (6, 12 and 24 hours) followed by exposure to 5 µg/ml LPS for 24 hours. MTT, phase-contrast microscopy, annexinV/propidium iodide (PI) double stain flow cytometry (FCM) and Hoechst staining were applied to explore the effects of API on MSCs induced by 5 µg/ml LPS for 24 hours. In addition, reverse-transcription polymerase chain reaction (RT-PCR) was applied to detect the mRNA expression of pro-inflammatory factors including cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), nuclear factor-kappa B (NF-κB), pro-apoptotic gene caspase-3, Bad, and anti-apoptotic gene Bcl-2. Moreover, AutoDock software was used to imitate the docking score of API and vitamin D receptor (VDR). In parallel, Western blotting and RT-PCR were used to investigate protein and mRNA expression of VDR.

RESULTSMSCs stimulated with LPS 5 µg/ml for 24 hours was used as a model of apoptosis induced by over inflammatory stimulus. API (0 - 40 µmol/L) had non-toxic effect on MSCs; however, it could decrease mRNA expression of COX-2, iNOS and NF-κB at different time points in MSCs induced by LPS, except for API at the concentration of 5 µmol/L.

RESULTSfrom phase-contrast microscopy, MTT, Hoechst staining and AnnexinV/PI double stain FCM demonstrated that with the increasing concentrations of API and extension of administrating time, significant morphological changes of MSCs occurred, viability of cells was strongly inhibited, and meanwhile, apoptosis of LPS-administrated MSCs was exacerbated, compared with LPS individual group. In addition, API promoted caspase-3, Bad mRNA expression and inhibited Bcl-2 mRNA expression in a time-dependent and concentration- dependent manner. Further study found that pro-apoptosis effect of API was related to suppress VDR expression.

CONCLUSIONSAPI could inhibit the expression of inducible inflammatory factors, therefore exert the strong anti-inflammatory function. However, API could not protect MSC apoptosis induced by LPS but amplified the apoptosis. The apoptosis is related to Bad/Bcl-2 increasing and caspase-3 activation, which is mediated through suppressing VDR expression.

Animals ; Apigenin ; pharmacology ; Apoptosis ; drug effects ; Blotting, Western ; Cell Survival ; drug effects ; Cells, Cultured ; Flow Cytometry ; Lipopolysaccharides ; pharmacology ; Male ; Mesenchymal Stromal Cells ; cytology ; drug effects ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptors, Calcitriol ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

Objective To explore the surgical effect to traumatic tentorial herniation with bilater- al mydriasis.Methods The patients were divided into three groups,ie,epidural hematoma group,a- cute diffuse brain swelling group and cerebral contusion and/or subdural hematoma group,to perform clinical outcome analysis.Half year after operation,the neurological outcome was scored according to the Glasgow Outcome Scale.Results Of all,there were three cases with good recovery,10 with moderate disability,nine with severe disability and 10 with vegetative survival but 35 deaths.The outcome was the best in epidural hemotoma group but the poorest in acute diffuse brain swelling group.Conclusions The operative effect of traumatic cerebral herniation with bilateral mydriasis is related with the type of orig- inal injury that is important for selection of operation.Patients with cerebral herniation caused by epidural hematoma should receive immediate operation that will induce better outcome.The operation is not vital for those with cerebral herniation caused by acute diffuse brain swelling.Emergent surgery can save lives of some patients with cerebral contusion and/or subdural hematoma.Rapid diagnosis,correct operation and perioperative treatment may ensure the success of surgery.

"Xishuang" is a special phenomenon that chemical composition of medicinal materials crystallize on the surface exposed to air for a long time. We summarized Herbal textual research of "Xishuang" phenomenon of six herbs, such as Schisandrae Chinensis Fructus, Moutan Cortex, Atractylodis Rhizoma, Magnoliae Officinalis Cortex, dried persimmon frost and watermelon frost. From historical perspective, cream of Schisandrae Chinensis Fructus was firstly discovered in Lei Gong's Moxibustion Theory. Thereafter, dried persimmon frost was found in Song Dynasty, which was named "white persimmon" in Ben Cao Tu Jing and had become an independent medicine in Compendium of Materia Medica. Then, watermelon frost was found in Yang Yi Da Quan of the Qing Dynasty, and Moutan Cortex's "sand star" was recorded in Zeng Ding Wei Yao Tiao Bian of the Republic of China. After that, "Xishuang" phenomenon of Atractylodis Rhizomaand Magnoliae Officinalis Cortex were reported in 1950s and 1960s in succession. The pattern of "Xishuang" is divided into different type, natural "Xishuang" includes Schisandrae Chinensis Fructus, Moutan Cortex, Atractylodis Rhizoma and Magnoliae Officinalis Cortex, artificial "Xishuang" includes watermelon frost, and dried persimmon frost formed crystals by using artificial intervention. The above 6 kinds of herbs have different crystal structure and chemical composition. Therefore, according to traditional identification experience, "Xishuang" phenomenon is related to varieties and quality of medicinal herbs. These research provide herbalism basis for the modern study of "Xishuang" medicinal materials.