Screening of reference genes for quantitative real-time PCR in Artemisia argyi / 中国中药杂志

Artemisia Argyi Folium, a traditional Chinese medicine of important medicinal and economic value, sees increasing demand in medicinal and moxibustion product market. Screening stable and reliable reference genes for quantitative real-time PCR(qRT-PCR) is a prerequisite for the analysis of gene expression in Artemisia argyi. In this study, eight commonly used reference genes, Actin, 18s, EF-1α, GAPDH, SAND, PAL, TUA, and TUB, from the transcriptome of A. argyi, were selected as candidate genes. The expression of each gene in different tissues(roots, stems, and leaves) of A. argyi and in leaves of A. argyi after treatment with methyl jasmonate(MeJA) for different time(0, 4, 8, 12 h) was detected by qRT-PCR. Then, geNorm, NormFinder, BestKeeper, ΔCT, and RefFinder were employed to evaluate their expression stability. The results demonstrated that Actin was the most stable reference gene in different tissues and in leaves treated with MeJA, and coming in the second was SAND. Furthermore, the expression of DXS and MCT which are involved in terpenoid backbone biosynthesis was detected in different tissues and after MeJA treatment. The results showed that the expression patterns of DXS and MCT in different tissues and under MeJA treatment calculated with Actin and SAND as internal reference genes were consistent, which validated the screening results. In conclusion, Actin is the most suitable reference gene for the analysis of gene expression in different tissues of A. argyi and after MeJA treatment. This study provides valuable information for gene expression analysis in A. argyi and lays a foundation for further research on molecular mechanism of quality formation of Artemisia Argyi Folium.

Genome-wide Identification and Characterization of TIFY Gene Family in Medicinal Plant Cannabis sativa / 中国实验方剂学杂志

Objective:The TIFY gene family will be identified and characterized from the whole genome level in Cannabis sativa，which will lay the foundation for gene function study on TIFY family genes and their regulation mechanism on the biosynthesis of cannabinoids and other secondary metabolites. Method:Using the existing genomic data of cannabis，the CsTIFY genes were identified through bioinformatics analysis tools such as NCBI，PlantTFDB，MEME and TBtools etc.，and physicochemical properties，phylogenetic trees，gene structures，chromosome locations and gene expression patterns were analyzed and visualized. Result:Fourteen TIFY family genes（CsTIFY1-CsTIFY14） were identified in Cannabis sativa，which belong to four subfamilies：TIFY，JAZ，ZML，and PPD. The CsTIFYs are composed of 365-1 369 bp nucleotides encoding 118-442 amino acid residues，and their isoelectric points are 4.64-9.96. The 14 CsTIFYs are unevenly distributed on 8 chromosomes，and their proteins are all located in the nucleus. The promoter of CsTIFYs contain multiple abiotic stress responsive cis-acting elements，which indicated that CsTIFYs might involved in the regulation of different abiotic stresses. Transcriptome profiling revealed that CsTIFYs expressed differently in female flowers of 10 differently cannabis varieties，or in flowers，bracts，stems，and leaves of the same variety. Conclusion:Fourteen TIFY family genes were characterized from the whole genome level in C. sativa，and their phylogenetic evolutions and gene expression patterns were analyzed，indicating that CsTIFYs may play important regulatory roles in JA signal transduction，abiotic stress and cannabinoid biosynthesis. This study will provide valuable reference for gene function study of the TIFY family genes in cannabis and cannabis breeding.

MYB Transcription Factors in Fagopyrum tataricum / 中国实验方剂学杂志

Known as "the king of the five grains",tartary buckwheat (Fagopyrum tataricum) is a medicinal and edible plant with both nutritional and healthcare functions. It contains rich flavonoids, such as rutin,with balanced amino acid composition and multiple effects, like blood fat-lowering,blood giucose-lowering,blood pressure-lowering,anti-oxidation,anti-aging,anti-cancer,anti-cancer,and microcirculation-improving. Transcription factors play important roles in plant growth and development by regulating gene expressions. MYB family is one of the largest transcription factor families in plant,contains the MYB domain,and can be divided into four subfamilies:1R-MYB,R2R3-MYB,3R-MYB,4R-MYB. This family plays various roles in plant growth,plant development and flavonoid biosynthesis of secondary metabolism. In this study,the reported MYB transcription factors in F. tataricum were summarized and systemically clustered,and their interrelationships were defined to provide references for further exploring and cloning MYB transcription factors in F. tataricum. In addition,this study reviewed their regulatory functions of MYB transcription factors in flavonoid biosynthesis pathway,plant hormones pathways and other abiotic stress pathways,and made a conclusion and advances about the future research in F. tataricum. Therefore,this study will provide valuable scientific references for the functional studies of MYB family transcription factors in F. tataricum and its molecular breeding for high-quality varieties.

Diagnosis classification and clinical significance of polycystic ovarian syndrome / 中国实用妇科与产科杂志

Polycystic ovary syndrome(PCOS)is char⁃acterized by diverse clinical manifestations and big individual differences.Through the diagnosis classifi⁃cation of PCOS,attention is paid to the different clin⁃ical features,biochemical characteristics and risk of long-term metabolic complications of PCOS patients,which is conducive to the individualized treatment and long-term management of patients.

Identification of 23 unknown Li minority medicinal plants based on DNA barcoding / 中国中药杂志

DNA barcode molecular biological technique is used to identify the species of 23 unknown Li minority medicinal plants.DNA was extracted from 23 unknown medicines using the Plant Genomic DNA Extraction kit. The ITS2 and psbA-trnH regions were amplified and sequenced bi-directionally. The Codon Code Aligner V 7. 0. 1 was used to proofread and assemble the contigs and generated consensus sequences. All the sequences were submitted to Traditional Chinese Medicine DNA Barcode Database and NCBI Gen Bank to get information of the species identifications. If the maximum similarity of the identification result is ≥ 97%,exact species can be known. If it is between 97% and 90%,samples' genus can be confirmed; If it is <90%,then we can only confirm its family. Finally there are 17 samples can be identified to species level,5 can be identified to genus level and 1 can be identified to family level. This shows that DNA barcoding used in medicinal plants molecular identification,can identify unknown species rapidly and accurately.

Molecular genetics research of medicinal plants / 中国中药杂志

With the development of various biotechnology,the research on molecular genetics of medicinal plants has gradually deepened. In this paper,the research system of molecular genetics of medicinal plants was proposed for the first time,which was elaborated from the aspects of genetic resources,genome,gene function and research methods. The application fields of medicinal plant mainly contain species identification,molecular breeding and biosynthesis. The research directions of molecular genetics of medicinal plants in genetic resources,model platform,synthetic biology and molecular breeding were put forward,which include 1 000 genome projects of medicinal plants,model species and mutant libraries,gene original libraries of heterologous synthetic systems,construction gene original library and specific chassis cells in heterologous synthesis system of active ingredient,breeding of new varieties of medicinal plants with high active ingredient and high resistance based on molecular markers andtransgenes.

Application of DNA metabarcoding in species identification of Chinese herbal medicines / 中国中药杂志

DNA metabarcoding,one rapid and robust method using specific standard DNA fragments,has been widely used for rapid species identification of a bulk sample through high-throughput sequencing technologies.While it has been widely used in the studies of metagenomics,animal and plant biodiversity,it has gradually come to be used as a profitable method in species identification of mixed Chinese herbal medicines.In this paper,we mainly summarize the current studies of the application of DNA metabarcoding in species identification of mixed Chinese herbal medicines.Moreover,high-throughput sequencing technologies adopted in those studies,such as Sanger,the next-generation,and third-generation sequencing technologies,are discussed.It is conducted to provide a theoretical guidance for the application of DNA metabarcoding in species identification of mixed Chinese herbal medicines and in more other biodiversity studies.

Statues and research strategy of molecular breeding in Artemisia annua / 中国中药杂志

Malaria is one of the three most deadly diseases in the world. Artemisinin is the first line and effective drug for treating malaria, and only can be extracted from Artemisia annua. Therefore, it is of great significance to cultivate new varieties of A. annua with high artemisinin content. Based on the germplasm bank and the whole genome, transcriptome and genetic map, the authors can explore high-quality genes, stress-resistant genes and genetic markers which have been used for rapid breeding of superior varieties of A. annua. So these methods of molecular breeding will become the main breeding direction of A. annua in the future. The breeding times of new varieties of A. annua can be shortened with molecular breeding technology. Based on the genetic background and the current situation of molecular breeding of A. annua, the strategy and technical route of molecular breeding were discussed and worked out in this paper, which provided a guidance and scientific reference for molecular breeding of A. annua in the future.

Influence of light on gene expression of key synthesis enzyme genes FtANR and FtLAR about proanthocyanidin in seeds of homologous plant of food and medicine Fagopyrum tataricum / 中国中药杂志

Tartary buckwheat Fagopyrum tataricum is an important medicinal and functional herb due to its rich content of flavonoids in the seeds. F.tataricum exhibited good functions for free radicals scavenging， anti-oxidation， anti-aging activities. Although much genetic knowledge of the synthesis， regulation， accumulation of rutin， the genetic basis of proanthocyanidins(PAs) in tartary buckwheat and their related gene expression changes under different lights(blue， red， far red， ultraviolet light) remain largely unexplored. In this study， we cloned one anthocyanidin reductase gene(ANR) and two leucocyanidin reductase gene(LAR) named FtANR，FtLAR1，FtLAR3 involved in formation of(+)-catechin and(-)-epicatechin precusor proanthocyanidin by digging out F. tataricum seed transcriptome data. The expression data showed that the opposite influence of red light on these gene transcript level compared to others lights. The expression levels of FtANR and FtLAR1 decreased and FtLAR3 appeared increment after exposed in the red light， while the expression levels of those genes appeared opposite result after exposed in the blue and far red light.

Effects on recovering of corneal wound and postoperative discomfort of different surgical procedures for pterygium / 国际眼科杂志(Guoji Yanke Zazhi)

?AIM: To evaluate the effects on recovering of corneal wound and postoperative discomfort of different methods for primary pterygium.?METHODS: Forty-seven cases ( 60 eyes ) of primary pterygium were excised under microscope with limbal epithelial transplantation, with sharp dissection ( 24 cases, 30 eyes, Group A) and blunt dissection (23 cases, 30 eyes, Group B).All cases were followed up for 1d to 1mo.?RESULTS: The recovering of corneal wound was better in Group B on 1st day and 3rd day after surgery.Pain, photophobia and tears, foreign body sensation were more serious in group A on 1st day after surgery with a statistically significant difference (P=0.005,0.015,0.012). Pain, photophobia and tears, foreign body sensation were more serious in Group A on 3rd day after surgery with a statistically significant difference ( P=0.019,0.018, 0.015).There was no statistically significant difference on 1wk and 1mo after surgery (P>0.05).? CONCLUSION: Compared with sharp dissection, primary pterygium excised with blunt dissection can significantly improve recovering of corneal wound and postoperative discomfort.

Identification of herbal tea ingredient Mesona chinensis and its adulterants using ITS2 barcode / 中国药学杂志

OBJECTIVE: To establish an accurate and rapid method for identifying Mesona chinensis and its adulterants using the internal transcribed spacer 2 (ITS2) region as a barcode. METHODS: The total genomic DNA from 55 samples representing five species and four variants including M. chinensis and its adulterants were extracted. The ITS2 regions were amplified, and the complete ITS2 sequences were annotated by CodonCode Aligner. The intra- and inter-specific genetic distances based on Kimura 2-Parameter (K2P) and the Neighbor-Joining (NJ) phylogenetic tree were analyzed to identify the species. RESULTS: The lengths of all the 1TS2 sequences of M. chinensis were 232 bp. The intra-specific genetic distances (0) were far shorter than the inter-specific ones between M. chinensis and its adulterants (0.210 2-0.301 9). NJ tree analysis indicated that M. chinensis were accurately differentiated from its adulterants. CONCLUSION: The ITS2 region as an DNA barcode can accurately and effectively distinguish herbal tea ingredient M. chinensis from its adulterants, which provides a quick identifying method to ensure its safe use.

Identification of herbal tea ingredient Plumeria rubra and its adulterants using DNA barcoding / 中国中药杂志

ITS2 sequence was used as a barcode to identify herbal tea ingredient Plumeria rubra and its adulterants. Genomic DNAs from forty eight samples were extracted, the ITS2 sequences were amplified and sequenced bi-direstionlly, and then assembled and obtained using CodonCode Aligner. The sequences were aligned using ClustalW, the genetic distances were computed by kimura 2-parameter (K2P) model and the Neighbor-joining (NJ) phylogenetic trees were constructed using MEGA5.0. Results showed that the length of ITS2 sequence of P. rubra were 244 bp. The intra-specific genetic distances (0-0. 016 6) were much smaller than inter-specific ones between P. rubra and its adulterants(0.320 8-0.650 4). The NJ tree indicated that P. rubra and its adulterants could be distinguished clearly. Therefore, Using ITS2 barcode can accurately andeffectively distinguish herbal tea ingredient P. rubra from its adulterants, which providesa new molecular method to identify P. rubra and ensure its safety in use.

Estimation on the HIV-1 incidence among high risk groups using the pooling RNA technique / 中华流行病学杂志

<p><b>OBJECTIVE</b>To timely identify the HIV-1 infection in window-period and to estimate the HIV-1 incidence among people who came for voluntary counseling and testing (VCT) service as well as men who have sex with men (MSM), respectively.</p><p><b>METHODS</b>HIV antibody negative samples that were determined by screening tests between January and October 2012, were collected and tested with pooling HIV-1 RNA testing technique (2-staged pooling by 50:1, 10:1). Positive cases were followed-up for HIV antibody testing while HIV incidence was calculated under Ron Brookmeyer' s method, among VCT and MSM populations.</p><p><b>RESULTS</b>Among 1400 HIV antibody negative samples of VCT, two showed HIV-1 RNA positive during the antibody window period with the HIV-1 incidence as 1.87% per year (95% CI: 1.23%-2.65% ). Among 500 HIV antibody negative samples from MSM population, two showed HIV-1 RNA positive in the antibody window period, with HIV-1 incidence as 5.31% per year (95% CI: 3.52%-7.45% ).</p><p><b>CONCLUSION</b>Pooling HIV-1 RNA testing seemed a powerful tool for HIV antibody testing in the window-period. Measures should be taken to strengthen the HIV diagnostic programs among MSM and other high risk groups,during the HIV antibody window-period. More frequent detection approach as pooling HIV-1 RNA testing might be a good choice.</p>

Evaluation of PIMA analyzer detecting CD4 cell count of venous and capillary blood in HIV-infected individuals / 中华预防医学杂志

<p><b>OBJECTIVE</b>This study is aimed at evaluating the utility of the portable CD4 analyzers (PIMA).</p><p><b>METHODS</b>The paired finger prick blood (25 µl) and 5 ml venous blood samples were collected from 196 HIV infected patients, who came to Yunnan CDC voluntary counseling and testing (VCT) clinic for CD4 test services, from May to August, 2012. The absolute CD4 cell counts were measured by PIMA (using venous and finger-prick blood) and by Calibur (using venous blood) as the reference. The PIMA and Calibur CD4 results were compared using the Wilcoxon matched-pairs test, and the Spearman's rank correlation coefficients were estimated. The Bland-Altman plots were used to assess the consistency of the two methods.</p><p><b>RESULTS</b>The median absolute CD4 counts of 196 venous blood samples obtained by PIMA and by Calibur were 268 (range:169-403) cells/µl and 302 (range:181-474) cells/µl respectively, which showed significant difference (Z = -7.31, P < 0.01). The median absolute CD4 counts measured by PIMA and by Calibur (using 188 finger-prick and venous blood samples respectively) were 271 (range: 165-450) cells/µl and 304 (range:188-476) cells/µl, which also showed significant difference (Z = -7.60, P < 0.01). The CD4 counts obtained by PIMA CD4 analyzer (using venous and finger-prick blood) showed strong positive correlation with the CD4 counts obtained by the reference method (using venous blood), and the r values were 0.94 and 0.92 respectively (P < 0.01) . The mean biases (limit of agreement) were -38.7 (-210.9-133.5)cells/µl and -45.4 (-221.8-131.0) cells/µl, respectively.Using 350 CD4 counts as the threshold for ART treatment initiation, the sensitivity and specificity of PIMA were 99.1% and 79.3% for venous blood samples, and 97.2%and 78.5% for finger-prick blood samples, respectively.</p><p><b>CONCLUSION</b>The CD4 counts obtained by PIMA are lower than that obtained by Calibur, while the sensitivity is high.</p>

Effect of non-lethal H2O2-induced persistent oxidative injury on retinal pigment epithelial barrier / 中华实验眼科杂志

BackgroundReactive oxygen intermediate products lead to the oxidative injury of cells.Retinal pigment epithelial(RPE) cells produce lots of reactive oxygen intermediate products during the swallow of out disc,but how this procedure cause the persistent oxidative injury of RPE cells is poorly understood.Objective The present study was to evaluate the effect of non-lethal H2 O2 -induced persistent oxidative injury on RPE barrier in vitro.MethodsARPE-19 cell links were inoculated on 96 well plate at the density of 8×104 cells/L and the cell climbing slice of 24 well at the density of 4× 104 cells/L.The cells were cultured in DMEM/F12 medium,and the cells cultured for 24 hours in free-serum medium were used in the experiment.0-0.6 mmol/L of H2O2 were added into the medium.Cellular viability was assessed using 3- ( 4,5-dimethylthiazol-2-yl ) -5- ( 3-carboxymethoxyphenyl ) -2- ( 4-sulfophenyl ) 2H-tetrazolium(MTS) assays.Transepithelial electrical resistance (TER) was used to detect cell monolayer forming time after cultureinTrsnswellchamber.Thepermeabilityof cellmonolayer was examinedbyrhodamine isothiocyanate-dextran transepithelial flux,and immunofluorescence was used to investigate the distribution of the junction protein zonula occludens (ZO-1).ResultsThe total difference was found in the cell vitality(A490) among the different concentrations of H2 O2 ( F =991.501,P =0.000 ).Compared with 0 mmoL/L H2 O2 group,the A490 values was gradually lowed from 0.20 mmol/L H2O2 group to 0.60 mmol/L H2O2 group (P ＜ 0.05 ).H2O2 at the concentrations of ＞0.20 mmol/L lowed the viability of RPE cells.The TER value was ( 24.9 ± 1.3 ) Ω · cm2 in 11 days,( 17.8± 1.4)Ω · cm2 in 7 days after inoculation on transwell chamber,showing a significant difference between them (t=5.228,P=0.014).RPE formed the stable tight junction on day 15 with the TER value (25.9±0.9 ) Ω · cm2.The leakage amount ( relative fluorescence intensity ) of the dextran was 255.39 ± 16.44 in non-H2 O2 control group,exhibiting a significant lowing in comparison with free-cell blank group (433.08±51.53)( t =12.515,P =0.006 ),and that of H2 O2 group was significant increased in comparison with non-H2 O2 control group ( t =14.412,P=0.005).Immunofluyorescence assay showed intact intercellular ZO-1 junction in non-H2O2 control group,but the breakage of ZO-1 junction was seen in H2O2 group.ConclusionsThe results indicate that non-lethal H2O2 can destroy RPE barrier and further lead to the persistent oxidative injury of RPE cells.

A dynamic analysis on incidence and trend of HIV-1 epidemics among Intravenous Drug Users, attendants at the STD clinics and pregnant women in Yunnan province, China / 中华流行病学杂志

Objective BED-CEIA assay was used in HIV/AIDS surveillance sentinel sites to identify recent HIV-1 infection, to estimate HIV-1 incidence and to understand the epidemic trends in Yunnan province. Methods Serum specimens were collected from IDUs in sentinel sites, attendants in STD clinics and pregnant women under a cross sectional study from 2000 to 2007. Specimens confirmed as HIV-1 positive were tested with BED-CEIA to find resent HIV-1 infection, then the annual HIV-1 incidence for each group was calculated and the trends of HIV-1 incidence observed. Results 144 780 serum specimens were collected and 4932 of them were confirmed as HIV-1 positive. 4678 positive specimens were tested with BED-CEIA and 723 ont of them were identified as recent infections. Specimens from the two years were combined for testing. The average HIV-1 prevalence among IDUs was 18.2 %-26.9 % from 2000 to 2007 and the annual incidence were 14.65%, 6.21%, 4.06%, 2.23% respectively. The average HIV-1 prevalence among attendants in STD clinics was 1.6 %-3.2 % and the annual incidence rates were 1.46 %, 0.76 %, 0.52 %, 0.33 % respectively. The average HIV-1 prevalence among pregnant women was 0.2%-0.5% and the annual incidence rates were 0.16%, 0.11%, 0.10%, 0.09% respectively. Conclusion HIV-1incidence rates among IDUs, STDs and pregnant women showed a steady decease.

The analysis of human immunodeficiency virus-1 subtypes in Yunnan province / 中华预防医学杂志

<p><b>OBJECTIVE</b>To analyze the geographical distribution and risk factors of HIV-1 subtypes in Yunnan province.</p><p><b>METHODS</b>Blood samples from 1319 HIV positives were collected in Yunnan Province from 2001 to 2006. The nested polymerase chain reaction was used to amplify the gag (p24)-protease fragments from RNA extracted from plasma or sera. The sequences were used for subtype determination by phylogenetic tree analysis.</p><p><b>RESULTS</b>Among 1319 samples studied, the subtypes has been successfully obtained from 644 samples that were constituted of seven subtypes: CRF08_BC, CRF07_BC, CRF07/08_BC, CRF01_AE, C, B' and URFB/C. C/CRF07_BC/CRF08_BC were distributed in the whole province, but CRF01_AE were mainly distributed in the boarding areas with Myanmar such as Dehong, Baoshan, Xishuangbanna and Puer. Moreover, injecting drugs users accounted for 61.6% (270/438) among C/CRF07_BC/CRF08_BC infections, while only 8.5% (15/177) among CRF01_AE infections.</p><p><b>CONCLUSION</b>Our data indicated that at least seven subtypes were identified in Yunnan province, the relationship between subtypes and transmission routes were analyzed, and the geographic difference of subtypes was also observed.</p>

Observation time for drug administration and withdrawal in the treatment of children with congenital hypothyroidism / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To study the best observation time for drug administration and withdrawal in the treatment of children with transient congenital hypothyroidism,seeking an objective basis for the safe drug withdrawal.</p><p><b>METHODS</b>Levothyroxine was prescribed for 1 144 children diagnosed with congenital hypothyroidism (CH) and according to the results levothyroxine was adjusted to a maintenance dosage. Examinations were performed periodically including physical and mental development, thyroid ultrasonography,and blood levels of T3, T4, TSH. For the patients with a small maintenance dosage of levothyroxine (15.0 - 16.6 g/d) and all the examinations normal, levothyroxine was withdrawn at 2 - 3 years, and the children were followed up and reexamined after 1 month, 2 months, and 10 months, respectively. Permanent drug withdrawal was determined for children with all the examinations normal. Once abnormal TSH occurred, levothyroxine was prescribed again, and followed up continuously.</p><p><b>RESULT</b>Levothyroxine was withdrawn from 157 children. During the follow up, for 15 children (9.55%) levothyroxine were prescribed continuously, and for 142 children permanent drug withdrawal (confirmed with transient CH) was determined. Abnormal TSH of various degrees was detected in 48 cases: 25.48 % (40/157),4.46 % (7/157), and 0.64 % (1/157) were detected at 1, 2 and 10 months after drug withdrawal, respectively. In 15 children levothyroxine was prescribed again for the remarkably high TSH, and the other 33 with mildly abnormal TSH finished the treatment since TSH normalized during follow-up.</p><p><b>CONCLUSION</b>After 2 - 3 years of regular treatment, levothyroxine can be withdrawn from children with normal T3, T4, TSH, physical and mental development, and thyroid function. The best observation time for drug withdrawal should be 2 - 3 months. If T3, T4 and TSH levels are in the normal range, drug can be withdrawn safely. Once abnormal results were detected during follow-up, levothyroxine should be administrated continuously.</p>

Application of CAD/CAM techniques in mandible large-scale defect and reconstruction with vascularized fibular bone graft / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To exploit computer-aided design and computer-aided manufactured (CAD/CAM) techniques and application in the reconstruction of mandible large-scale defect with vascularized fibular bone graft.</p><p><b>METHODS</b>Before actually performing surgery, three-dimensional(3D) computed tomography(CT) was performed in 7 patients with mandibular large-scale defects, and 3D CT images were acquired by processing CT data. Then the CT data were transformed into a readable format and transferred to produce facsimile models by means of using rapid prototyping(RP) techniques. When individual mandibular models and enantiomorphous models were produced, evaluation and surgical simulation was performed in model, which included measuring range of mandible lesions, prefabrication of mandibular reconstructive titanium palate, precise position of titanium screws, shaping the free vascularized fibula by mandibular, etc. According to the simulations, the mandible reconstructions were finished in operation.</p><p><b>RESULT</b>CAD/CAM techniques and application can distinctly display the mandibular lesions and ambient relationships, which is very useful for clinical assessment and surgical planning. Particular advantages were the unlimited trials with the imaging method, and the feeling of reality with the model method. The actual operative time was shortened, and surgery results were satisfactory with few complications.</p><p><b>CONCLUSION</b>CAD/CAM techniques are very helpful for simulation of mandible large-scale defect with complicated anatomical and reconstructive problems. By preoperative simulation of procedures, surgeons can improve or refine treatment planning using this method and improve postoperative results.</p>

Study on the (TAAAA)n repeat polymorphism in sex hormone-binding globulin gene and the SHBG serum levels in putative association with the glucose metabolic status of Chinese patients suffering from polycystic ovarian syndrome in Shandong province / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the relationship of (TAAAA)n repeat polymorphism in the promoter of the sex hormone-binding globulin (SHBG) gene and SHBG serum levels to the glucose metabolic status of Chinese polycystic ovary syndrome (PCOS) patients in Shandong province.</p><p><b>METHODS</b>GeneScan method was used to detect and identify (TAAAA)n repeat number (alleles) and genotypes for 156 controls and 157 patients who were divided into normal glucose tolerance without hyperinsulinemia (NIR group) and with hyperinsulinemia (HI group) and abnormal glucose metabolic (AGM) group according to the results of oral glucose test and insulin resistant test; IRMA was used to measure serum SHBG for part of them.</p><p><b>RESULTS</b>Five alleles containing (TAAAA) 6-10 repeats and 14 genotypes including 6/6, 6/7, 6/8, 6/9, 6/10, 7/7, 7/8, 7/9, 7/10, 8/8, 8/9, 8/10, 9/9, 9/10 repeats genotypes were present in the subjects. Genotype distribution of 6/10 repeats genotype is lower in PCOS than that in control, and 8/9 repeats genotype vice versa (P < 0.01); among PCOS subgroups, the eight repeat genotypes in NIR group is more frequent than that in HI group (P < 0.01), and 7/9 genotype distribution in AGM group is higher than that in NIR group and HI group(P < 0.05-0.01). The serum SHBG levels in homozygous genotype groups exhibit a sequence of 8/8 > 9/9 > 6/6, 7/7 repeats and the fall of serum SHBG trend is in reversed relation with the increase in body mass index (BMI), Homa-IR, and blood pressure. Serum SHBG levels in AGM exhibit a sequence of HI group < NIR group < control but show no statistical difference between both groups.</p><p><b>CONCLUSION</b>This study reveals that the repeat number, alleles, genotypes and their distributions in Chinese women are very different from these in foreigners. Some special genotypes and low serum SHBG levels may be associated with PCOS and its glucose metabolic status; some special genotypes may influence Chinese serum SHBG and need more studies, but both SHBG gene polymorphism genotype and serum SHBG are not good indicators to find out the PCOS individual at high risk.</p>