Objective To report the clinical results of managing distal tibial fractures with a hybrid external skeletal fixator.Methods From January 2006 to June 2013,39 patients with distal tibia fracture were managed with limited-close or limited-open reduction and a hybrid external skeletal fixator.They were 26 men and 13 women,with an average age of 40.1 years (range,from 23 to 65 years).According to AO classification,15 fractures were of type A3,8 of type B2,10 of type B3,2 of type C2 and 4 of type C3.According to Gustilo classification,of the 12 open fractures,8 were of type Ⅱ,3 of type Ⅲ a and one of type Ⅲ b.According to Tscherne classification of soft tissue injury,4 cases were of grade l,24 of grade 2,and 11 of grade 3.Open fractures were managed first with radical debridement.Those complicated with fibular fracture were managed first with open reduction and internal fixation of distal fibula followed by close or limited-open reduction and minimal internal fixation depending on the position of distal tibial fracture.Next,the hybrid external skeletal fixation was applied.Five cases were immobilized with trans-articular fixators.The data were recorded regarding interval from injury to surgery,operation time,perioperative blood loss,hospital stay,time of external fixation,time of bony union,and complications.The ankle function was evaluated clinically with the Maryland Scale system at the final follow-ups.Results The 39 patients were followed up for 12 to 18 months (average,14.5 months).Primary incision healing was achieved in 37 cases,but the other 2 patients with open fracture suffered delayed wound healing which was cured by dressing changes for 4 weeks.Altogether,38 cases achieved normal fracture union and their average time of external fixation was 13.5 weeks.The time for complete infusion of fracture lines on X-rays averaged 19.7 weeks.Delayed union occurred in one case whose fracture united after removal of the external fixator,internal fixation with a locking plate and autogenous bone grafting.One case was complicated with pin track infection which was healed after debridement,drainage for 8 weeks and removal of the external fixator.No neurovascular complications were observed.According to the Maryland Scale system,the ankle function was excellent in 8 cases,good in 24 and fair in 7,with an excellent and good rate of 82.1％.Conclusions The hybrid external skeletal fixator is good for distal tibial fractures,because it can cffectively protect the skin and minimize invasion to the soft tissues,reducing incidences of skin necrosis and wound infection.Moreover,since it is flexible in screwing and structure formulation,it facilitates wound management,eslpecially in the management of open fractures.

Grape (Vitis vinifera L.) in production is frequently exposed to inadequate light, which significantly affects its agronomic traits via inhibiting their physiological, metabolic and developmental processes. To explore the mechanism how the grape plants respond to the weak light stress, we used 'Yinhong' grape and examined their physiology-biochemistry characteristics and transcriptional profile under different levels of weak light stress. The results showed that grape seedlings upon low intensity shading treatments were not significantly affected. As the shading stress intensity was strengthened, the epidermis cells, palisade tissue, and spongy tissue in the leaves were thinner, the intercellular space between the palisade tissue and spongy tissue was larger compared with that of the control, and the activities of superoxide dismutase, catalase and peroxidase were decreased gradually. Additionally, the soluble protein content increased and the free proline content decreased gradually. Compared with the control, significant changes in plant photosynthetic characteristics and physiology-biochemistry characteristics were observed under high intensity of shading (80%). RNA-seq data showed that the differentially expressed genes between CK and T2, CK and T4, T2 and T4 were 13 913, 13 293 and 14 943, respectively. Most of the enrichment pathways were closely related with the plant's response to stress. Several signaling pathways in response to stress-resistance, e.g. JA/MYC2 pathway and MAPK signal pathway, were activated under weak light stress. The expression level of a variety of genes related to antioxidation (such as polyphenol oxidase and thioredoxin), photosynthesis (such as phytochrome) was altered under weak light stress, indicating that 'Yinhong' grape may activate the antioxidation related pathways to cope with reactive oxygen species (ROS). In addition, it may activate the expression of photosynthetic pigment and light reaction structural protein to maintain the photosynthesis activity. This research may help better understand the relevant physiological response mechanism and facilitate cultivation of grape seedlings under weak light.
Vitis/metabolism* ; Gene Expression Regulation, Plant ; Photosynthesis/genetics* ; Plant Leaves ; Light ; Seedlings/metabolism*

Flavanone 3-hydroxylase (F3H) is a key enzyme in the synthesis of phycocyanidins. In this experiment, the petals of red Rhododendron hybridum Hort. at different developmental stages were used as experimental materials. The R. hybridum flavanone 3-hydroxylase (RhF3H) gene was cloned using reverse transcription PCR (RT-PCR) and rapid-amplification of cDNA ends (RACE) techniques, and bioinformatics analyses were performed. Petal RhF3H gene expression at different developmental stages were analyzed by using quantitative real-time polymerase chain reaction (qRT-PCR). A pET-28a-RhF3H prokaryotic expression vector was constructed for the preparation and purification of RhF3H protein. A pCAMBIA1302-RhF3H overexpression vector was constructed for genetic transformation in Arabidopsis thaliana by Agrobacterium-mediated method. The results showed that the R. hybridum Hort. RhF3H gene is 1 245 bp long, with an open reading frame of 1 092 bp, encoding 363 amino acids. It contains a Fe²⁺ binding motif and a 2-ketoglutarate binding motif of the dioxygenase superfamily. Phylogenetic analysis showed that the R. hybridum RhF3H protein is most closely related to the Vaccinium corymbosum F3H protein. qRT-PCR analysis showed that the expression level of the red R. hybridum RhF3H gene tended to increase and then decrease in the petals at different developmental stages, with the highest expression at middle opening stage. The results of the prokaryotic expression showed that the size of the induced protein of the constructed prokaryotic expression vector pET-28a-RhF3H was about 40 kDa, which was similar to the theoretical value. Transgenic RhF3H Arabidopsis thaliana plants were successfully obtained, and PCR identification and β-glucuronidase (GUS) staining demonstrated that the RhF3H gene was integrated into the genome of A. thaliana plants. qRT-PCR, total flavonoid and anthocyanin contentanalysis showed that RhF3H was significantly higher expressed in the transgenic A. thaliana relative to that of the wild type, and its total flavonoid and anthocyanin content were significantly increased. This study provides a theoretical basis for investigating the function of RhF3H gene, as well as for studying the molecular mechanism of flower color in R. simsiib Planch.
Arabidopsis/metabolism* ; Rhododendron/metabolism* ; Amino Acid Sequence ; Anthocyanins/metabolism* ; Phylogeny ; Flavonoids/metabolism* ; Cloning, Molecular ; Gene Expression Regulation, Plant ; Plant Proteins/metabolism*

Terpene synthase (TPS) plays important roles in the synthesis of terpenoids which are the main fragrances in Rhododendron flowers. To understand the function of TPS genes in terpenoid metabolism in relation to flower aroma formation, we identified all TPS gene family members in Rhododendron by analyzing its genome database. We then used a transcriptomic approach to analyze the differential gene expression patterns of TPS gene family members in the scented flower Rhododendron fortunei compared to the non-scented flower Rhododendron 'Nova Zembla'. The contents of terpenoid compounds in petals of the above two Rhododendron species at different developmental stages were also measured by using qRT-PCR and head space-solid phase micro-extraction combined with gas chromatography-mass spectrometry. Our results showed that a total of 47 RsTPS members, with individual lengths ranged from 591 to 2 634 bp, were identified in the Rhododendron genome. The number of exons in RsTPS gene ranged from 3 to 12, while the length of each protein encoded ranged from 196 to 877 amino acids. Members of the RsTPS family are mainly distributed in the chloroplast and cytoplasm. Phylogenetic analysis showed that RsTPS genes can be clustered into 5 subgroups. Seven gene family members can be functionally annotated as TPS gene family since they were temporally and spatially expressed as shown in the transcriptome data. Notably, TPS1, TPS10, TPS12 and TPS13 in Rhododendron fortunei were expressed highly in flower buds reached the peak in the full blossoming. Correlation analysis between gene expression levels and terpenoid content indicates that the expression levels of TPS1, TPS4, TPS9, TPS10, TPS12 and TPS13 were positively correlated with the content of terpenoids in the petals of R. fortunei at all flower developmental stages, suggesting that these six genes might be involved in the aroma formation in R. fortunei.
Rhododendron/metabolism* ; Phylogeny ; Terpenes/metabolism* ; Family ; Gene Expression Regulation, Plant

Phenylalaninammo-nialyase (PAL) is a key enzyme in the synthesis of methyl benzoate - a plant aroma compound. In order to understand the function of this enzyme in the formation of fragrance in the scented Rhododendron species-Rhododendron fortunei, we cloned a gene encoding this enzyme and subsequently examined the gene expression patterns and the profile of enzyme activity during development in various tissues. The full length of RhPAL gene was cloned by reverse transcription-PCR (RT-PCR) and rapid amplification of cDNA ends (RACE) techniques. The expression levels of RhPAL gene were measured by real-time quantitative reverse transcription PCR (qRT-PCR) and the amount of phenylalanine and cinnamic acid were assayed with LC-MS. The results showed that the ORF sequence of RhPAL gene amplified from the cDNA templates of flower buds had 2 145 bp, encoding 715 amino acids, and shared 90% homology to the PAL amino acid sequences from other species. qRT-PCR analysis showed that the expression of RhPAL in petals during flowering kept in rising even until the flowers wilted. The expression of RhPAL in pistil was much higher than that in stamen, while the expression in the younger leaves was higher than in old leaves. However, the expression level was relatively lower in petal and stamen compared to that in leaves. We also measured the PAL activity by Enzyme-linked immuno sorbent assay in the petals of flowers at different flowering stages. The results showed that PAL activity reached the highest at the bud stage and then decreased gradually to the lowest when the flowers wilted, which followed a similar trend in the emission of the flower fragrance. The phenylalanine and cinnamic acid contents measured by LC-MS were highly correlated to the expression level of RhPAL in various tissues and at different flowering stages, implying that RhPAL plays an important role in the formation of the flower fragrance. This work may facilitate the breeding and improvement of new fragrant Rhododendron cultivars.
Amino Acid Sequence ; Cloning, Molecular ; DNA, Complementary ; Flowers/genetics* ; Rhododendron/genetics*

This study aimed to investigate the effects of Sargassum fusiforme polysaccharide (SFPS I, II, and III) on the apoptosis and regulation of human erythroleukemia (HEL) cells. The effect of different doses of SFPS on HEL cell growth was detected using the Cell Counting Kit-8 method, and apoptosis was detected by Hoechst staining. Cell cycle distribution and apoptosis were detected using flow cytometry. Expression of the cell cycle gene, p53, antiapoptotic genes, Bcl-xL and Bcl-2, and pro-apoptotic genes, Bax, Bad, and Caspase-3, as well as the expression of the corresponding proteins, were detected using real-time quantitative polymerase chain reaction (qPCR) and Western blot. The results showed that SFPS II and III decreased HEL cell viability and induced HEL cell apoptosis. Different concentrations of SFPS (I, II, and III) were detected that induced much less toxic effect in normal human embryonic lung (MRC-5) cells, and SFPS I increased cell proliferation, indicating its favorable selectivity towards cancer cells. The mechanism by which SFPS induced apoptosis was also found to be related to the induction of cell cycle arrest in the G/G phase and the increased expression of apoptosis-related genes and proteins. We concluded that SFPS induces HEL cell apoptosis, possibly via activation of the Caspase pathway, providing the theoretical basis for the development of SFPS-based anti-tumor drug products.

Objective To study the expression of BDNF and TrkB in hippocampal microglia cells of poststroke depression (PSD) rats.Methods Forty healthy adult SD rats were randomly divided into normal group,depression group,stroke group and PSD group (10 in each group).Expression of BDNF and TrkB in OX42-marked hippocampal microglia cells of PSD rats was detected with immunofluorescence staining on day 29 and 57 after a MCAO model rats was established and a chronic stress depression model of rats was established by chronic unpredictable mild stress (CUMS) combined with separate breeding.Results The absorbance value of BDNF and TrkB was 83.35t5.74 and 82.35±5.74 respectively in PSD group on day 29 after the CUMS of rats was established,which was significantly lower than other 3 groups (P＜0.05),and was significantly higher in depression group than in stroke group (141.23±9.16 vs 133.31±7.89;141.23± 8.07 vs 128.62±6.92,P＜0.05).The absorbance value of BDNF and TrkB was 81.63±7.19 and 74.43±7.42 respectively in PSD group on day 57 after the CUMS of rats was established,which was lower than in other 3 groups,and was significantly lower in stroke group than normal group and depression group (93.36±7.56 vs 124.11±11.39、116.65±10.55,87.14±6.56 vs 112.58± 10.99、108.05±10.57,P＜0.05).Conclusion Hippocampal microglia cells play an important role in the pathogenesis of PSD by down-regulating the expression of BDNF and TrkB in PSD rats.

Objective To investigate the association of post-stroke depression (PSD) with tryptophan hydroxylase 2 (TPH2) gene polymorphism and related clinical risk factors of PSD.Methods Three hundred and seventy-six patients diagnosed as having acute ischemic stroke for the first time, admitted to our hospital from March 2015 to September 2017, were chosen in our study. According to Hamilton depression scale scores and Diagnostic and statisistical Manual of Mental Disorders, fourth edition (DSM-IV), these patients were divided into PSD group and non-PSD group. High-resolution melt analysis was used to complete the genotyping ofTPH2gene rs4641528 and rs1386494 in the enrolled patients. The correlations of single nucleotide polymorphisms and PSD were analyzed. Logistic regression analysis was performed on items with statistically significant differences in univariate analysis to identify independent factors affecting PSD.Results There were 104 patients into the PSD group and 272 patients into the non-PSD group; there were statistically significant differences between the two groups in years of education and NIHSS scores (P<0.05). There were no significant differences in rs1386494 genotype and allele frequency between the two groups (P>0.05); there were significant differences in rs4641528 genotype and allele frequency between the two groups (P> 0.05). The C allele inthe chromosomes of PSD patients accounted for 54.3％, while the C allele in the chromosomes of non-PSD patients accounted for 43.4％; the presence of allele (C) increased the risk of PSD (OR=1.552, 95％CI: 1.126-2.141,P=0.007). The results of multivariate Logistic regression analysis showed that baseline NIHSS scores and genotypes of rs4641528 (C/C+C/T) were independent influencing factors of PSD.Conclusion TPH2rs4641528 gene polymorphism and baseline NIHSS scores are found to be associated with depression 3 months after stroke in south Anhui province.

OBJECTIVETo evaluate the sensitivity and specificity of optimized sequential screening program of colorectal cancer, and provide evidence for the further optimization of colorectal cancer screening program.

METHODSUsing cluster sampling method, 4 administrative villages were selected from Jiashan county as a census district in 2011 to 2013. Volunteers of 40 to 74 years old in the census were recruited, and tested by both optimized sequential screening (including questionnaire survey and fecal occult blood test) and colonoscopy for colorectal cancer. Sensitivity and specificity of different screening methods were calculated, respectively.

RESULTSA total of 2 607 volunteers took both simultaneously screening and colonoscopy at the same time. 20 colorectal cancer cases, 85 advanced adenoma cases, 271 non-advanced adenomas cases, and 141 non-adenomatous polyps cases were detected. Sensitivity of optimized sequential screening for colorectal cancer, advanced adenomas, and non-advanced adenomas were 70.0% (14/20) , 57.6% (49/85) and 36.5% (99/271) , specificity was 68.7% (1 776/2 587) , 69.2% (1 746/2 522) and 68.9% (1 610/2 336) , respectively. Sensitivity of the fecal occult blood test of colorectal cancer, advanced adenomas and non-advanced adenomas were 70.0% (14/20) , 47.1% (40/85) and 26.6% (72/271), specificity was 79.4% (2 053/2 587), 79.9% (2 014/2 522) and 79.6% (1 860/2 336). The sensitivity of fecal occult blood test and those of optimized sequential screening for colorectal cancer, advanced adenomas was not significant (χ(2) = 0.00, 1.91, all P values > 0.05). Sensitivity of questionnaire survey of colorectal cancer, advanced adenomas and non-advanced adenomas were 10.0% (2/20), 14.1% (12/85), 12.9% (35/271), specificity was 87.6% (2 266/2 587), 87.7% (2 211/2 522), 87.6% (2 046/2 336). There were no significant difference between non-advanced adenomas. The sensitivity of advanced adenomas and non-advanced adenomas showed no significant decline when the following six term were removed from screening programs: chronic diarrhea, chronic constipation, mucus or bloody history, history of chronic appendicitis or appendectomy surgery, chronic cholecystitis or gallbladder surgery, adverse events in the history of life, while the sensitivity of colorectal cancer remained nearly the same 70.0% (14/20), 52.9% (45/85), 31.4% (85/271) (χ(2) = 0.38, 1.61, all P values > 0.05).

CONCLUSIONCurrent optimized sequential screening programs for colorectal cancer in China have a high sensitivity and specificity. However, further optimization is viable and necessary.

Adenoma ; China ; Colonic Polyps ; Colonoscopy ; Colorectal Neoplasms ; Early Detection of Cancer ; Humans ; Mass Screening ; Occult Blood ; Sensitivity and Specificity ; Surveys and Questionnaires

Objective:To investigate the factors affecting the accuracy of electronic portal imaging device (EPID)-based in vivo dose verification in radiotherapy for patients with lung and esophageal cancers, and to recommend the workflow and specifications for the application of the in vivo dose verification. Methods:This study randomly selected 32 patients who received radiotherapy for esophageal and lung cancers at the Department of Radiation Oncology, Jinhua Municipal Central Hospital from May to August 2022, including 14 lung cancer cases and 18 esophageal cancer cases. Using a uRT-linac 506c linear accelerator, these patients were treated according to the dynamic intensity-modulated radiotherapy (dIMRT) and EPID-based In vivo dose verification ( In vivo EPID) plans developed with the uRT-TPOIS planning system. The In vivo dose verification performed during the treatment included 238 fractions of In vivo EPID and 80 fractions of image-guided radiotherapy (IGRT) for the lung cancer cases, as well as 414 fractions of In vivo EPID and 105 fractions of IGRT for the esophageal cancer cases. The 2D γ passing rate for each irradiation field was obtained according to the set threshold value. Furthermore, fractioned irradiation fields with γ-passing rates below the threshold value were analyzed, and primary factors decreasing the γ-passing rate were further analyzed by combining the online CT images and 3D reconstruction-derived dose. Results:For lung and esophageal cancers, the mean γ-passing rates were 95.1% ± 5.7% and 96.5% ± 4.5%, respectively at 3 mm/5%; 91.5% ± 8.4% and 92.2% ± 4.9%, respectively at 3 mm/3%, and 79.1% ± 14.7% and 83.7% ± 8.2%, respectively at 2 mm/2%, indicating no statistically significant differences between two cancers ( P > 0.05). The average γ passing rate for beam orientations near 0°/180° (Group A) was higher than those near 90°/270° (Group B) 3 mm/5%: Z = -25.4, P < 0.05; 3 mm/3%: Z = -26.8, P < 0.05). The IGRT correction of setup errors significantly improved the γ passing rates (96.3% ± 5.1% and 96.4% ± 4.9%, respectively at 3 mm/5%, Z = -5.50, P < 0.05; 92.3% ± 8.0% and 91.3% ± 7.7%, respectively at 3 mm/3%, Z = -9.54, P < 0.05). The results of In vivo dose verification were affected by changes in the volumes and motion of tumors and normal tissues, radiotherapy positioning, and adequacy of pre-treatment preparation. Conclusions:EPID-based In vivo dose verification during radiotherapy can avoid incorrect irradiation. However, it is necessary to standardize the workflow of the EPID-based In vivo dose verification to avoid the decrease in the γ passing rate caused by artificial factors.