Secondary metabolite profiles, antimicrobial and antioxidant activities of callus, and leaves extract of Piper sarmentosum Roxb.

This research was aimed to investigate the effect of different combination concentrations of 2,4-diclorophenoxyacetic acid (2,4-D) and benzylaminopurine (BAP) on callus induction, profiles of secondary metabolites, antimicrobial, and antioxidant activities from Piper sarmentosum leaves. These leaves explants were cultured on Murashige and Skoog (MS) medium supplemented with different combination concentrations of 2,4-D and BAP (0.5; 1.0; 1.5; 2.0; 2.5 mg/L) during 6-week culture callus period. Result showed that the morphology of callus grown during this study was compact in various colors appearance, such as light brown, brownish, and dark brown. The fastest mean of callus induction time was 0.5 mg/L 2,4-D and 1.5 mg/L BAP (10 days). The highest callus fresh weight was 1.5 mg/L 2,4-D and 2.0 mg/L BAP (0.350 g). The combination concentration of 1.5 mg/L 2,4-D and 1.5 mg/L BAP was the highest callus dry weight (0.078 g), this treatment was contained 11 different compounds, which were dominated by cyanoacetamide (25.08%) and diisooctyl-phthalate (21.81%). Myristicin and propanamide compounds were also identified in the methanol extract of P. sarmentosum callus. Methanolic extract of callus P. sarmentosum from the optimum treatment had the highest antioxidant activity (26.709 ?L/mL) and antimicrobial activity against three test microbes.

Effect of Media and Photoperiod on Embryogenic and Organogenic Callus Induction from Mature Seed in Oryza sativa L.

Rice is the staple food for billions of people and a source of income for hundreds of millions. This food security crop production needs to be improved to face the projected demands of the exponentially augmenting world’s population and overcome biotic and abiotic stress in the fields. Optimized in vitro culture protocols are a prerequisite to genetic engineering which appears to be the best potential way to improve the rice plant. In this study, ‘Nerica 3’ and ‘Nerica L36’ seeds were in vitro culture tested into 4 media (m1-4), in dark and photoperiod environments. The percentage of callus induction was calculated and callus weight was recorded. Results show that callus induction was influenced by variety × environment and medium × environment interactions, with a strong influence of the environment used. ‘Nerica 3’ showed the highest mean (88.25%) callus induction after six weeks of incubation on different media. m1 and m2 media showed greater mean callus induction (more than 86%). Higher callus induction came from m1 (82% and 91%) and m2 (81% and 93%) media, while lower rates came from m3 (73%) and m4 (69%) media for embryogenic and organogenic calli respectively. Culturing the explant in dark environment to produce embryogenic callus, resulted in greater callus weight for ‘Nerica 3’ (1.03 g) and ‘Nerica L36’ (0.79 g). This study is a contribution to the rice plant genetic improvement by proposing protocols for somatic embryogenesis of two Nerica rice varieties.

Humeral shaft fractures in adults: management and evaluation of treatment in 50 cases

Background: The management of humeral shaft fractures is a real challenge in our regions. Methods: This was a prospective, descriptive single-center study involving 50 patients meeting our inclusion criteria. Inclusion criteria focused on adult patients, received for fractures of the humeral shaft, treated and followed up, then evaluated during the study period. Exclusion criteria included humeral shaft fractures in children, patients not having been evaluated during the study period. Results: We enrolled 50 patients, 43 men (86%) and 7 women (14%), with a sex ratio of 3.07. Mean age was 32.3 years, with extremes of 18 and 79 years. The left side was affected in 46 cases (92%). Lesions were predominantly located in the middle 1/3 in 39 cases (78%). Treatment was orthopedic in 30 patients (60%). Surgical treatment was performed in 20 patients (40%), with screw-plate predominating in 14 cases (28%), followed by hackethal pinning in 5 cases (10%) and combined treatment in one case (2%). At six months' follow-up, our results were very good and good in 36 cases (72%) according to the modified Stewart and Hundley functional score. Conclusions: Humeral shaft fractures are rare fractures for which orthopedic treatment is still indicated. Complications include radial nerve paralysis and pseudarthrosis.

Biomechanical difference of different fixation methods in bone healing of distal tibial comminuted fractures analyzed by finite element method / 中国组织工程研究

BACKGROUND:The treatment of distal tibial comminuted fractures with soft tissue injury has always been challenging.The new retrograde tibial nailing and supercutaneous locking plate are important treatment methods,but their strain and stress shielding at the fracture end during different periods of fracture healing and different load conditions have not been reported. OBJECTIVE:To explore the biomechanical stability of retrograde tibial nailing and supercutaneous locking plate in different periods of fracture healing by finite element analysis to offer a scientific foundation for clinical application and rehabilitation exercise. METHODS:The finite element model of distal tibial comminuted fracture was established by utilizing the CT data of the tibia from a 40-year-old healthy male.Retrograde tibial nailing,supercutaneous locking plate,and callus models were assembled in accordance with the principle of fracture fixation.The finite element analysis was performed using ANSYS software to compare the displacement of the fracture end,the stress shielding of the tibia,the stress of the callus,and the stress distribution of the tibia and the fixation device during different periods of fracture healing. RESULTS AND CONCLUSION:(1)The relative displacement of the tibial fracture decreased gradually with the healing of the fracture,and the displacement decreased significantly after 3 months.At 0 and 1 months after operation,the vertical displacement and total displacement of the supercutaneous locking plate group were higher than those in the retrograde intramedullary nail group.The Z-axis displacement(horizontal medial and lateral displacement)of the two fixation methods was more obvious than the X-axis and Y-axis,and the Z-axis displacement of supercutaneous locking plate group was the most obvious.The maximum Z-axis displacement of the two fixation methods was located on the outside of the tibia,and the minimum displacement was located on the inside of the tibia.(2)The stress shielding rate at different periods of fracture healing gradually decreased with time.The stress shielding rate of the retrograde intramedullary nail was higher than that of the supercutaneous locking plate at different stages of fracture healing.After 3 months,the stress shielding rate of the supercutaneous locking plate was reduced to about 4%,and the stress shielding rate of the retrograde intramedullary nail was reduced to about 40%.(3)The stress of the stress concentration site of the callus in the two fixation methods increased with the increase of the load,and the stress of the callus in the supercutaneous locking plate group was always greater than that in the retrograde intramedullary nail group.The maximum stress distribution of the callus was approximately equally distributed among the two modes of fixation,both in the lateral portion of the tibia.(4)As the fracture healed,the maximum stress of the tibia in the two groups decreased gradually,and the stress in the supercutaneous locking plate group was always greater than that in the retrograde intramedullary nail group.The average stress of the maximum stress area of the tibia in the supercutaneous locking plate group under 1 500 N load was 285 MPa,while that in the retrograde intramedullary nail group was 26 MPa.(5)As the fracture healed,the stress of the fixation device in the two groups decreased gradually,and the stress in the supercutaneous locking plate group was significantly higher than that in the retrograde intramedullary nail group.After 3 months,the stress of the two fixation devices decreased significantly.(6)It is indicated that in the early stage of fracture healing,the strain on the fracture end in the retrograde intramedullary nail group is small,and the maximum stress of the tibia is moderate,allowing early loaded.The fractured ends in the supercutaneous locking plate group had too large strain and too large maximum stress of the tibia,which needed to be partially loaded under protection and could not be fully loaded.In the middle and late stages of fracture healing,the tibial retrograde intramedullary nail and the supercutaneous locking plate could be completely loaded,and the stress shielding rate of the supercutaneous locking plate was significantly lower than the tibial retrograde intramedullary nail.

Thidiazuron-mediated and genotype-independent regeneration system for tomato (Solanum lycopersicum L.)

Solanum lycopersicum L. is one of the most studied crops, with numerous genetic improvement techniques employed to enhance its fruit quality. Recalcitrance of tomato to certain regeneration medium remains to be a limiting factor for a successful in vitro culture protocol. This study was conducted to establish an efficient regeneration system for the target tomato genotype Vb-15 for its use in gene editing using Clustered Regularly Interspaced Short Palindromic Repeats/Cas9 vector system delivered by Agrobacterium tumafaciens. Cotyledons excised from 10 to 12-day-old in vitro germinated seedlings were cultured onto MS medium supplemented with BAP, TDZ, and NAA. Explants cultured on TDZ medium showed enlargement of the explant followed by callusing with multiple shoot development, while explants on BAP-medium showed early signs of callus formation and early development of shoot structures. The highest number of regenerants was obtained in MS with 0.5 mg-1 or 2.5 mg-1 TDZ yielding a mean of 4.45–4.64 shoots per explant. When transferred onto MS basal medium, regenerants from 0.5 mg/L TDZ or BAP showed maximum shoot elongation compared with other treatments. Rooted plantlets transferred to the greenhouse after acclimatization showed 78% survival. The optimum TDZ concentration gave significantly higher regeneration response in seven genotypes compared with Vb 15. Cluster analysis showed that at 0.6 average distances, the response of Vb-15 was unique showing its recalcitrance to in vitro culture.

Differential response of oil palm (Elaeis guineensis Jacq.) genotypes on somatic embryogenesis and plantlet regeneration from zygotic embryo

Oil palm’s enormous economic potential necessitates the genotype evaluation and conservation for its accelerated breeding programs. Hence, in this study, we have evaluated the somatic embryogenesis and regeneration potential of four elite genotypes P-1 (240D X 281D), P-2 (80D X 281D), C-1 (98C X 254D), and C-2 (98C X 208D). For callus induction and proliferation, zygotic embryos (ZEs) were cultured on N6 media supplemented with 2 mgL-1 Dicamba for 90 days. After induction, for somatic embryogenesis and maturation, embryogenic calli were cultured for 120 days in the N6 media with 0.1 mgL-1 2,4-D, 0.16 mgL-1 putrescine, 0.5 mgL-1 casein, and 2.0 g/L activated charcoal. Differentiated polyembryoids were, further, transferred into the regeneration media consisting of N6 with 0.5 mgL-1 NAA, 1.0 mgL-1 BAP, and 0.5 mgL-1 activated charcoal. Among the genotypes studied, P-2 and P-1 have shown the highest callus induction rate, embryogenic calli, differentiated polyembryoids, and more plantlets per somatic embryo cluster. Overall, P-2 and P-1 genotypes have shown conspicuous results over the C-1 and C-2 genotypes in the whole somatic embryogenesis and regeneration process from the matured ZEs of the dura.

Elicitation based enhancement of solasodine production in in-vitro cultures of different Solanum species

Aim: To study the enhancement of solasodine content using elicitors such as NaCl, pectin, salicylic acid and yeast extract in cell suspension cultures of Solanum incanum, Solanum nigrum, Solanum surattense and Solanum villosum. Methodology: In-vitro callus induction from leaf explants was carried out on MS media supplemented with auxin, 2, 4- Dichlorophenoxyacetic acid (2.0 mg l-1). MS liquid medium supplemented with 2, 4-D (2.0 mg l-1) and varied concentrations of different elicitors were used for cell suspension culture. Results: The elicitor NaCl (150 mM) indicated maximum increment in solasodine production in three Solanum species studied. Salicylic acid with 75 ?M resulted in considerable elevation in solasodine content in Solanum spp. Response to elicitation by pectin was high at different concentrations for different species of Solanum. Biotic elicitor yeast extract at 3.0 g l-1 concentration considerably increased solasodine production in S. nigrum and S. villosum. Solanum villosum exhibited best results in terms of solasodine concentration enhancement in response to pectin and yeast extract elicitors whereas Solanum incanum responded best to pectin followed by NaCl. Interpretation: The current results indicated that NaCl, pectin, salicylic acid and yeast extract can be effectively applied as influential elicitors for the enhancing the production of solasodine in cell suspension culture of different Solanum species.

Effect of nutrient media enhanced with plant growth regulators on genetic stability in sub-cultures of Digitalis purpurea callus

This study investigates the effects of nutrient media on the genetic stability of cell lines of callus produced by Digitalis purpurea leaves explants. Four cell lines of D. purpurea callus were selected after inducing 80 sub-cultures of callus in four different nutrient media: Vollosovich (5C01) medium, Murashige and Skoog (MS), Linsmaier and Skoog (LS), and Gamborg (B5) were compared with control samples of seeds and in vitro grown plant (micropropagation). DNA was isolated from the callus by cetyltrimethylammonium bromide, and molecular biomarkers were quantified by inter-simple sequence repeat (ISSR)-polymerase chain reaction (PCR). The purity of the extracted DNA from cell lines and the in vitro has grown plant ranged from 1.94 to 1.98 compared to seeds (1.74) at wavelength 260/280 nm. Out of 360 ISSR-PCR reactions conducted for amplifying 18 samples with 20 primers, 19 primers provided 232 bands with polymorphism and 25 were unique bands with band sizes ranging from 75 to 3500 Bas pair. Cluster analysis and the Jaccard similarity coefficient showed that the medium 5C01 was the most stable compared to the MS, B5, and LS media. The genetic stability of D. purpurea callus was affected by the type and concentration of electrolyte and cytokinin in the nutrient media.

Effect of different plant growth regulators on callus and adventitious shoots induction, polysaccharides accumulation and antioxidant activity of Rhodiola dumulosa / 中草药·英文版

OBJECTIVE@#As a medicinal plant, the resource of Rhodiola dumulosa is deficient along with the large collection. For the protection and utilization of R. dumulosa, the influence of plant growth regulators (PGRs) on callus induction and adventitious shoots differentiation, polysaccharide production and the antioxidant activity were tested.@*METHODS@#Internodes of R. dumulosa were used as explants and cultured on MS medium plus different plant growth regulators (PGRs). The anti-oxidative activities of polysaccharides were evaluated using radical scavenging assays.@*RESULTS@#By response surface plot, 0.85 mg/L N6-benzyladenine (BA), 0.34 mg/L naphthaleneacetic acid (NAA) and 0.33 mg/L 2,4-dicholorophenoxyacetic acid (2,4-D) were the optimal factors for callus induction (90.03%) from internodes explants on MS medium. The fresh weight of green callus increased 47.26 fold, when callus was inoculated on MS + thidiazuron (TDZ) 0.5 mg/L + NAA 2.0 mg/L. Adventitious buds regenerated from callus on the media of MS were fortified with BA 1.0 mg/L plus NAA 0.5 mg/L, and the induction rate was 40.00%. MS plus indole-3-butyric acid (IBA) 1.0 mg/L produced the highest rooting rate with 10 to 15 roots in a length of 2-3 cm per shoot. The content of total polysaccharides in callus developed on MS + TDZ 0.5 mg/L + NAA 2.0 mg/L and MS + BA 1.0 mg/L + NAA 0.5 mg/L was as high as 1.72%-2.15%. At the dose of 0.5 mg/mL polysaccharides extracted from different callus induced on MS + NAA 2.0 mg/L + TDZ 0.5 mg/L or MS + BA 1.0 mg/L + NAA 0.5 mg/L or MS + BA 0.5 mg/L + 2,4-D 0.5 mg/L, the ABTS radical eliminating percentages were 82.78%, 80.18% and 68.59%, respectively, much higher than that of wild plant.@*CONCLUSION@#A rapid micropropagation system for R. dumulosa has been developed. The combination of TDZ and NAA or BA and NAA can increase the yield of the total polysaccharides. The polysaccharides isolated from callus and whole wild plants had stronger free radicals scavenging activities, indicating that polysaccharides from R. dumulosa are the potential pharmaceutical supplements.

Effects of isopsoralen on tibial fracture and vascular healing in mice / 中国骨伤

OBJECTIVE@#To explore effects of isopsoralen (ISO) with different doses on fracture and vascular healing in mice.@*METHODS@#Sixty 2-month-old male C57BL/6 mices with body mass of (20±2) g were selected and divided into 4 groups by random number table method:model group (model), low dose group (isopsoralen-low dose, ISO-L), medium dose group (isopsoralen-medium dose, ISO-M) and high dose group (isopsoralen-high dose, ISO-H), with 15 animals in each group. The right tibial fracture model was established. After operation, ISO-L group, ISO-M group and ISO-H group were given ISO concentration of 10 mg·kg-1, 20 mg·kg-1 and 40 mg·kg-1, respectively. Model group was given same volume of normal saline once a day for 28 days. Weighed once a week. X-ray was performed on 7, 14, 21 and 28 days, respectively, and modified I.R. Garrett scoring method was used to evaluate callus growth. After 28 days, the main organs were stripped and weighed, and organ coefficients were calculated. Hematoxylin eosin staining (HE staining) was performed on the organs to observe whether there were pathological structural changes. Micro-computed tomography (Micro-CT) was used to scan fracture area and conduct three-dimensional reconstruction to obtain the effect map, and quantify bone volume fraction (bone volume/total volume, BV/TV). After decalcification, the tibia was embedded in paraffin wax and sectioned. The healing and shape of fracture end were observed by HE staining and ferruxin solid green staining. The right tibia was removed and decalcified after intravascular infusion of Microfil contrast agent. Micro-CT was used to scan the callus microvessels in the fracture area, and the vascular volume fraction and vessel diameter were quantified.@*RESULTS@#After 28 days of administration, there was no significant difference in body mass and organ coefficient among all groups (P>0.05), and no significant pathological changes were found in HE staining of organs. The results of X-ray and improved I.R. Garrett score showed that ISO-M group was higher than that of Model group at 28 days (P<0.05). Scores of ISO-H group at 14, 21 and 28 days were higher than those of the other 3 groups (P<0.05). Micro-CT results showed intracavitary callus in ISO-M group was significantly reduced, which was lower than that in Model group (P<0.05), most of the callus in ISO-H group were subsided, and BV/TV in ISO-H group was lower than that in the other 3 groups (P<0.05). The results of HE staining and ferrubens solid green staining showed fracture area of ISO-H group was closed, continuous laminar bone had appeared, and the fracture healing process was higher than that of other groups. Angiographic results showed vascular volume fraction in ISO-H and ISO-M groups was higher than that in Model and ISO-L groups (P<0.05), and the vascular diameter in ISO-H and ISO-M groups was higher than that in Model and ISO-L groups (P<0.05).@*CONCLUSION@#In the concentration range of 10-40 mg·kg-1, ISO has no obvious toxic and side effects, and could improve bone microstructure, promote formation of callus microvessels, and accelerate healing of fracture ends in a concentration-dependent manner.

Cloning and expression analysis of U6 promoters in Panax quinquefolius / 中国中药杂志

The U6 promoter is an important element driving sgRNA transcription in the CRISPR/Cas9 system. Seven PqU6 promo-ter sequences were cloned from the gDNA of Panax quinquefolium, and the transcriptional activation ability of the seven promoters was studied. In this study, seven PqU6 promoter sequences with a length of about 1 300 bp were cloned from the adventitious roots of P. quinquefolium cultivated for 5 weeks. Bioinformatics tools were used to analyze the sequence characteristics of PqU6 promoters, and the fusion expression vectors of GUS gene driven by PqU6-P were constructed. Tobacco leaves were transformed by Agrobacterium tumefaciens-mediated method for activity detection. The seven PqU6 promoters were truncated from the 5'-end to reach 283, 287, 279, 289, 295, 289, and 283 bp, respectively. The vectors for detection of promoter activity were constructed with GUS as a reported gene and used to transform P. quinquefolium callus and tobacco leaves. The results showed that seven PqU6 promoter sequences(PqU6-1P to PqU6-7P) were cloned from the gDNA of P. quinquefolium, with the length ranged from 1 246 bp to 1 308 bp. Sequence comparison results showed that the seven PqU6 promoter sequences and the AtU6-P promoter all had USE and TATA boxes, which are essential elements affecting the transcriptional activity of the U6 promoter. The results of GUS staining and enzyme activity test showed that all the seven PqU6 promoters had transcriptional activity. The PqU6-7P with a length of 1 269 bp had the highest transcriptional activity, 1.31 times that of the positive control P-35S. When the seven PqU6 promoters were truncated from the 5'-end(PqU6-1PA to PqU6-7PA), their transcriptional activities were different in tobacco leaves and P. quinquefolium callus. The transcriptional activity of PqU6-7PA promoter(283 bp) was 1.59 times that of AtU6-P promoter(292 bp) when the recipient material was P. quinquefolium callus. The findings provide more ideal endogenous U6 promoters for CRISPR/Cas9 technology in ginseng and other medicinal plants.

Effect of BBM gene on callus growth and ginsenoside content in Panax quinquefolius / 中国中药杂志

Baby Boom(BBM) gene is a key regulatory factor in embryonic development and regeneration, cell proliferation, callus growth, and differentiation promotion. Since the genetic transformation system of Panax quinquefolius is unstable with low efficiency and long period, this study attempted to transfer BBM gene of Zea mays to P. quinquefolius callus by gene gunship to investigate its effect on the callus growth and ginsenoside content, laying a foundation for establishing efficient genetic transformation system of P. quinquefolius. Four transgenic callus of P. quinquefolius with different transformation events were obtained by screening for glufosinate ammonium resistance and molecular identification by PCR. The growth state and growth rate of wild-type and transgenic callus were compared in the same growth period. The content of ginsenoside in transgenic callus was determined by ultra-high performance liquid chromatography-triple quadrupole mass spectrometry(UPLC-MS/MS). The results showed that transgenic callus growth rate was significantly higher than that of wild-type callus. In addition, the content of ginsenoside Rb_1, Rg_1, Ro, and Re was significantly higher than that in wild-type callus. The paper preliminarily proved the function of BBM gene in promoting growth rate and increasing ginsenoside content, which provided a scientific basis to establish a stable and efficient genetic transformation system for Panax plants in the future.

Treatment strategy for pediatric supracondylar humeral fractures with callus formation and displacement neglected for over 1 week / 中华创伤骨科杂志

Objective:To investigate the treatment strategy for pediatric humeral supracondylar fractures with callus formation and displacement neglected for over 1 week.Methods:A retrospective analysis was made of the clinical data of 36 children who had been treated at Department of Pediatric Orthopaedics, Medical Center, The Second Affiliated Hospital, Inner Mongolia Medical University from January 2011 to January 2021 for humeral supracondylar fractures with callus formation and displacement neglected for over 1 week. There were 22 boys and 14 girls, with an age of (6.7±2.7) years (from 2.3 to 12.8 years). All fractures were Gartland type Ⅲ. The patients were divided into 2 groups according to their treatment methods: a closed reduction and percutaneous pinning (CRPP) group of 15 patients subjected to the CRPP treatment only, and a leverage group of 21 patients subjected to CRPP assisted by the "lever technique" with posterior elbow Kirschner wire prying and pulling. The 2 groups were compared in terms of operation time, fluoroscopy frequency, quality of reduction, and recovery time for elbow range of motion; the elbow range of motion, visual analogue scale (VAS), Mayo elbow performance score (MEPS) and complications were assessed at the last follow-up.Results:The 2 groups were comparable because there was no significant difference between them in the general information before operation ( P>0.05). All patients were followed up for (26.2±16.3) months (from 6 to 96 months). All the fractures obtained acceptable reduction and clinical union 4 to 6 weeks after operation. The operation time [(28.2±6.8) min] and fluoroscopy frequency [(27.0±6.0) times] in the leverage group were significantly less than those in the CRPP group [(40.8±10.8) min and (43.3±11.4) times] ( P<0.05). The CRPP group was significantly better than the leverage group in the intraoperative Baumann angle (78.1°±1.6° versus 73.7°±4.1°), lateral capitellohumeral angle (58.3°±2.6° versus 49.6°±5.2°) and horizontal rotation rate (109.5%±3.0% versus 103.2%±4.9%) ( P<0.05). The intraoperative reduction in the CRPP group was significantly closer to the normal mean value than that in the leverage group ( P<0.05). There was no significant difference in the recovery time for elbow range of motion between the CRPP and the leverage groups ( P>0.05). At the last follow-up, the Baumann angle (75.4°±2.8°) and the lateral capitellohumeral angle (53.2°±3.6°) in the leverage group were still significantly better than those in the CRPP group (78.3°±1.5° and 57.5°±2.3°) ( P<0.05). However, there was no significant difference in the elbow range of motion, VAS, MEPS or incidence of complications between the 2 groups ( P>0.05). Conclusion:To treat humeral supracondylar fractures with callus formation and displacement neglected for over 1 week in children, CRPP assisted by the "lever technique" with posterior elbow Kirschner wire prying and pulling is an efficient and accurate method, because it can lead to more satisfactory reduction than CRPP only.

Application of tissue culture technology of medicinal plants in sustainable development of Chinese medicinal resources / 中国中药杂志

Chinese medicinal resources are the cornerstone of the sustainable development of traditional Chinese medicine industry. However, due to the fecundity of species, over-exploitation, and limitations of artificial cultivation, some medicinal plants are depleted and even endangered. Tissue culture, a breakthrough technology in the breeding of traditional Chinese medicinal materials, is not limited by time and space, and can allow the production on an annual basis, which plays an important role in the protection of Chinese medicinal resources. The present study reviewed the applications of tissue culture of medicinal plants in the field of Chinese medicinal resources, including rapid propagation of medicinal plant seedlings, breeding of novel high-yield and high-quality cultivars, construction of a genetic transformation system, and production of secondary metabolites. Meanwhile, the current challenges and suggestions for the future development of this field were also proposed.

Evolución de la osteosíntesis de los huesos largos: fijación por enclavado intramedular con clavos rígidos

Se presenta la evolución histórica y científica de la osteosíntesis de huesos largos realizada con clavos rígidos. Mediante una revisión bibliográfica retrospectiva en revistas de Traumatología nacionales y extranjeras, así como libros relevantes de esta especialidad, se pudieron establecer tres momentos claves en la osteosíntesis intramedular con clavos rígidos y caracterizar las diferentes generaciones que aportaron a este procedimiento en la historia de la Ortopedia y la Traumatología. Destacan las innovaciones tecnológicas incorporadas a la práctica quirúrgica y el desarrollo de biomateriales para mejorar la reparación de lesiones e incorporar al paciente a su vida cotidiana. El trabajo demuestra cómo ha avanzado la técnica quirúrgica de fijación intramedular y la consolidación ósea; y gracias a ello los problemas de las fracturas han quedado prácticamente solucionados.

The historical and scientific evolution of osteosynthesis of long bones performed with rigid nails is presented. Through a retrospective bibliographic review in national and foreign Traumatology journals, as well as relevant books of this specialty, it was possible to establish three key moments in intramedullary osteosynthesis with rigid nails and characterize the different generations that contributed to this procedure in the history of Orthopedics and Traumatology. The technological innovations incorporated into surgical practice and the development of biomaterials to improve the repair of injuries and incorporate the patient into their daily lives stand out. The work demonstrates how the surgical technique of intramedullary fixation and bone consolidation has advanced; and how thanks to this the problems of fractures have been practically solved.

Ilex paraguariensis: the effect of genotypes and growth phase on biomass, secondary metabolism and antioxidant activity of in vitro cultivated calli / Ilex paraguariensis: el efecto de los genotipos y la fase de crecimiento sobre la biomasa, metabolismo secundario y la actividad antioxidante de los callos cultivados in vitro

Yerba mate ( Ilex paraguariensis ) produces several secondary metabolites of interest to the phar maceutical industry, such as chlorogenic acids and methylxanthines. These compounds have been produced in vitro by callus culture from different species. However, for I. paraguariensis , no studies upon the production of these compounds in vitro have been p erformed to date. In this work, we show that the concentration of secondary metabolites from I. paraguariensis callus is possible and highly dependent on the callus growth phase. We observed that the best phase for the production of secondary compounds in calli of yerba mate is the stationary growth phase on both genotypes tested. In this phase, higher levels of phenolic compounds, chlorogenic acid and 3,5 - dicaffeoylquinic acid and greater antioxidant activity were observed. Chlorogenic acid and 3,5 - dicaffe oylquinic acid presented positive correlation with antioxidant activity. For the first time, secondary compounds were reported in yerba mate calli cultivated in vitro .

La yerba mate ( Ilex paraguariensis ) produce varios metabolitos secundarios de interés para la industria farmacéutica, como los ácidos clorogénicos y las metilxantinas. Estos compuestos se han producido in vitro mediante cultivo de ca llos de diferentes especies. Sin embargo, para I. paraguariensis , hasta la fecha no se han realizado estudios sobre la producción de estos compuestos in vitro . En este trabajo, mostramos que la concentración de metabolitos secundarios desde callos de I. pa raguariensis es posible y altamente dependiente de la fase de crecimiento del callo. Observamos que la mejor fase para la producción de compuestos secundarios en callos de yerba mate es la fase de crecimiento estacionario en ambos genotipos probados. En es ta fase se observaron niveles más altos de compuestos fenólicos, ácido clorogénico y ácido 3,5 - dicafeoilquínico y una mayor actividad antioxidante. El ácido clorogénico y el ácido 3,5 - dicafeoilquínico presentaron correlación positiva con la actividad antio xidante. Por primera vez, se reportaron compuestos secundarios en callos de yerba mate cultivados in vitro .

Induction of somatic embryogenesis in Tecomella undulata (Sm.) Seem using a pistillate explant

Desert teak (Tecomella undulata (Sm.) Seem) a multipurpose ornamental tree native to the arid and semi-arid tropics has entered the endangered plant category mainly as a result of the species' ineffective seed reproduction system. The tree usually reproduces through a few root suckers in old stands. Conventional methods of plants multiplication could not offer a viable practice for its mass multiplication. Low adventitious rooting of the cuttings has been the principal cause of failure in its vegetative propagation. Hence, the present research was planned and conducted to evaluate the feasibility of somatic embryogenesis in this species, the pathway that bypasses the need for rooting stage by developing bipolar embryos. Ovary explant was cultured in modified Murashigue & Skoog (MS) medium supplemented with different auxins and cytokinins. The results showed ?-naphthalene acetic acid (NAA) was superior in inducing embryogenic callus. NAA ranging 5.4-21.5 ?M could induce the highest embryogenic calli which exhibited developing pro-embryogenic masses (PEM) and globular somatic embryos. The calli which were induced by the use of 2,4-dichlorophenoxyacetic acid (2,4-D) were poor in quality and showed no morphogenesis potency. Individual application of Thidiazuron (TDZ) and N6-benzyladenine (BA) induced good callogenesis at low concentrations but the calli were non-embryogenic with both. The proliferation of embryogenic calli was the best in a hormone-free medium. However, the media containing 40.5 and 54 ?M NAA alone could induce somatic embryos along with callus proliferation. Low BA-contained medium (0.9-4.44 ?M) led to recurrent somatic embryogenesis. Neither BA and GA3, nor the elevating sucrose concentration could cause further development and maturation of the somatic embryos induced during previous stages (callogenesis and callus proliferation). More research is required to optimize the maturation stage. The findings of the present study can be useful for future studies in the micropropagation of this recalcitrant specieslationships in Indian mustard under heat stress and the differential remobilization efficiencies in the advanced breeding lines.

Effect of local gene therapy on expression of Wnt3a and β-catenin in callus of distraction gap during mandibular distraction osteogenesis in rabbits / 中华医学美学美容杂志

Objective:To investigate the effect of electroporation-mediated local gene therapy on the expression of Wnt3a and β-catenin in callus of distraction gap during mandibular distraction osteogenesis of rabbits.Methods:The experiments were conducted in the laboratory of the Affiliated Friendship Plastic Surgery Hospital of Nanjing Medical University from September 2019 to December 2019. Forty eight New-Zealand rabbits were randomly divided into control group (group A), gene therapy group (group B) and normal saline group (group C), with 16 rabbits in each group. After bilateral mandible osteotomy and distractors were implanted, the distractors were activated at a speed of 0.8 mm/d on 4th day, postoperatively, and lasted for 7 days, followed by consolidation period. Group A distracted only, group B was subject to local injection of recombinant plasmid pIRES-hBMP2-hVEGF165 in the distraction gap and electroporation stimulation at the beginning of activation distractors; and group C local injection of the same dose of normal saline in the distraction gap and electroporation stimulation at the beginning of activation distractors. Four animals in each group were sacrificed on the day at the end of distraction, 7th, 14th, 28th days of consolidation period, respectively. The callus in the distraction gap was taken for immunohistochemical staining and RT-PCR to detect the expression of Wnt3a and β-catenin, and image analysis was performed. SPSS 22.0 statistical software was used for data analysis.Results:Immunohistochemical staining showed that Wnt3a and β-catenin were mainly located in the cytoplasm and nuclei of fibroblasts, chondrocytes and osteoblasts in callus tissue. Immunohistochemistry and RT-PCR showed that the expression of Wnt3a and β-catenin reached a peak at the end of distraction. With the disappearance of distraction tension, the expression of Wnt3a and β-catenin gradually decreased. After gene therapy intervention, the expression of Wnt3a and β-catenin was significantly increased, and the expression of Wnt3a and β-catenin in group B was the highest at each time point, with statistically significant difference compared with groups A and C ( F=96.3, P<0.01). Conclusions:Gene therapy promotes the expression of Wnt3a and β-catenin in the callus of distraction gap, regulating the balance of the bone reconstruction system and thus promoting the formation of new bone in the distraction gap.

Monitoring the Progress of Treatment in Fracture Non-Union: The Role of Alkaline Phosphatase and Ultrasonography / Monitoramento da evolução do tratamento da ausência de consolidação da fratura: Papel da fosfatase alcalina e da ultrassonografia

Abstract Objective To evaluate the role of serum alkaline phosphatase (ALP) and ultrasonography (USG) in monitoring the progress of treatment in diaphyseal non-unions. Methods This prospective observational cohort study included adult patients with diaphyseal fractures of major long bones previously treated with internal fixation and eventually resulting in non-union. Following the definitive treatment for non-union, the patients were followed-up periodically for six months, and serial monitoring of the levels of ALP and USG were performed along with radiographs (X-rays) to ascertain the status of the union. Results After an initial rise at seven weeks, ALP levels declined to normal values in fractures which united, whereas they remained high in cases of persistent non-union. Similarly, after an elevation of the vascular resistive index (RI) at around 12 weeks in all the patients, it decreased in cases progressing to union, while it remained persistently high even at 24 weeks in fractures failing to unite. Cases of persistent non-union continued to show hypoechogenic callus at 24 weeks instead of converting into hyperechogenic callus, as observed in cases which progressed to union. Conclusion Significant changes suggestive of union appeared simultaneously on the X-rays, USG and ALP levels during the follow-up. However, a serial examination of the ALP levels and USG during the follow-up gave a hint of the direction of progress in the healing process of fracture non-union. Their role in monitoring the outcome of nonunion is more complimentary than supplementary to the X-rays.

Resumo Objetivo Avaliar o papel da concentração sérica de fosfatase alcalina (FA) e da ultrassonografia no monitoramento do progresso do tratamento da ausência de consolidação em fraturas diafisárias. Métodos Este estudo de coorte observacional prospectivo incluiu pacientes adultos com fraturas diafisárias dos principais ossos longos previamente submetidas a fixação interna sem consolidação. Após o tratamento definitivo, os pacientes foram avaliados periodicamente por seis meses, com realização seriada de ultrassonografia, determinação da concentração de FA e radiografias para verificar a presença de consolidação. Resultados Após um aumento inicial em sete semanas, os níveis de FA voltaram ao valor normal em pacientes com fraturas consolidadas, mas continuaram elevados nos casos de ausência de consolidação. Da mesma forma, após uma elevação do índice de resistência (IR) vascular em cerca de 12 semanas em todos os pacientes, o IR diminuiu nos casos que progrediram para consolidação, mas continuou alto até as 24 semanas em fraturas não consolidadas. Os casos com ausência de consolidação ainda apresentavam calo hipoecogênico às 24 semanas, que não se converteu no calo hiperecogênico observado nos casos que progrediram para consolidação. Conclusão Alterações significativas sugestivas de consolidação foram simultaneamente observadas nas radiografias, na ultrassonografia e na concentração de FA durante o período de acompanhamento. No entanto, a realização seriada de exames da concentração de FA e de ultrassonografia durante o acompanhamento indicou o progresso da consolidação da fratura. Seu papel no monitoramento da ausência de consolidação é mais complementar do que suplementar à radiografia.

Distracción osteogénica del tercio medio facial. Reporte de dos casos clínicos / Distraction osteogenesis of the midface. Two clinical cases report

La corrección quirúrgica de grandes asimetrías faciales son un desafío para el equipo quirúrgico. En dichos casos, los resultados de la corrección utilizando técnicas quirúrgicas convencionales son limitados, lo que hace necesario la utilización de otras herramientas terapéuticas. La distracción ósea (DO) es una de ellas, pues permite el estiramiento controlado del callo óseo previamente creado mediante osteotomías. La DO permite realizar cambios en el posicionamiento óseo de gran envergadura, favoreciendo el crecimiento óseo como mecanismo de acción. El objetivo de este artículo es describir los resultados quirúrgicos de DO intraoral del tercio medio facial utilizado en dos pacientes con severas asimetrías faciales asociadas a malformaciones de origen genético. Se relata el diagnóstico, la planificación, las herramientas tecnológicas utilizadas, técnica quirúrgica y los resultados obtenidos.

Surgical correction of major facial asymmetries is a challenge for the surgical team. In such cases, treatment results from conventional surgical techniques are limited, which requires using other therapeutic tools. Bone distraction is one of them, as it allows controlled stretching of the bone callus previously developed through osteotomies. Distraction osteogenesis allows making changes in large bone positioning, favoring bone growth as an action mechanism. This article aims to describe the surgical results of intraoral distraction osteogenesis of the midface used in two patients with severe facial asymmetries related to congenital malformations. Diagnosis, planning, technological tools, surgical techniques, and results obtained are explained.