Microalgae possess high photosynthetic efficiency, robust adaptability, and substantial biomass, serving as excellent biological resources for large-scale cultivation. Malic enzyme (ME), a ubiquitous metabolic enzyme in living organisms, catalyzes the decarboxylation of malate to produce pyruvate, CO₂, and NAD(P)H, playing a role in multiple metabolic pathways including energy metabolism, photosynthesis, respiration, and biosynthesis. In this study, we identified the Scenedesmus quadricauda malic enzyme gene (SqME) and its biological functions, aiming to provide excellent target genes for the genetic improvement of higher plants. Based on the RNA-seq data from S. quadricauda under the biofilm cultivation mode with high CO₂ and light energy transfer efficiency and small water use, a highly expressed gene (SqME) functionally annotated as ME was cloned. The physicochemical properties of the SqME-encoded protein were systematically analyzed by bioinformatics tools. The subcellular localization of SqME was determined via transient transformation in Nicotiana benthamiana leaves. The biological functions of SqME were identified via genetic transformation in Nicotiana tabacum, and the potential of SqME in the genetic improvement of higher plants was evaluated. The ORF of SqME was 1 770 bp, encoding 590 amino acid residues, and the encoded protein was located in chloroplasts. SqME was a NADP-ME, with the typical structural characteristics of ME. The ME activity in the transgenic N. tabacum plant was 1.8 folds of that in the wild-type control. Heterologous expression of SqME increased the content of chlorophyll a, chlorophyll b, and total chlorophyll by 20.9%, 26.9%, and 25.2%, respectively, compared with the control. The transgenic tobacco leaves showed an increase of 54.0% in the fluorescence parameter NPQ and a decrease of 30.1% in Fo compared with the control. Moreover, the biomass, total lipids, and soluble sugars in the transgenic tobacco leaves enhanced by 20.5%, 25.7%, and 9.5%, respectively. On the contrary, the starch and protein content in the transgenic tobacco leaves decreased by 22.4% and 12.2%, respectively. Collectively, the SqME-encoded protein exhibited a strong enzymatic activity. Heterologous expressing of SqME could significantly enhance photosynthetic protection, photosynthesis, and biomass accumulation in the host. Additionally, SqME can facilitate carbon metabolism remodeling in the host, driving more carbon flux towards lipid synthesis. Therefore, SqME can be applied in the genetic improvement of higher plants for enhancing photosynthetic carbon fixation and lipid accumulation. These findings provide scientific references for mining of functional genes from S. quadricauda and application of these genes in the genetic engineering of higher plants.
Nicotiana/genetics* ; Photosynthesis/physiology* ; Malate Dehydrogenase/biosynthesis* ; Plant Leaves/genetics* ; Scenedesmus/enzymology* ; Carbon Cycle/genetics* ; Lipid Metabolism/genetics* ; Plants, Genetically Modified/metabolism*

Treatment of calcaneal fractures has always been the focus of many clinical researchers.The goals of traditional surgical treatment are not only to restore the integrity of calcaneal articular surface but also to reconstruct the anatomy of the calcaneus.More importantly,we need to reduce postoperative soft tissue swelling and incidence of postoperative complications.In recent years,scholars have reported satisfactory clinical efficacy and prognosis resulting from a sinus tarsal approach for treatment of calcaneal fractures.This paper reviews the latest research progress concerning the sinus tarsal approach for treatment of calcaneal fractures at home and abroad,intending to provide helpful information for the clinical surgeons.

Objective: To analyze the genome characteristics of an avian influenza A (H9N2) virus isolated from an 11-month-old infant, and to look for possible sources of infection. Methods: Throat swabs were collected from an infant with influenza-like illness in influenza sentinel surveillance hospitals and isolated for influenza viruses using cells. The isolates were identified for influenza virus types and subtypes by the method of hemagglutination assay, hemagglutination inhibition assay and fluorescence PCR. Whole genome sequencing of the isolated virus was carried out. The genome nucleic acid sequences and the deduced amino acid sequences were analyzed by comparing the phylogenetic trees which were constructed by bioinformatics software. Results: A seasonal un-typed influenza virus was isolated from the infant with influenza like illness. With fluorescent PCR method , it was identified as H9N2 subtype of avian influenza virus and the case was confirmed as a human infected with an avian influenza A(H9N2) virus. Epidemiological studies revealed that the case had no clear history of poultry contact and exposure. Blast analysis shows that eight segments of the viral genome are avian origin, and 97.5%-99.8% homology with that of viruses isolated from the live-poultry markets. The virus belongs to G57 genotype, deduced amino acid sequence analysis shows that the virus has typical low pathogenic avian influenza characteristics. Conclusions Although the case does not have a clear history of contact or exposure to poultry, molecular traceability suggests that possible sources of infection may be still from poultry.

Objective To study changes of serum inflammatory indicators,plasma and urine free amino acid of patients with chronic obstructive pulmonary disease.Methods A total of 62 patients with chronic obstructive pulmonary disease in our hospital from June 2014 to April 2016 were selected as observation group,62 healthy with physical examination in our hospital during the same time were selected as control group,the serum inflammatory indicators,plasma and urine free amino acid of two groups were compared,and the above indicators of observation group with different GOLD classifications and stages were compared too.Results The serum inflammatory indicators of observation group were all higher,and plasma free amino acid indicators were all lower than that of control group,and the seruminflammatory indicators and plasma free amino acid indicators of observation group with different GOLD classifications and stages showed significant differences (P ＜ 0.05),while the urine free amino acid indicators of two groups had no significant differences (P ＞ 0.05).Conclusion The change of serum inflammatory indicators and plasma free amino acid of patients with chronic obstructive pulmonary disease are obvious,and the influence of severity degree and disease stages for the expression are greater,while the urine free amino acid have no obvious fluctuation,so the detection value of serum inflammatory indicators and plasma free amino acid of patients with chronic obstructive pulmonary disease are higher.

Objective To study changes of serum inflammatory indicators,plasma and urine free amino acid of patients with chronic obstructive pulmonary disease.Methods A total of 62 patients with chronic obstructive pulmonary disease in our hospital from June 2014 to April 2016 were selected as observation group,62 healthy with physical examination in our hospital during the same time were selected as control group,the serum inflammatory indicators,plasma and urine free amino acid of two groups were compared,and the above indicators of observation group with different GOLD classifications and stages were compared too.Results The serum inflammatory indicators of observation group were all higher,and plasma free amino acid indicators were all lower than that of control group,and the seruminflammatory indicators and plasma free amino acid indicators of observation group with different GOLD classifications and stages showed significant differences (P ＜ 0.05),while the urine free amino acid indicators of two groups had no significant differences (P ＞ 0.05).Conclusion The change of serum inflammatory indicators and plasma free amino acid of patients with chronic obstructive pulmonary disease are obvious,and the influence of severity degree and disease stages for the expression are greater,while the urine free amino acid have no obvious fluctuation,so the detection value of serum inflammatory indicators and plasma free amino acid of patients with chronic obstructive pulmonary disease are higher.

Objective To study the effects of anti -cancer drug NSC319726 on the gene expression of ovarian cancer.Methods The data that NSC319726 acted on the ovarian cancer cell lines TOV112D was down-loaded from the GEO database and then the bioinformatical analysis was conducted .Results NSC319726 in-creased 246 genes including MMP3,CXCR4 et al and decreased 198 genes such as PBX1 in ovarian cancer cell lines.GO analysis indicated that six GO items were related to metabolism .Pathway analysis revealed that NSC319726 could affect the circadian clock and MMP signaling pathway .Conclusion NSC319726 could affect the expression of multiple functional genes in ovarian cancer cell lines and might be a potential drug targeting to the precursor-B cell acute lymphoblastic leukemia .Meanwhile , it should be noted that NSC 319726 may have effects on the tumor metastasis and human sleep .Therefore,further research needs to be conducted before the clinical use .

The purpose of this paper is to investigate that the effect of cyclic biaxial mechanical strain on proliferation and synthetic function in the osteoblasts isolated from 3 month-old normal female Sprague-Dawley (SD) rats and osteoporotic rats. The osteoblasts were cultured in F-12 medium contained with 10% fetal bovine serum (FBC) and grown to subconfluency in Flexercell apparatus in a humidified incubator with 5% CO2 and 95% air at 37 degrees C. Mechanical strain was applied to the cells for periods of 30 min, 2, 4 and 8 hours every day,lasting 2 days. The amplitude of mechanical strain applied to the cells were 400, 1000 and 4000 microm strain respectively, at a frequency of one hertz (1 H). Unstrained cells were used as control. The results showed that proliferation activity of osteoblast in osteoporotic rats are higher than that in normal rats without mechanical strain the mechanical strain can elevate the proliferation activity and the synthetic function of osteoblast from normal rats at 400, 1000 microm strain. However, the mechanical strain increased significantly the proliferation in the osteoblasts and suppressed obviously the synthetic function in the osteoblasts at 4000 microm strain. The mechanical strain don't affect the proliferation activity and the synthetic function of osteoblast from osteoporotic rats at 400 microm strain. The mechanical strain decrease the proliferation activity of osteoblast from osteoporotic rats at 1000 microm strain. The mechanical strain can elevate the proliferation activity and the synthetic function of osteoblast from osteoporotic rats at 4000 microm strain. In our study, the reaction of the osteoblasts from normal rats and osteoporotic rats to the mechanical stimulation suggested that there are more highly sensitive to the mechanical stimulation in the osteoblasts from normal rats than that from osteoporotic rats.
Animals ; Apoptosis ; Cell Division ; Cell Separation ; Cells, Cultured ; Female ; Osteoblasts ; metabolism ; pathology ; Osteocalcin ; biosynthesis ; Osteoporosis ; pathology ; Rats ; Rats, Sprague-Dawley ; Stress, Mechanical

The purpose of this paper is to investigate the probable molecular mechanism in mechano-transduction of the regulation of integrins and the effects of cyclic biaxial mechanical strain on proliferation and synthetic function in the osteoblasts isolated from 3-month-old female Sprague-Dawley (SD) rats. The osteoblasts were cultured in F-12 medium contained with 10% fetal bovine serum(FBS) and grown to subconfluency in Flexercell type I dishes in a humidified incubator with 5% CO2 and 95% air at 37 degrees C. Mechanical strains were applied to the cells for periods of 30 min, 2, 4 and 8 hours every day, lasting 2 days. The amplitude of mechanical strain applied to the cells were 400, 1,000 and 4,000 mu strain respectively, at a frequency of one hertz(1 Hz). Unstrained cells were used as control. The expression of integrins alpha 2, beta 1, beta 3 on the membrane of osteoblasts and proliferation activity of osteoblasts were studied with Flow Cytometry(FCM). The content of osteocalcin, carboxyterminal propeptide of type-I procollage(PICP), total protein secreted by osteoblastes were detected with the isotope labelling method. The results showed that there are actual expressions of integrins alpha 2, beta 1, beta 3 on the membrane of osteoblasts without mechanical strain and that the expression of integrins beta 1 is highest. The mechanical strain increased the expression of integrins alpha 2, beta 1, beta 3 on the membrane of osteoblasts, but the strain-related up-regulation of expression of integrins alpha 2, beta 1, beta 3 are different in various amplitude and different duration of mechanical stains. The up-regulation of expression of integrins beta 3 is most sensitive to mechanical strain. The up-regulation of expression of integrins alpha 2, beta 1, beta 3 is higher at 4,000 mu strain than at 400, 1,000 mu strain. The mechanical strain can elevate the proliferation activity and the synthetic function of osteoblast at 400, 1,000 mu strain. However, the mechanical strain increased significant the proliferation in the osteoblasts and suppressed obviously the synthetic function in the osteoblasts. In the present study, the reaction of the osteoblasts in 3 month-old rat to the mechanical stimulation suggested that 1) expressions of integrins alpha 2, beta 1, beta 3 were increased in a amplitude of strain-dependent manner; 2) the changes of expression of integrins alpha 2, beta 1, beta 3 relate close to the changes of the proliferation and synthetic function of the osteoblasts. Low amplitude of strain can increase the proliferation and the synthetic function of the osteoblasts along with up-regulation of expression of integrins alpha 2, beta 1, beta 3; while higher amplitude of strain elevated significantly the proliferation of osteoblasts and suppressed obviously the synthetic function of the osteoblasts along with up-regulation of expression of integrins alpha 2, beta 1, beta 3. The amplitude of 4,000 mu strain is an optimal amplitude as stimulus for up-regulation of expression of integrins alpha 2, beta 1, beta 3 on the membrane of osteoblasts and increase the proliferation activity, but decrease the synthetic function of osteoblasts in the present study. Accordingly it indicates that integrins have a important role in regulation of signal transduction pathway in osteoblasts as a result of mechanical strain.
Animals ; Cell Division ; Cells, Cultured ; Female ; Integrin alpha2 ; biosynthesis ; Integrin beta1 ; biosynthesis ; Integrin beta3 ; biosynthesis ; Mechanics ; Osteoblasts ; metabolism ; physiology ; Rats ; Rats, Sprague-Dawley