1.The responsive characteristics of phytochrome genes to photoperiod, abiotic stresses and identification of their key natural variation sites in foxtail millet (Setaria italica L.).
Xiaoping JIA ; Bo ZHANG ; Zhanxiang HE ; Jianfeng LI ; Xiaomei ZHANG ; Di GE ; Zhenshan WANG ; Luman SANG ; Zhiwei SONG
Chinese Journal of Biotechnology 2022;38(5):1929-1945
The responsive patterns of phytochrome gene family members to photoperiod and abiotic stresses were comparatively analyzed and the favorable natural variation sites of these genes were identified. This would help understand the mechanism of phytochrome gene family in photoperiod-regulated growth and development and abiotic stress response. In addition, it may facilitate the molecular marker assisted selection of key traits in foxtail millet. In this study, we used RT-PCR to clone three phytochrome genes SiPHYA, SiPHYB and SiPHYC from ultra-late maturity millet landrace variety 'Maosu'. After primary bioinformatics analysis, we studied the photoperiod control mode and the characteristics of these genes in responding to five abiotic stresses including polyethylene glycol (PEG)-simulated drought, natural drought, abscisic acid (ABA), high temperature and NaCl by fluorescence quantitative PCR. Finally, we detected the mutation sites of the three genes among 160 foxtail millet materials and performed haplotype analysis to determine the genes' functional effect. We found that the cloned cDNA sequences of gene SiPHYA, SiPHYB and SiPHYC were 3 981, 3 953 and 3 764 bp respectively, which contained complete coding regions. Gene SiPHYB and SiPHYC showed closer evolutionary relationship. Photoperiod regulated all of the three genes, but showed more profound effects on diurnal expression pattern of SiPHYB, SiPHYC than that of SiPHYA. Under short-day, when near heading, the expression levels of SiPHYA and SiPHYB were significantly lower than that under long-day, indicating their roles in suppressing heading of foxtail millet under long-day. SiPHYB and SiPHYC were responsive to PEG-simulated drought, natural drought, ABA and high temperature stresses together. SiPHYA and SiPHYB responded differently to salt stress, whereas SiPHYC did not respond to salt stress. Re-sequencing of 160 foxtail millet materials revealed that SiPHYB was highly conservative. Two missense mutations of SiPHYA, such as single nucleotide polymorphism (SNP) 7 034 522C→T and SNP7 036 657G→C, led to delaying heading and increasing plant height. One missense mutation of SiPHYC, such as SNP5 414 823G→T, led to shortening heading under short-day and delaying heading under long-day, as well as increasing plant height and panicle length regardless of photo-thermal conditions. Photoperiod showed different regulatory effects on SiPHYA, SiPHYB and SiPHYC. SiPHYB and SiPHYC jointly responded to various abiotic stresses except for the salt stress. Compared with the reference genotype, mutation genotypes of SiPHYA and SiPHYC delayed heading and increased plant height and panicle length.
Gene Expression Regulation, Plant
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Photoperiod
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Phytochrome/metabolism*
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Plant Proteins/metabolism*
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Setaria Plant/metabolism*
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Stress, Physiological/genetics*
2.Recent advance in paradoxical herniation
Jiawei HE ; Qi YANG ; Liang ZHANG ; Zhanxiang WANG
Chinese Journal of Neuromedicine 2022;21(3):291-293
Paradoxical herniation (PH) is a rare complication after decompressive craniectomy, which manifests as that intracranial contents collapse due to atmospheric pressure higher than intracranial pressure and gravity after decompression, and the brain parenchyma eventually herniates through the tentorial notch or the foramen magnum. Contrary to the traditional treatment of cerebral hernia, the intracranial pressure in PH patients should be increased by early cranioplasty or other measures. This article reviews the clinical manifestations, mechanism, diagnoses and treatments of PH, and provides further reference for clinical work.
3.Effect of p38 mitogen-activated protein kinase pathway on hydrocephalus and aquaporin 4 expression in rats
Zhangyu LI ; Xin GAO ; Jianyao MAO ; Sifang CHEN ; Guowei TAN ; Zhanxiang WANG
Chinese Journal of Neuromedicine 2022;21(4):333-340
Objective:To observe the regulatory effect of p38 mitogen-activated protein kinase (p38 MAPK) on aquaporin 4 (AQP4) in rats after hydrocephalus, and to explore its significance in hydrocephalus prevention.Methods:Fifty SD rats were randomly divided into sham-operated group ( n=10), hydrocephalus group ( n=20), and hydrocephalus+inhibitor (SB203580) group (SB group, n=20). The rat models of hydrocephalus in the latter two groups were prepared by intracerebroventricular injection of kaolin suspension; rats in the sham-operated group were injected with same amount of normal saline into the lateral ventricle. The p38 MAPK specific inhibitor SB203580 (10 mg/kg) was intraperitoneally injected into the rats of SB group on the 8 th d of modeling for 7 consecutive d; same volume of dimethylsulfoxide was given to the rats of hydrocephalus group on the 8 th d of modeling for 7 consecutive d; rats in the sham-operated group did not give any treatment. The severity of hydrocephalus in these rats was observed by MRI. The inflammatory factor tumor necrosis factor (TNF)-α level in the cerebrospinal fluid was detected by enzyme-linked immunosorbent assay (ELISA). The AQP4 and TNF-α mRNA expressions were detected by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). The phosphorylated p38 MAPK and AQP4 expressions in the periventricular brain tissues were detected by Western blotting and immunohistochemistry. Results:No hydrocephalus developed in sham-operated group and hydrocephalus developed in the latter two groups. As compared with sham-operated group, hydrocephalus group and SB group had significantly increased lateral ventricle volume, significantly aggravated periventricular edema, significantly higher EVAN's index, and statistically increased brain water content ( P<0.05). Two weeks after modeling, the TNF-α expression levels in cerebrospinal fluid of sham-operated group, hydrocephalus group and SB group were (20.49±0.96), (42.04±3.17), and (28.00±3.71) pg/mL, respectively, with significant differences ( F=186.000, P<0.001); the TNF-α expression level in SB group was significantly higher than that in sham-operated group and significantly lower than that in hydrocephalus group ( P<0.05). Two weeks after modeling, the TNF-α and AQP4 mRNA expression levels in brain tissues of the three groups were significantly different ( P<0.05); the TNF-α and AQP4 mRNA expression levels in hydrocephalus group were significantly higher than those in sham-operated group and SB group ( P<0.05). Correlation analysis showed that there was a positive linear correlation between AQP4 mRNA expression and TNF-α mRNA expression in hydrocephalus group ( r=0.511, P=0.026), and there was a positive linear correlation between AQP4 protein expression and phosphorylated p38 MAPK protein expression in hydrocephalus group and SB group ( r=0.560, P=0.013; r=0.463, P=0.030). Immunohistochemical staining results showed that AQP4 expression was abundant in glial cells of the three groups; the p38 MAPK distribution was uniform and non-polar; the phosphorylated p38 MAPK protein expression in the hydrocephalus group was significantly higher than that in the sham-operated group, and that in the SB group returned to the level of the sham-operated group. Conclusion:The p38 MAPK pathway is involved in the positive regulation of AQP4 expression, which could be inhibited by SB203580.
4.Clinical application analysis of a method for locating scalp projection of intracranial lesions based on neuroimaging
Zhongjie SHI ; Liwei ZHOU ; Shuo WANG ; Chaofan FAN ; Guowei TAN ; Zhanxiang WANG
Chinese Journal of Surgery 2022;60(6):606-610
Objective:To explore the feasibility of a method based on neuroimaging and surface markers for locating scalp projection of intracranial lesions.Methods:The clinical data of 46 patients who were used ‘double-circle method’ for locating scalp projection of intracranial lesions at Department of Neurosurgery,the First Affiliated Hospital of Xiamen University from January to June 2021 were retrospective analyzed. All patients with 2 electrodes(artificial fiducials) randomly attached to scalp had been examed thin-layer brain CT. The distances from the center of each fiducial to the root of the nose and tragus were measured through the images. A compass was used to draw two arcs with the root of nose and the tragus as the center and the pre-measured distance as the radius on patient′s scalp. Then two arcs′ intersection on the scalp was the fiducial. The method was named ‘double-circle method’. Two neurosurgeons were arranged to perform fiducial identification with double-circle method, and record the error between the result and the actual fiducial point.Independent sample t test was used for data comparison, and Kappa test was used to analysis the inter-group consistency. Results:Ninety-two fiducial points of 46 patients were collected. Time consuming of doctor A was (8.1±2.3) minutes(range:5 to 15 minutes)and doctor B was (8.9±3.5) minutes(range:4 to 17 minutes).The positioning error from the doctor A was (4.4±2.4)mm(range:0 to 12 mm) and doctor B was(4.2±2.6) mm(range:0 to 14 mm)( t=-0.575, P=0.567),the difference was not statistically significant. The Kappa value of the consistency test of error between two doctors was 0.517( P=0.001).The consistency was moderate.Eight patients used ‘double-circle method’ and neuronavigation for locating scalp projection of intracranial lesions at the same time. The diameter of the lesions was (3.8±0.9)cm (range: 2.6 to 5.1 cm), and the positioning error of the ‘double-circle method’ and navigation was (4.0±1.9) mm(range: 1 to 6 mm), and all patients were confirmed to be accurately located during surgery. Conclusion:‘Double-circle method’ is a simple,convenient and accurate way in locating intracranial lesions and has certain clinical significance.
5.Clinical application analysis of a method for locating scalp projection of intracranial lesions based on neuroimaging
Zhongjie SHI ; Liwei ZHOU ; Shuo WANG ; Chaofan FAN ; Guowei TAN ; Zhanxiang WANG
Chinese Journal of Surgery 2022;60(6):606-610
Objective:To explore the feasibility of a method based on neuroimaging and surface markers for locating scalp projection of intracranial lesions.Methods:The clinical data of 46 patients who were used ‘double-circle method’ for locating scalp projection of intracranial lesions at Department of Neurosurgery,the First Affiliated Hospital of Xiamen University from January to June 2021 were retrospective analyzed. All patients with 2 electrodes(artificial fiducials) randomly attached to scalp had been examed thin-layer brain CT. The distances from the center of each fiducial to the root of the nose and tragus were measured through the images. A compass was used to draw two arcs with the root of nose and the tragus as the center and the pre-measured distance as the radius on patient′s scalp. Then two arcs′ intersection on the scalp was the fiducial. The method was named ‘double-circle method’. Two neurosurgeons were arranged to perform fiducial identification with double-circle method, and record the error between the result and the actual fiducial point.Independent sample t test was used for data comparison, and Kappa test was used to analysis the inter-group consistency. Results:Ninety-two fiducial points of 46 patients were collected. Time consuming of doctor A was (8.1±2.3) minutes(range:5 to 15 minutes)and doctor B was (8.9±3.5) minutes(range:4 to 17 minutes).The positioning error from the doctor A was (4.4±2.4)mm(range:0 to 12 mm) and doctor B was(4.2±2.6) mm(range:0 to 14 mm)( t=-0.575, P=0.567),the difference was not statistically significant. The Kappa value of the consistency test of error between two doctors was 0.517( P=0.001).The consistency was moderate.Eight patients used ‘double-circle method’ and neuronavigation for locating scalp projection of intracranial lesions at the same time. The diameter of the lesions was (3.8±0.9)cm (range: 2.6 to 5.1 cm), and the positioning error of the ‘double-circle method’ and navigation was (4.0±1.9) mm(range: 1 to 6 mm), and all patients were confirmed to be accurately located during surgery. Conclusion:‘Double-circle method’ is a simple,convenient and accurate way in locating intracranial lesions and has certain clinical significance.
6.Role of 3D printing positioning guide in neurosurgery
Zhongjie SHI ; Xin GAO ; Liwei ZHOU ; Shuo WANG ; Jianfeng GUO ; Guowei TAN ; Zhanxiang WANG
Chinese Journal of Neuromedicine 2021;20(10):1039-1042
Objective:To explore the application value of individualized three-dimensional (3D) printing positioning guides in localization and resection of intracranial lesions.Methods:Fifteen patients with intracranial space occupying lesions underwent resection in our hospital from March 2021 to May 2021 were selected in our study. Brain images by CT and MRI as raw data were used to design individual positioning guides. The positioning guides were placed on the patient's skin before resection to mark the location and boundary of the lesions with a marker, and neuro-navigation was used to verify the accuracy. During the resection, the location of the lesions was identified through microscope by the surgeons. Postoperative CT and MRI were used to evaluate the lesion resection.Results:The individualized positioning guides of 15 patients fit the skin well, and the skin incision and bone window were designed to meet the surgical requirements. All surgeries were completed in one time, and the lesion tissues were successfully removed. During the surgeries, the skin incision was not adjusted for secondary expansion. Brain MRI reexamination within 48 h of surgery showed that the lesions of 11 patients with tumors were removed satisfactorily (total resection in 9 and subtotal resection in 2); brain CT reexamination within 12 h showed that the clearance rate of hematomas in 3 patients was above 80% and that in 1 patient was 70%. No patients had cerebrospinal fluid leakage, intracranial hematoma, intracranial infection or other serious complications. All patients recovered well during the 1-3 months of outpatient/telephone follow-up.Conclusion:The positioning method with personalized 3D printing guides is simple and convenient, enjoying accurate positioning results, which can assist the clinicians to optimize the preoperative planning, optimize the surgical incision design, and is worthy of promotion and application in primary hospitals.
7.Preliminary study of serum 2019-nCoV IgM and IgG antibodies in the diagnosis of COVID-19
Ping LI ; Zhiyong LI ; Silin ZHAO ; Qiong LI ; Yan HU ; Yufeng CHEN ; Fan YI ; Qichen XIE ; Zhaoqiong ZENG ; Changjuan DENG ; Zhanxiang WANG ; Xiaobing XIE
Chinese Journal of Laboratory Medicine 2020;43(4):352-357
Objective:To analyze the clinical value of serum 2019 novel coronavirus (2019-nCoV) immunoglobulin M (IgM) and immunoglobulin G (IgG) antibodies in the diagnosis of COVID-19.Methods:A total of 116 patients diagnosed with NCP in the First Affiliated Hospital of Hunan University of Chinese Medicine and the First Affiliated Hospital of Xiamen University were enrolled from January to February 2020 as the disease group. A total of 134 cases, including 84 non-NCP inpatients and 50 healthy individuals served as the control group. Serum samples from all subjects were collected. A fully-automated chemiluminescence immunoassay analyzer was used to detect the concentration of 2019-nCoV IgM and IgG antibodies in serum. The sensitivity and specificity of the 2019-nCoV IgM and IgG antibody single test and combined detection were compared using the χ 2 test. χ 2 test and Wilcoxon′s rank sum test were used to compare the positive rates and concentrations of IgM and IgG antibodies in NCP patients before and after their 2019-nCoV nucleic acid tests turning negative, respectively. The change trend of 2019-nCoV antibody concentration in the process of NCP patients was analyzed by Wilcoxon′s rank sum test. Results:The sensitivity of 2019-nCoV IgG (90.5%, 105/116) was higher than that of 2019-nCoV IgM (75.9%, 88/116), the difference was statistically significant (χ 2=8.91, P<0.05); The specificity of 2019-nCoV IgG (99.3%,133/134) was higher than that of 2019-nCoV IgM (94.0%, 126/134), the difference was statistically significant (χ 2=5.63, P<0.05). The sensitivity (89.7%,87/97) of 2019-nCoV IgM combined with IgG was higher than that of 2019-nCoV IgM, the difference was statistically significant (χ 2=6.89, P<0.05). The specificity (100%, 125/125) of 2019-nCoV IgM combined with IgG was higher than that of 2019-nCoV IgM, the difference was statistically significant (χ 2=7.70, P<0.05). After 2019-nCoV nucleic acid test converted to negative, the positive rate (9/17) and concentration [13.0 (4.9, 24.7) AU/ml] of serum 2019-nCoV IgM antibody were significantly lower than those when the nucleic acid test was positive, positive rate (15/17) and concentration [29.5 (14.0, 61.3) AU/ml], respectively (χ 2=5.10, Z=-3.195, both P<0.05). In the course of NCP, patients′ serum samples were collected from the first day of diagnosis to every three days, three times in total. The first 2019-nCoV IgM and IgG antibody concentrations [19.4 (12.4, 63.7) AU/ml, 105.8 (74.8, 126.1) AU/ml, respectively] were significantly higher than the second concentrations [15.8 (7.1, 40.3)AU/ml, 80.5 (66.7, 105.9) AU/ml], Z were-2.897,-3.179, both P<0.05. Conclusions:2019-nCoV IgG antibody has a good application value in the diagnosis of NCP. The concentration of 2019-nCoV IgM antibody has a certain correlation with the detection of 2019-nCoV nucleic acid. The combination of 2019-nCoV IgM and IgG antibodies with 2019-nCoV nucleic acid test may be the best laboratory index for the diagnosis of NCP at present.
8.Genetic analysis of a pedigree affected with tuberous sclerosis complex caused by a novel mutation of TSC1.
Shide ZHANG ; Zhanxiang WANG ; Rong LIN ; Jingjun ZHAO
Chinese Journal of Medical Genetics 2019;36(10):961-964
OBJECTIVE:
To analyze the clinical phenotype of a Chinese pedigree affected with Tuberous sclerosis complex (TSC) and explore pathogenic mutations of TSC1 and TSC2 gene.
METHODS:
Unique clinical phenotypes,the results of imaging, examination of the proband and special family history, collectively, made the constellation of features of TSC. Genomic DNA was obtained from six affected and eight unaffected members of the family and potential mutations of the TSC1 and TSC2 genes were detected by PCR-amplification of the exons and exon-intron boundaries and direct sequencing. A total of 150 normal unrelated individuals were used as controls.
RESULTS:
Genetic analysis documented the presence of a heterozygous mutation, c.1781_1782delTG (p.Val594GlyfsX11), in the exon 15 of TSC1 gene within all the patients of the family. This mutation was not observed in the eight unaffected family members or in the 150 unrelated control subjects from the same population , or the Human Gene Mutation Database (HGMD) and had completely co-segregated with the disease phenotype in the family.
CONCLUSION
The c.1781_1782delTG mutation of TSC1 gene may be responsible for the tuberous sclerosis complex in this family. The data presented in the present study are of significance to clinicians, as well as genetic counselors, and may provide new clues for molecular diagnosis of this disease..
DNA Mutational Analysis
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Humans
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Mutation
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Pedigree
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Tuberous Sclerosis
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genetics
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Tuberous Sclerosis Complex 1 Protein
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genetics
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Tuberous Sclerosis Complex 2 Protein
9. Genetic analysis of a pedigree affected with tuberous sclerosis complex caused by a novel mutation of TSC1
Shide ZHANG ; Zhanxiang WANG ; Rong LIN ; Jingjun ZHAO
Chinese Journal of Medical Genetics 2019;36(10):961-964
Objective:
To analyze the clinical phenotype of a Chinese pedigree affected with Tuberous sclerosis complex(TSC) and explore pathogenic mutations of
10.Awake craniotomy of gliomas in eloquent areas: an analysis of 19 cases
Xi CHEN ; Liang ZHANG ; Jinli SUN ; Weichao JIANG ; Sifang CHEN ; Guowei TAN ; Zhanxiang WANG ; Hongming BAI
Chinese Journal of Neuromedicine 2017;16(12):1270-1274
Objective To discuss the techniques and methods of surgery for brain gliomas located in eloquent areas at awake anesthesia. Methods Nineteen patients with brain gliomas in eloquent areas, admitted to our hospital from December 2014 to May 2017, were operated under awake anesthesia with neuronavigation and intraoperative ultrasonography for locating the lesions and intraoperative direct electrical stimulation for functional mapping of the eloquent areas. All patients were followed up from 3 to 18 months; the surgical efficacies were analyzed. Results Of 19 patients, 18 (94.74%) were achieved awake and alert during brain mapping and resection of the tumors;17 (89.47%) were detected the motor areas by intraoperative direct electrical stimulation, 6 (31.58%) were detected the sensory cortex and 12 (63.16%) were detected language related cortex. Of 19 patients, MR imaging 2-3 months after surgery indicated that 5 (26.32%) received total resection of lesions, 9 (47.37%) subtotal resection of lesions and 5 (26.32%) partial resection of lesions. Seven patients (36.84%) had transitory postoperative aphasia, 4 (21.05%) were with transitory postoperative dyskinesia and one (5.26%) with permanent dyskinesia. Conclusion Comprehensive applications of awake anesthesia, neuronavigation, intraoperative ultrasonography and intraoperative direct electrical stimulation technologies allow maximum safe resection of gliomas in eloquent areas and protection of brain function.

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