1.Evidence-based guideline for clinical diagnosis and treatment of acute combination fractures of the atlas and axis in adults (version 2023)
Yukun DU ; Dageng HUANG ; Wei TIAN ; Dingjun HAO ; Yongming XI ; Baorong HE ; Bohua CHEN ; Tongwei CHU ; Jian DONG ; Jun DONG ; Haoyu FENG ; Shunwu FAN ; Shiqing FENG ; Yanzheng GAO ; Zhong GUAN ; Yong HAI ; Lijun HE ; Yuan HE ; Dianming JIANG ; Jianyuan JIANG ; Weiqing KONG ; Bin LIN ; Bin LIU ; Baoge LIU ; Chunde LI ; Fang LI ; Feng LI ; Guohua LYU ; Li LI ; Qi LIAO ; Weishi LI ; Xiaoguang LIU ; Yong LIU ; Zhongjun LIU ; Shibao LU ; Fei LUO ; Jianyi LI ; Yong QIU ; Limin RONG ; Yong SHEN ; Huiyong SHEN ; Jun SHU ; Yueming SONG ; Tiansheng SUN ; Jiang SHAO ; Jiwei TIAN ; Yan WANG ; Zhe WANG ; Zheng WANG ; Xiangyang WANG ; Hong XIA ; Jinglong YAN ; Liang YAN ; Wen YUAN ; Jie ZHAO ; Jianguo ZHANG ; Yue ZHU ; Xuhui ZHOU ; Mingwei ZHAO
Chinese Journal of Trauma 2023;39(4):299-308
The acute combination fractures of the atlas and axis in adults have a higher rate of neurological injury and early death compared with atlas or axial fractures alone. Currently, the diagnosis and treatment choices of acute combination fractures of the atlas and axis in adults are controversial because of the lack of standards for implementation. Non-operative treatments have a high incidence of bone nonunion and complications, while surgeries may easily lead to the injury of the vertebral artery, spinal cord and nerve root. At present, there are no evidence-based Chinese guidelines for the diagnosis and treatment of acute combination fractures of the atlas and axis in adults. To provide orthopedic surgeons with the most up-to-date and effective information in treating acute combination fractures of the atlas and axis in adults, the Spinal Trauma Group of Orthopedic Branch of Chinese Medical Doctor Association organized experts in the field of spinal trauma to develop the Evidence-based guideline for clinical diagnosis and treatment of acute combination fractures of the atlas and axis in adults ( version 2023) by referring to the "Management of acute combination fractures of the atlas and axis in adults" published by American Association of Neurological Surgeons (AANS)/Congress of Neurological Surgeons (CNS) in 2013 and the relevant Chinese and English literatures. Ten recommendations were made concerning the radiological diagnosis, stability judgment, treatment rules, treatment options and complications based on medical evidence, aiming to provide a reference for the diagnosis and treatment of acute combination fractures of the atlas and axis in adults.
2.Effect of Gualou Xiebai Banxiatang on Mitochondrial Dysfunction and AMPK/PGC-1α Signaling Pathway in Rats with Ischemic Myocardial Injury
Yingying TAN ; Hui WANG ; Yanbing WANG ; Hong LI ; Chengcheng MA ; Qi ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(1):9-17
ObjectiveTo observe the effect of Gualou Xiebai Banxiatang on mitochondrial dysfunction and adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK)/peroxlsome proliferator-activated receptor-γ coactlvator-1α (PGC-1α) signaling pathway in rats with ischemic myocardial injury. MethodSeventy male SD rats were used in this experiment. Six rats from them were randomly selected as the control (CON) group, and the others were given high fat diet combined with isoproterenol injection (5 mg·kg-1·d-1, 7 d) to induce the rat model of ischemic heart disease based on hyperlipidemia. Successfully modeled rats were then randomly divided into model (MOD) group, high-dose Gualou Xiebai Banxiatang (GXBD-H) group, medium-dose Gualou Xiebai Banxiatang (GXBD-M) group, low-dose Gualou Xiebai Banxiatang (GXBD-L) group, and metoprolol (MET) group. Rats in the GXBD-H, GXBD-M, and GXBD-L groups were given 11.2, 5.6, 2.8 g·kg-1·d-1 Gualou Xiebai Banxiatang, those in the MET group were given 2.6 mg·kg-1·d-1 metoprolol, and those in the CON and MOD groups were given equal volume of pure water for 28 d. Hemodynamics were measured in rats by cardiac catheterization. Transmission electron microscopy was used to analyze myocardial mitochondria. Serum brain natriuretic peptide (BNP) and cardiac troponin T (cTnT) levels were detected by enzyme-linked immunosorbent assay (ELISA). Mitochondrial membrane potential assay kit (JC-1 method) was applied for detecting mitochondrial membrane potential. The changes in the mitochondrial DNA copy number were measured by real-time quantitative polymerase chain reaction (Real-time PCR). The content of adenosine triphosphate (ATP) in myocardial tissues was determined by spectrophotometer. The expression levels of p-AMPK, AMPK, PGC-1α, nuclear respiratory factor 1 (NRF1), and mitochondrial transcription factor A (TFAM) in myocardium was detected by Western blot. ResultAs compared with the CON group, left ventricular end-systolic pressure (LVESP) and left ventricular end-diastole pressure (LVEDP) in the MOD group were significantly increased (P<0.05, P<0.01), and +dp/dtmax and -dp/dtmax were significantly decreased (P<0.01). In the MOD group, cardiac index and myocardial interstitial fibrosis area were significantly increased (P<0.01), accompanied by mitochondrial damage, serum BNP, cTnT, and malondialdehyde (MDA) were significantly increased (P<0.01), and serum superoxide dismutase (SOD) level was significantly decreased (P<0.01). The myocardial mitochondrial membrane potential, DNA copy number, and ATP level were significantly decreased (P<0.01), and the protein expression levels of p-AMPK/AMPK, PGC-1α, NRF1, and TFAM in myocardial tissues were significantly decreased in the MOD group (P<0.01). Compared with the MOD group, the GXBD-H and GXBD-M groups significantly improved LVESP, LVEDP, +dp/dtmax, and -dp/dtmax (P<0.05, P<0.01), significantly decreased heart index and myocardial interstitial fibrosis area (P<0.05, P<0.01), and alleviated mitochondrial damage. In the GXBD-H and GXBD-M groups, serum BNP, cTnT, and MDA were decreased significantly (P<0.05, P<0.01), serum SOD level was increased significantly (P<0.05), and myocardial mitochondrial membrane potential, DNA copy number, and ATP level were significantly increased (P<0.05, P<0.01). The protein levels of p-AMPK/AMPK, PGC-1α, NRF1, and TFAM in myocardial tissues were significantly increased in the GXBD-H and GXBD-M groups (P<0.05, P<0.01). ConclusionGualou Xiebai Banxiatang has the effects of reducing the changes in cardiac function and myocardial pathology of rats with myocardial injury, inhibiting mitochondrial dysfunction, and up-regulating the protein expression levels of p-AMPK/AMPK, PGC-1α, NRF1, and TFAM in myocardial tissues. This study provides new laboratory evidence for in-depth exploration of the mechanism of this classical compound in preventing and treating myocardial injury.
3.Expert consensus on implementation strategy of awake prone positioning for non-intubated patients in China (2023).
Yuanyuan MI ; Zheyi CAI ; Jing LIU ; Fei TIAN ; Liping YANG ; Lei BAO ; Shanbing HOU ; Su GU ; Li LI ; Xueli ZHOU ; Yun XU ; Shumei ZHANG ; Xiaoxia FU ; Xiaodi LI ; Chuansheng LI ; Liang SUN ; Xiaohong ZHANG ; Hong QI ; Shiying YUAN ; Liqun ZHU ; Haiyan HUANG ; You SHANG
Chinese Critical Care Medicine 2023;35(4):337-351
The awake prone position plays an important role in the treatment of hypoxemia and the improvement of respiratory distress symptoms in non-intubated patients. It is widely used in clinical practice because of its simple operation, safety, and economy. To enable clinical medical staff to scientifically and normatively implement prone position for awake patients without intubation, the committees of consensus formulation, guided by evidence-based methodology and Delphi method, conducted literature search, literature quality evaluation and evidence synthesis around seven topics, including indications and contraindications, evaluation, implementation, monitoring and safety management, termination time, complication prevention and health education of awake prone position. After two rounds of expert letter consultation, Expert consensus on implementation strategy of awake prone positioning for non-intubated patients in China (2023) was formulated, and provide guidance for clinical medical staff.
Humans
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Consensus
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Prone Position
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Wakefulness
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China
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Dyspnea
4.Regulating mechanism of dynein heavy chain on autophagic degradation of α-synuclein in Parkinson disease model cells
Jing LU ; Bo SUN ; Shikun CAI ; Xiangyang TIAN ; Quan CHEN ; Bin QI ; Lei WANG ; Xiangyuan CHEN ; Zhenjie SUN
Chinese Journal of Behavioral Medicine and Brain Science 2023;32(10):871-878
Objective:To explore the regulatory mechanism of α-synuclein in the degradation of autophagy-lysosome pathway(ALP) in Parkinson disease(PD) model cells after interference or overexpression of dynein heavy chain(Dynhc) gene.Methods:SH-SY5Y cells were divided into control group, PD group, Dynhc interference group, Dynhc overexpression group, and Dynhc interference+ rapamycin group according to experimental requirements.Using Western blot to detect Dynhc, α-synuclein, microtubule-associated protein l light chain 3 (LC3), lysosome-associated membrane protein 2 (LAMP2), tubulin, dynein activator protein p150, and kinesin KIF5B.Flow cytometry was used to detect the level of cell apoptosis.Immunoconfocal microscopy was used to observe the structure of tubulin and the co-localization of LC3 and LAMP.SPSS 23.0 software was used for statistical analysis.One-way ANOVA was used for inter group comparisons, and further pairwise comparisons were conducted by LSD- t test. Results:There were statistically significant differences in the expression of α-synuclein, autophagy-related proteins, microtubules, and microtubule-related proteins among cells in the 5 groups(all P<0.001). The protein expression levels of Dynhc, α-synuclein, LC3, LAMP2, p150, and KIF5B in the PD group were higher than those in the control group (all P<0.05). The protein levels of Dynhc, LAMP2, tubulin and p150 in the Dynhc interference group were lower than those in the PD group (all P<0.05), while the protein levels of α-synuclein, LC3 and KIF5B were higher than those in the PD group (all P<0.05). The protein levels of α-synuclein, LC3, and KIF5B in the Dynhc overexpression group were lower than those in the PD group (all P<0.05), while the protein levels of Dynhc, LAMP2 and p150 were higher than those in the PD group (all P<0.05). The protein level of LC3 in the Dynhc interference+ rapamycin group was higher than that in the Dynhc interference group ( P<0.05). There were no statistically significant differences in the protein levels of Dynhc, α-synuclein, LAMP2, microtubule protein, p150 and KIF5B compared to the Dynhc interference group (all P>0.05). Compared with the control group, the cell apoptosis rate in PD group increased((12.77±1.66)%, (7.64±1.45)%), the microtubule morphology remained unchanged, and autophagosomes fused more with lysosomes. Compared with the PD group, the cell apoptosis rate of Dynhc overexpression group decreased, and there was no significant change in microtubule structure, and there was more fusion between autophagosomes and lysosomes.Compared with the PD group, the cell apoptosis rat of Dynhc interference group increased((18.45±1.91)%), and the microtubule morphology was sparse, and there was less fusion between autophagosomes and lysosomes. Compared with the PD group, the Dynhc overexpression group showed a decrease in cell apoptosis rate ((9.95±1.56)%), no significant changes in microtubule structure, and more fusion between autophagosomes and lysosomes.Compared with the Dynhc interference group, the Dynhc interference+ rapamycin group showed no significant changes in cell apoptosis rate ((19.05±2.46)%), microtubule morphology, and fusion of autophagosomes and lysosomes. Conclusion:Dynhc can reduce cell apoptosis by enhancing cell ALP function, increasing the degradation of α-synuclein and maintaining of microtubule structure integrity.
5.Effects of SKP1 gene interference on apoptosis and regulation mechanism of ubiquitin-proteasome system in Parkinson's disease cell model
Zhenjie SUN ; Qiang TONG ; Qi GAO ; Ting LU ; Quan CHEN ; Xiangyang TIAN ; Bo SUN
Chinese Journal of Behavioral Medicine and Brain Science 2022;31(11):976-983
Objective:To investigate the effect of interfering S-phase kinase associated protein 1 (SKP1) gene on apoptosis in Parkinson's disease(PD) cell model induced by 1-methyl-4-phenyl-pyridine ion (MPP+ ) and the mechanism of ubiquitin proteasome system degradation of α-synuclein (α-syn) influence.Methods:SH-SY5Y cells were divided into control group, MPP+ group, SKP1 interference group, and SKP1 interference+ MG132(UPS inhibitor) group.The cells in the control group were cultured normally. The cells in the latter three groups were incubated with MPP+ (0.5 mmol/L) for 24 h as PD model cells.The cells in SKP1 interference group were transfected with lentivirus SKP1-siRNA, and the cells in SKP1 interference+ MG132 group were transfected with lentivirus SKP1 siRNA and added with MG132 (0.5 μmol/L) for 24 h. The protein levels and mRNA levels of SKP1, microtubule-associated protein light chain 3 (LC3), lysosome-associated membrane protein (LAMP), α-syn, ubiquitin activating enzyme E1 (UBE1), parkin, and p27 in cells were detected by Western blot and RT-PCR.Flow cytometry was used to detect cell apoptosis and cycle level, and CCK-8 method was used to detect cell proliferation level.Co-immunoprecipitation method was used to explore the interaction between SKP1 and p27. SPSS 23.0 software was used for statistical analysis. One-way ANOVA was used for comparison among groups, and LSD test was used for further pairwise comparison.Results:RT-PCR and Western blot results showed that the mRNA levels and protein levels of autophagy related proteins and ubiquitin related proteins LC3, LAMP2, α-syn, UBE1, parkin and p27 in the four groups were statistically significant(mRNA: F=99.155, 43.028, 138.464, 28.200, 22.009, 28.147, all P<0.05; F=245.517, 157.634, 315.920, 2 336.472, 477.429, 2 350.201, all P<0.05). The mRNA and protein levels of LC3, Lamp2, α-syn and p27 in SKP1 interference group were lower than those in MPP+ group (all P<0.05), while the mRNA and protein levels of UBE1 and parkin were higher than those in MPP+ group (all P<0.05). The mRNA and protein levels of LC3, α-syn and p27 in SKP1 interference+ MG132 group were higher than those in SKP1 interference group (all P<0.05), and the mRNA and protein levels of UBE1 and parkin were lower than those in SKP1 interference group (all P<0.05). The results of flow cytometry and CCK-8 method showed that the apoptosis rate and cell inhibition rate among the four groups were significantly different( F=2 749.420, 171.508, both P<0.05). The apoptosis rate of SKP1 interference group was lower than that of MPP+ group ((8.22±0.25)%, (15.30±0.21)%, P<0.05), while the cell inhibition rate of SKP1 interference group was lower than that of MPP+ group((26.31±3.73)%, (55.05±3.84)%, P<0.05). The apoptosis rate of SKP1 interference+ MG132 group ((9.49±0.07)%) was higher than that of SKP1 interference group, and the cell inhibition rate ((36.06±2.85)%) was higher than that of SKP1 interference group (both P<0.05). The results of immunoprecipitation method showed that P27 decreased after SKP1 immunoprecipitation. Conclusion:After SKP1 gene was interfered, the autophagy function of PD cells decreased, which may be related to parkin promoting α-syn ubiquitination, activating UBE1/ Parkin-mediated UPS pathway to degrade α-syn, and mediating P27 to inhibit apoptosis.
6.Characteristics of bicuspid aortic valve aortopathy and association with valvular phenotype and dysfunction
Jing SUN ; Cheng SUN ; Zhihui HOU ; Hongxia QI ; Xiangyang QIAN ; Zhe ZHENG
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2022;29(06):729-735
Objective To summarize the characteristics of bicuspid aortic valve (BAV) aortopathy and analyze the association between aortopathy and BAV phenotype and patterns of valvular dysfunction. Methods Clinical data of 191 patients who underwent the first aortic valve replacement in Fuwai Hospital from June 2017 to March 2018 were retrospectively analyzed, including 143 males and 48 females with an average age of 53.91±12.52 years. All patients underwent multidetector computed tomography (MDCT) and echocardiography before the operation, excluding patients with aortic coarctation. The BAV aortopathy phenotype was classified during operation. The characteristics of BAV aortopathy were analyzed by cluster and artificial analysis. BAV anatomic phenotype was divided into two types according to the direction of valve opening: BAV-AP and BAV-LR. Results Four distinct BAV aortopathy phenotypes were identified: a common type (n=70, 36.6%), with no dilation or mild dilation of aorta; a root type (n=24, 12.6%), with predominant dilatation of aortic sinus; an ascending aorta type (n=72, 37.7%), with predominant dilatation of ascending aorta; an arch type (n=25, 13.1%), with predominant dilatation of aortic arch dilatation. The root type was mainly in young patients, while the arch type was mainly in elderly patients (P<0.05). BAV-AP and aortic insufficiency were most prevalent in root type, while BAV-LR and aortic stenosis were most prevalent in arch type (P<0.05). There were 111 (58.1%) patients undergoing aortic surgery, and the coincidence rate of BAV aortopathy phenotype and aortic surgery was 80.6%. Conclusion According to the location of aortic dilation, BAV aortopathy can be divided into four types. There is an association between BAV aortopathy and valvular phenotype and dysfunction.
7.Identification of Decoction Pieces and Standard Decoction of Ophiopogonis Radix and Liriopes Radix by TLC
Yu HAO ; Qi-shu JIAO ; Yan-yan ZHOU ; Ru-na JIN ; Chun-miao XUE ; Shou-gang SHI ; Zheng-jun HUANG ; Yun-tao DAI
Chinese Journal of Experimental Traditional Medical Formulae 2020;26(15):124-129
Objective:There were 92 kinds of compound preparations containing Ophiopogonis Radix in the 2015 edition of
8.Quality evaluation methods for standard decoction of Nelumbinis Folium.
Xue-Yuan WANG ; Yun-Tao DAI ; Ru-Na JIN ; Qi-Shu JIAO ; Shou-Gang SHI ; Zheng-Jun HUANG ; Shuo-Sheng ZHANG
China Journal of Chinese Materia Medica 2020;45(6):1363-1367
The quality evaluation method for standard decoction of Chinese herbal slices is the basis for the quality evaluation of granules and preparations of classical formula(decoction)of traditional Chinese medicine. This study aimed to establish a method for the determination of quercetin-3-O-glucuronic acid in Nelumbinis Folium(NF)and its standard decoction, so as to provide reference for the quality control of NF and its standard decoction. Fifteen batches of representative NF were collected to prepare standard decoction, and the parameters of dry extract rate, transfer rate of index component, and pH value were calculated. HPLC was used to establish the content determination method for quercetin-3-O-glucuronic acid in NF and its standard decoction. The concentration range of quercetin-3-O-glucuronic acid in the standard decoction of NF was 1.09-3.06 g·L~(-1), while the concentration range of nuciferine was 0.01-0.17 g·L~(-1). The average extraction rate of NF standard decoction was(14.4±2.6)%, the average transfer rate of quercetin-3-O-glucuronic acid was(70.7±18.6)%, and the average transfer rate of nuciferine was(9.6±5.4)%. Compared with Nuciferine, quercetin-3-O-glucuronic acid had a high content and stable transfer rate in standard decoction, and was recommended to be the quality control marker for NF and its standard decoction. This paper establishes a quality evaluation method for NF standard decoction, and can provide reference for the quality control of all preparations derived from NF and its decoction.
Chromatography, High Pressure Liquid
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Drugs, Chinese Herbal/chemistry*
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Flowers/chemistry*
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Medicine, Chinese Traditional
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Nelumbo/chemistry*
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Quality Control
9.Antibiotics-mediated intestinal microbiome perturbation aggravates tacrolimus-induced glucose disorders in mice.
Yuqiu HAN ; Xiangyang JIANG ; Qi LING ; Li WU ; Pin WU ; Ruiqi TANG ; Xiaowei XU ; Meifang YANG ; Lijiang ZHANG ; Weiwei ZHU ; Baohong WANG ; Lanjuan LI
Frontiers of Medicine 2019;13(4):471-481
Both immunosuppressants and antibiotics (ABX) are indispensable for transplant patients. However, the former increases the risk of new-onset diabetes, whereas the latter impacts intestinal microbiota (IM). It is still unclear whether and how the interaction between immunosuppressants and ABX alters the IM and thus leads to glucose metabolism disorders. This study examined the alterations of glucose and lipid metabolism and IM in mice exposed to tacrolimus (TAC) with or without ABX. We found that ABX further aggravated TAC-induced glucose tolerance and increased insulin secretion. Combined treatment resulted in exacerbated lipid accumulation in the liver. TAC-altered microbial community was further amplified by ABX administration, as characterized by reductions in phylum Firmicutes, family Lachnospiraceae, and genus Coprococcus. Analyses based on the metagenomic profiles revealed that ABX augmented the effect of TAC on microbial metabolic function mostly related to lipid metabolism. The altered components of gut microbiome and predicted microbial functional profiles showed significant correlation with hepatic lipid accumulation and glucose disorders. In conclusion, ABX aggravated the effect of TAC on the microbiome and its metabolic capacities, which might contribute to hepatic lipid accumulation and glucose disorders. These findings suggest that the ABX-altered microbiome can amplify the diabetogenic effect of TAC and could be a novel therapeutic target for patients.
10.Histomorphometryof the vertebra and its correlations with MRI signal intensity in rabbits with fluoride treatment
Haifeng XU ; Jin QI ; Jinshen WANG ; Xiangyang XU ; Qi ZHOU ; Zhenguo ZHAO ; Yong LU ; Kemin CHEN ; Yongli ZHANG ; Lianfu DENG
Chinese Journal of Orthopaedics 2018;38(19):1195-1203
Objective To analyze histomorphometrical characteristics of the bone and bone marrow tissues of the lumbar vertebrae in rabbits with fluoride treatment,and its correlation with signal intensity of MRI.Methods Forty New Zealand albino rabbits aged three months old were randomly divided into fluoride exposure of 30 cases and control of 10 cases,male and female,half each.One hundred milligrams of sodium fluoride were added to the municipal water each liter (fluoride content 100 mg/L) as drinking waterto fluorine for 180 days.Twenty-four of 30 cases with fluoride exposure had complete data (male10 casesand female14 cases).The same municipal water was used as control drinking water (fluoride content < 0.9 mg/L).Eight of 10 cases with control had complete data (male andfemale in half).Twenty-four cases with fluoride treatment and complete data were classified into sensitive and resistant type according to the MRI signal intensity of the lumbar vertebra.Histomorphometrics of the vertebra and its correlation with the MRI signal intensity,and sensitivity in early diagnosis of osteofluorosis and feasibility of susceptibility to osteofluorosis detected with MRI were analyzed.Results Theratios of trabecular bone volume (BV),hematopoietic cell volume (HV) and fluid volume (FV) in bone marrow tissue to total cavernous tissue volume (TT) in group with fluoride treatment were 18.3%±2.6%,45.2%±6.0% and 10.4%±5.7% respectively.These were 14.5%±2.8%,36.3%±7.3% and 6.2%±2.1% in control group respectively.These parameters in fluoride group were significantly increased compared to control group.The ratio 26.0%± 8.0% of adipocyte volume (AV) to TV in fluoride group was significantly lower than that 43.3%±5.6% in control group.Two of 24 cases with fluoride exposure (8.3%,2/24) were sensitive and the remaining 22 (91.7%,22/24) were in resistance.The valuesof BV/TT,HV/TV and FV/TV were considered to be sensitive,resistant and control from large to small,while AV/TV value were opposite.A comparison resuhs of signal intensity in MRI showed that vertebra T1WI contrast to noise ratio (CNR) in the sensitive was the minimum (3.0±0.8),followed by resistance (21.3±3.8) andmaximum in the control (28.3±3.1),but CNR of FsT2WIwas opposite.There were positive associations between T1WI and AV/TV,FV/TV and BV/TV,and between FsT2WI and FV/TV and BV/ TV.There were inverse associationsbetween FsT2WI and AV/TV.Theoptimal threshold value of the vertebra T1WI CNR was 23.2 or lessin early diagnosis of skeletal fluorosis,with sensitivity of 83.3% and specificity of 100%.FsT2WI was 5.7 or more,with sensitivity of 45.8% and specificity of 100%.Conclusion The pathogenesis of osteofluorosis is relative to changes in bone marrow microenvironment and cells number in bone marrow tissue,and is correlated to MRI signal intensity.

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