1.Bibliometric and visual analysis of domestic and foreign scoliosis orthoses
Yanli YUAN ; Yuejun PAN ; Tianmin GUAN ; Kai CHENG ; Xiangheng WANG
Chinese Journal of Tissue Engineering Research 2024;33(33):5396-5402
BACKGROUND:Scoliosis mainly refers to sequence abnormalities in the coronal,sagittal,and axial positions of the spine,with a Cobb angle of≥10°.The patients may experience symptoms such as unequal shoulder height and back asymmetry.Severe cases may affect the patient's cardiopulmonary function,thereby affecting their daily life.Conservative treatment can control the progression of scoliosis and avoid later surgery.Scoliosis orthosis is currently a commonly used and effective treatment measure in conservative treatment. OBJECTIVE:To summarize and analyze the current research status,hotspots,and trends of scoliosis orthoses both domestically and internationally,providing reference for related research. METHODS:Using bibliometrics and visual analysis as tools,and using a comparison between China and foreign countries as a method,this paper analyzes the literature on scoliosis orthosis journals in the past decade.Based on bibliometrics,the current status of research on scoliosis orthoses is determined.Citespace software is used to analyze key words and identify the current hotspots and future trends in scoliosis orthosis research. RESULTS AND CONCLUSION:(1)At present,the number of literature on scoliosis orthoses is still on a fluctuating upward trend.China and the United States are the main countries for research,with a literature share of over 40%.However,the average citation rate of foreign language literature by Chinese scholars is relatively low.(2)The basic fields of domestic research are mainly surgery and pediatrics,while orthotics and clinical neurology are mainly studied abroad.Among them,there is also a certain number of documents in domestic Chinese medicine,indicating that China is also engaged in the combination of Chinese and Western treatment of scoliosis.The National Natural Science Foundation of China has the highest proportion in the aspect of Chinese and foreign literature,reflecting the importance of the fund attaches to the research of scoliosis orthosis.(3)The authors with the highest number of publications are Qiu Yong and Negrini Stefano,and the most published institutions are the Spinal Surgery Department of Gulou Hospital affiliated to Nanjing University Medical College and UDICE-French Research University.Domestic and foreign authors and institutions have certain communications about this,but not closely,which requires relevant institutions and scholars to further explore and study.(4)From the research hotspots and future trends,the main treatment type is adolescent idiopathic scoliosis,while the production method of the short-column side bending orthosis is three-dimensinoal printing,and the main treatment index is convex progression.The ultimate purpose of treatment is to improve the quality of life of the patients.
2.Correlation between lesion volume ratio and cognitive function in ischemic leukoaraiosis
Na SUN ; Jianfeng WANG ; Tianmin GUAN ; Aiqi WANG ; Xuemei WANG ; Lizhen ZHONG ; Xueying CHENG ; Hua ZHAO
Chinese Journal of Postgraduates of Medicine 2022;45(1):31-36
Objective:To investigate the relationship between the volume ratio of ischemic leukoaraiosis (LA) and cognitive level and arterial perfusion.Methods:Fifty-four patients, who was hospitalized in Dalian Central Hospital and diagnosed as LA clinically during the time of March to December in 2012, were selected to collect the information of the volume ratio of white matter disease, MoCa score and the average flow rate of carotid artery. The correlation between the volume ratio of white matter disease and MoCa score, cognitive impairment and the average flow rate of carotid artery were analyzed.Results:The volume ratio of LA lesions was negatively correlated with MOCA score ( r = -0.59, P<0.01); the volume ratio of LA lesions was negatively correlated with the mean flow rate of internal carotid artery ( r = -0.37, P<0.01). Quantity order of the area under receiver operating characteristic (ROC) curve of MoCA cognitive subgroup was as following: delayed memory (1.000)> visual space/executive function (0.970) = abstract force (0.970)> language ability (0.960)> attention (0.888). Conclusions:The larger the volume ratio of leukopathy in LA patients, the more serious the cognitive impairment, especially the cognitive impairment of impairment of memory delay, visual space/executive function, abstract ability and language ability.
3.The application of respiratory exerciser tri-ball in pulmonary rehabilitation patients with chronic obstructive pulmonary disease
Tianmin GAO ; Quanchang ZHOU ; Shicong HUANG ; Cheng SHU ; Li LUO ; Jiahua CHEN ; Xiaorong YU ; Chenxu LI ; Min DAI
Chongqing Medicine 2015;(32):4514-4516
Objective To discuss the function of respiratory exerciser tri‐ball in pulmonary rehabilitation (PR) patients with chronic obstructive pulmonary disease(COPD) .Methods Prospectie case‐control study was used in COPD patients ,the patients were randomly divided into three groups ,60 COPD patients (group A) using respiratory exerciser TRI‐BALL ,58 COPD patients (group B) using traditional pursed lips ventral breathing training ,and 58 COPD patients(group C) using general internal medicine treatment .Results Group A :compared with before breath training ,the increases of FEV1/FEV1 predicted (% ) and MVV/MVV predicted (% ) and the decrease of quality of life score (QOL) were statistically significant after breath training (P<0 .01) ,but not for FEV1/FVC(% )(P>0 .05) .Group B :compared with before breath training ,the decrease of QOL was statistically significant (P<0 .01) ,but not for FEV1/FEV1 predicted (% ) ,MVV/MVV predicted (% ) and FEV1/FVC (% )(P>0 .05) .Compared with control group after breath training ,the increases of FEV1/FEV1 predicted (% ) and MVV/MVV predicted (% ) and the decrease of quality of life score (QOL) were statistically significant in group A (P<0 .01) .Compared with control group after breath training , the decrease of quality of life score (QOL) were statistically significant in group B (P<0 .01) ,but not for FEV1/FEV1 predicted (% ) and MVV/MVV predicted (% ) (P>0 .05) .Compared the changes of pulmonary function test(PFT) index and QOL between group A and B ,the increments of FEV1/FEV1 predicted (% ) and MVV/MVV predicted (% ) were statistically significant in group A(P<0 .01) ,but not for QOL(P>0 .05) .Conclusion It is useful to improve the pulmonary function and quality of life in patients with COPD using respiratory exerciser tri‐ball .It is more effective than traditional pursed lips ventral breathing training ,due to the equipment is very small ,cheap ,easy to quantify training and convenient for household use ,it is worth to be popularized in primary hospital .
4.Enhancement of distribution of dermal multipotent stem cells to bone marrow in rats of total body irradiation by platelet-derived growth factor-AA treatment
Zhaowen ZONG ; Yongchuan REN ; Yue SHEN ; Yonghua CHEN ; Xinze RAN ; Chunmeng SHI ; Tianmin CHENG
Chinese Journal of Radiological Medicine and Protection 2011;31(4):433-436
Objective To observe whether dermal multipotent stem cells (dMSCs) treated with platelet-derived growth factor-AA ( PDGF-AA )could distribute more frequently to the bone marrow in rats of total body irradiation (TBI).Methods Male dMSCs were isolated and 10 μg/L PDGF-AA was added to the culture medium and further cultured for 2 h.Then the expression of tenascin-C were examined by Western blot, and the migration ability of dMSCs was assessed in transwell chamber.The pre-treated dMSCs were transplanted by tail vein injection into female rats administered with total body irradiation, and 2 weeks after transplantation, real-time PCR was employed to measure the amount of dMSCs in bone marrow.Non-treated dMSCs served as control.Results PDGF-AA treatment increased the expression of tenascin-C in dMSCs, made (1.79 ± 0.13) × 105 cells migrate to the lower chamber under the effect of bone marrow extract, and distributed to bone marrow in TBI rats, significantly more than ( 1.24 ± 0.09) ×105 in non-treated dMSCs (t = 8.833, P < 0.0l ).Conclusions PDGF-AA treatment could enhance the migration ability of dMSCs and increase the amount of dMSCs in bone marrow of TBI rats after transplantation.
5.CXCR4 gene transfection enhancing the distribution of dMSCs to the wounded skin of rats with combined wound and irradiation injury
Zhaowen ZONG ; Tianmin CHENG ; Xinze RAN ; Yongping SU ; Shiwu DONG ; Nan LI ; Junping WANG ; Guoping AI ; Zheng LI
Chinese Journal of Radiological Medicine and Protection 2009;29(4):351-354
Objective To observe whether the transplanted dermal multipotent stem cells(dMSCs)transfected by adenovirus vector of CXCR4(Adv-CXCR4)can distribute more frequently to the wound of rats with combined wound and irradiation injury.Methods dMSCs transfected by Adv-CXCR4(group A),or transfected by adenovirus vector of green fluorescent protein(group B),and non-transfected dMSCs were labeled with 3H-TdR and then transplanted into combine-injured rats.The amount of dMSCs in wound were determined by liquid scintillation,and wounds healing process was observed by measuring the remaining wound area.Results From the 5th day after transplantation,the amount of dMSCs in the wound of group A accounted for 1.95%-3.85% of the total transplanted dMSCs,significantly greater than those in group B and group C,which accounted for 1.07%-1.86% of the total transplanted dMSCs.The remaining wound area in group A was smaller than those in group B and group C from day 12 after injury,and the healing time of group A was 1.5 day ahead than group B and group C.Conclusions dMSCs transfected by Adv-CXCR4 distributes more frequently to the wound of combine-injured rats and could accelerate wound healing.
6.Effect of long-term depleted uranium ingestion on testosterone production and the expression of StAR and P450sec mRNAs in rats
Rong LI ; Yanbing LENG ; Jiong REN ; Jing LIU ; Guoping AI ; Hui XU ; Yongping SU ; Tianmin CHENG
Chinese Journal of Radiological Medicine and Protection 2009;29(1):13-16
Objective To explore the effect of long-term depleted uranium (DU)ingestion on testosterone production in rats, and its involvement mechanism. Methods Male and female rats (F0 and F1 respectively) for 160 days, respectively. The contents of testosterone (T), luteinizing hormone (LH), and follicle stimulating hormone (FSH) in serum were detected in 20 months of F0 generations, and 15 months of F1 generations. RT-PCR was used to analyze the levels of StAR mRNA and P450scc mRNA. Results Compared with the normal control group, the testosterone contents in exposed F0 and F1 generations increased, the lowest was 51.73 U/L, but those of LH and FSH decreased. The expression of StAR mRNA in the low-doze group of F1 generation (StAR/β-actin = 1.35) was up-regulated, down-regulated for other groups.compared with the normal control group (P450scc/β-actin = 0. 313), the expression of P450scc mRNA in the low- and high-dose groups of F0 generation were decreased (P450scc/β-actin = 0.21), and those in the low- and high-dose groups of F1generation were increased (P450scc/β-actin = 0.623) (P ≤ 0.01). Conclusion Long-term DU exposure inhibit the male reproduction by intervening the sexual hormone production through down-regulated the expression of StAR mRNA and P450scc roRNA.
7.Soluble expression and purification of human alpha-defensin-5 in Escherichia coli.
Aiping WANG ; Yongping SU ; Tianmin CHENG ; Zhongmin ZOU ; Junping WANG
Chinese Journal of Biotechnology 2008;24(2):291-296
DNA fragment containing human alpha-defensin 5 mature peptide (mHD-5) coding sequence with biased codons of E. coli was amplified by PCR, which was subsequently cloned into the plasmid pMAL-p2x in order to create pMAL-p2x-mHD-5 expression vector. The plasmid pMAL-p2x-mHD-5 was transferred into engineered strain BL21(DE3) to express heterogeneous fusion protein (MBP-mHD-5). The soluble MBP-mHD-5 targeted protein inducible expressed by IPTG was accounted for about 30% under optimized conditions. The recombinant mHD-5 (rmHD-5) peptide was successfully purified through a separation process including affinity chromatography, Factor Xa digestion and ion exchange chromatography. The bioactivity of rmHD-5 was examined by bacteria-inhibition tests in liquid culture. The growth of E. coli ATCC25922 was dramatically suppressed with an inhibition rate of 90%, with the presence of 62.5 microg/mL rmHD-5 in the media. These results indicate that the strategy of soluble expression of fusion protein in E. coli can be a useful and practical way to produce bioactive defensins.
Cloning, Molecular
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Escherichia coli
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genetics
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metabolism
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Humans
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Recombinant Fusion Proteins
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biosynthesis
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genetics
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isolation & purification
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Solubility
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Transformation, Bacterial
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alpha-Defensins
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biosynthesis
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genetics
8.Morphological study on the growth of human amniotic membrane loaded with porcine bone marrow-derived mesenchymal stem cells
Guohe YAN ; Guoping AI ; Daijie WANG ; Zhongmin ZOU ; Xinze RAN ; Junping WANG ; Rong LI ; Yongping SU ; Tianmin CHENG
Chinese Journal of Tissue Engineering Research 2007;11(15):2985-2989
BACKGROUND: Human amniotic membrane (HAM) contains various ingredents such as collagen, glycoprotein,proteoglycan, integrin and laminated body, and so on, and expresses many kinds of growth factors and mRNA-associated proteins. And these ingredents can supply abundant nutriments for cellular proliferation and differentiation, and benefit cells to grow and propagate. Whether or not HAM can load porcine bone marrow-derived mesenchymal stem cells (BMSCs) to well grow on it deserves to be further investigated.OBJECTIVE: To set up a method of tissue engineering of human amniotic membrane loading porcine BMSCs and observe the morphological characteristics of growth and proliferation of BMSCs seeded on HAM.DESIGN: Randomized controlled observation.SETTING: State Key Laboratory of Trauma, Burn and Combined Injury, General Institute of Combined Injuries, Academy of Preventive Medicine, Third Military Medical University of Chinese PLA.MATERIALS: This experiment was carried out in the State Key Laboratory of Trauma, Burn and Combined Injury,General Institute of Combined Injuries, Academy of Preventive Medicine, Third Military Medical University of Chinese PLA between January and November 2003. Three Guizhou minipigs of either gender, aged 2 to 3 months, weighing from 6 to 8 kg, were provided by the Experimental Animal Center, Third Military Medical University of Chinese PLA. Main reagent:ISCOVE'S modified DULBECCO'S medium (IMDM) culture medium (Hyclone, USA); high-quality fetal bovine serum PAA (Germany); haematoxylin (China); Eosin B (Sigma, USA) and OCT embedding medium (USA). Main instruments: BX51 stereoscopic fluorescence microscope (Olympus, JaPan); IX70 inverted fluorescence microscope (Olympus, Japan);cryostat (2700-Frigcut, Germany); myeloid puncture needle (Jiangsu); superclean bench (Sujing Bloc Antai Company);CO2 constant-temperature incubator (QUEUE, USA).METHODS: HAM was prepared as previously described. The BMSCs of Guizhou minipigs isolated and cultured according to method described previously were primarily cultured and passaged, then they were inoculated to the stromal surface of HAM at different densities (0.84×105 cells/cm2,1.54×105 cells/cm2,2.75×105 cells/cm2); The growth and proliferation of BMSCs of different densities were observed under an inverted microscope and scanning electron microscope; BMSCs of the second or the third passages were inoculated on HAM held with tissue-holding device at a density of 1.54×105 cells/cm2, and they were cultured for 18 days at most. The HAM was daily rolled, sliced and stained by HE for observing the growth of BMSCs loaded on HAM under the light, scanning and transmission electron microscopes.MAIN OUTCOME MEASURES: The growth of BMSCs on HAM was examined at different densities and different time points.RESULTS: ① Comparison of growths of BMSCs promoted by different densities of HAM: BMSCs,which were planted on HAM at the density of 0.84×105 cells/cm2 were irregular and scattered under an invert microscope. Distances between BMSCs were biggish. BMSCs seeded on HAM at the density of 1.54×105 cells/cm2 were regular in arrangement and moderate in density, with clear cell outline and good cell activity before 24 hours, and seeded at the density of 2.75×105 cells/cm2 were congested with many nonattached cells and the longer the growing time of the cells was, the more the cellular debris were observed. BMSCs,which were planted on HAM at the density of 0.84×105 cells/cm2 under the scanning electron microscope, scatted on HAM presented in shapes of irregular, long, thin and flat polygon. Their membrane protuberances presented in shapes of thick and thin, and the distances between cells were biggish. BMSCs,which were planted on HAM at the density of 1.54×105 cells/cm2 have similar appearance of their bodies and membrane protuberances, and the membrane protuberances were more compared with the BMSCs planted at the density of 0.84×105 cells/cm2. Their membrane protuberances intercrossed each other, and the margin of some BMSCs overlapped each other. BMSCs planted at the density of 2.75×105 cells/cm2, arraved on HAM crowdedly and overlappedly with many debris. Their membrane protuberances were not obviously. The margin of some BMSCs was overlapped.② Comparisonof growths of BMSCs promoted by HAM at different time points: Under the inverted microscope, the BMSCs adhered quickly to HAM after being incubated for about 30 minutes. All of BMSCs adhered to HAM within 24 hours, and formed monolayer on it within 48 hours, and grew densely on HAM after being cultured for 4 to18 days. Under the light and electron microscopes, HE results revealed that BMSCs adhered tightly and grew on HAM in different arrays, such as emitting, whirlpool or parallel,and their nuclei located in middle, dense in staining, were big and clear. The shapes of BMSCs were comparatively consistent on HAM. HAM loaded with BMSCs grew 4 days, and BMSCs covered HAM completely. The densities of BMSCs on HAM were suitable, and their bodies were large, and presented irregular, long,thin and flat polygon under the scanning electron microscope. The margin of some BMSCs overlapped each other. The protuberances of cellular membrane of BMSCs were abundant in the shapes of thick and thin. Some protuberances intercrossed each other in the shape of net. BMSCs adhered tightly to HAM through these protuberances. HAM loading BMSCs grow 4 days; most of BMSCs grew on HAM in double layers with the shapes of cambiform under the transmission electron microscope, Their nucleoli were clear. The protuberances of cellular membrane of BMSCs, which situated at two sides of nuclei and overlapped each other, were long. Most of chromatins of BMSCs were autosome.Abundant organell such as rough endoplasmic reticulum (RER),mitochondria could be observed in BMSCs.CONCLUSION:HAM is able to promote the proliferation of BMSCs significantly. BMSCs may be cultured on HAM ex vivo.HAM is a good carrier of BMSCs.
9.Effect of cervical sympathetic ganglia block on the mortality of mice with combined radiation and burn injury and its possible mechanism
Jianhua LU ; Chong SHI ; Yongping SHU ; Tianmin CHENG ; Zhiyong DU
Chinese Journal of Tissue Engineering Research 2006;10(34):177-180
BACKGROUND: Cervical sympathetic ganglia block accelerates the re covery of the homeostasis of organic nervous-endocrine-immune system, butit is still unclear whether it can suppress the imbalance of homeostasis in duced by post-traumatic stress disorder. OBJECTIVE: To observe the effect of cervical sympathetic ganglia blockon the mortality of mice with combined radiation and burn injury, andwhether it can become an easy and effective method to treat secondarydamage after serious trauma. DESIGN: A randomized grouping design, an animal controlled experiment. SETTING: Department of Anesthesiology, Guangzhou General Hospital, Guangzhou Military Area Command of Chinese PLA.MATERIALS: The experiments were carried out in the Institute of Combined Injury, the Third Military Medical University of Chinese PLA between February 2004 and July 2005. Totally 160 Kunming mice were randomly divided into control group (n=50) and cervical sympathetic ganglia block group (n=50). In the control group, the mice were only induced to models of combined radiation and bum injury, and treated with injection of 0.3 mL saline at cervical part. In the cervical sympathetic ganglia block group, the mice were induced to models of combined radiation and burn injury, and then treated with cervical sympathetic ganglia block, once a day for 14 days continuously.METHODS: Methods to induce injury in the animals: ① Radiation injury: The mice were given even radiation of 60Coγ ray (5 Gy) at a distance of 1.5 m to the whole body, the rate of absorptive dosage was (5.17-5.33) mGy/s. ② Burn injury: After the radiation injury, coagulated gasoline was smeared on the back and burnt for 8 s to induce degree Ⅲ burn injury of 15% of the total body surface, which was proved by the pathological section. Methods of cervical sympathetic ganglia block: Cervical sympathetic ganglia block was given bilaterally, and then the mice were injected with 0.2 mL lidocaine (5 g/L), and it was observed whether the symptoms similar to Horner syndrome (hyperemia of conjunctiva, drooping eyelid,blushing, smaller eyeslit) occurred or not at 5 minutes after injection.MAIN OUTCOME MEASURES: The mortality at 2, 5, 7, 10, 20 and30 days after injury and the changes of the numbers of red blood cells,white blood cells and blood platelet in peripheral blood at 7, 14 and 21 days after injury were observed in both groups. The effects of cervical sympathetic ganglia block on the levels of tumor necrosis factor-alpha (TNF-α),interleukin-1β (IL-1β) and interleukin-6 (IL-6) in serum at 3, 6 and 14days after combined radiation and burn injury were also observed.RESULTS: All the 160 mice were involved in the analysis of results without deletion. ① Compared with the control group, the mortalities at 5,7, 10, 15, 20 and 30 days in the cervical sympathetic ganglia block group were significantly decreased [control group: 8%, 22%, 32%, 54%, 74%,82%, 90%; cervical sympathetic ganglia block group: 8%, 14%, 16%, 22%,28%, 34%, 56%]. ② Compared with the control group, the numbers of red blood cells, white blood cells and blood platelets in peripheral blood at 7,14 and 21 days after injury in the cervical sympathetic ganglia block group were significantly increased [at 21 days: red blood cells: 23.21×1012 L-1, 14.58×1012 L-1; blood platelet: 16.87×1011 L-1, 12.57×1011 L-1; white blood cells: 20.65×109 L-1, 14.58×109 L-1]. ③ The levels of TNF-α, IL-1β andIL-6 in serum at 3, 6 and 14 days after injury in the cervical sympathetic ganglia block group were significantly decreased as compared with those in the control group [at 14 days: TNF-α: 189, 365 ng/L; IL-1β: 14, 23 ng/L;IL-6: 70, 132 ng/L].CONCLUSION: Cervical sympathetic ganglia block can significantly decrease the mortality of animals with combined radiation and burn injury,and it is an easy and effective method to treat serious trauma, and the mechanism may be realized through accelerating the recovery of hematopoietic function and suppressing the excessive inflammatory reaction.
10.Human amniotic membrane loaded with porcine keratinocytes for constructing epidermal substitute of skin
Guohe YAN ; Yongping SU ; Feng WANG ; Guoping AI ; Tianmin CHENG ; Huaien ZHENG ; Xinze RAN ; Hong XIAO ; Chongfu TAN
Chinese Journal of Tissue Engineering Research 2005;9(22):245-247
BACKGROUND: As a kind of semitransparent membrane, human amniotic membrance contains many kinds of nutrients, which is a good biological material loaded with keratinocytes.OBJECTIVE: To construct epidermal substitute of the skin from human amniotic membrane loaded with porcine keratinocytes and examine the morphological characteristics of the growth and proliferation of keratinocytes seeded on human amniotic membrane.DESIGN: Single sample study and repetitive measured observation based on the cells.SETTING: Institute of Combined Injuries of Chinese PLA, Academy of Preventive Medicine, Third Military Medical University of Chinese PLA.MATERIALS: The experiment was completed in the State Key Laboratory of Trauma, Burn and Combined Injury and Institute of Combined Injuries of Chinese PLA, Academy of Preventive Medicine, Third Military Medical University of Chinese PLA from January to November 2001. Porcine keratinocytes was collected from Guizhou minipigs aged 3 weeks.METHODS: The primarily cultured keratinocytes of Guizhou minipigs were subcuhured, expanded and bred on the stroma surface of human amniotic membrance at the density of 1.63 × 105/cm2. The growth and proliferation of keratinocytes were observed under inverted microscope every day. From the 3rd day and the 15th day after being cultured, the growth of keratinocytes on human amniotic membrane was examined under light microscope and electron microscope.MAIN OUTCOME MEASURES: The growth of keratinocytes on human amniotic membrane was examined RESULTS: Keratinocytes evidently adhered to the stroma surface of human amniotic membrane about 30 minutes after being cultured, which was observed under inverted microscope. Most keratinocytes grew and adhered to the stroma surface of human amniotic membrane within 24 hours. Monolayer of keratinocytes formed and completely covered human amniotic membrane within 3 days. It was observed under the light microscope that the monolayer of keratinocytes adhered to human amniotic membrane and arrayed tightly. The keratinocytes presented in the shape of polygon, and plasmalemmas of keratinocytes formed many pseudopods under the observation with scanning electron microscope. Keratinocytes adhered to human amniotic membrane well and with many keratinofilaments in them under the observation with transmission electron microscope. Keratinocytes arrayed on human amniotic membrane densely with many cellular debris and some keratinocytes formed cavitations in them due to aging after growth for 15 days under the observation with inverted microscope.CONCLUSION: Human amitotic membrane is a good carrier of keratinocytes cultured on it in vitro, and is able to promote the proliferation of keratinocytes significantly. However, when keratinocytes were loaded on the human amniotic membrane for 15 days, some keratinocytes formed cavitations in them due to aging.

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