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Journal of Jilin University(Medicine Edition)
ISSN 1671-587X
CN 22-1342/R
主 任:王 丽
编 辑:姜瑾秋 李欣欣 韩宏志
    官 鑫
电 话:0431-85619279
E-mail:xuebao@jlu.edu.cn
地 址:长春市新民大街828号
    (130021)
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Current Issue
28 July 2021, Volume 47 Issue 4
Research in basic medicine
Influence of polyurethane-gel prepolymer on volume shrinkage and its improvement effect on comprehensive properties of traditional resin matrix
Gaigai YU,Huimin WANG,Zilu TIAN,Yubin YANG,Xuanyan ZHU,Song ZHU
Journal of Jilin University(Medicine Edition). 2021, 47 (4):  819-825.  DOI: 10.13481/j.1671?587X.20210401
Abstract ( 2 )   HTML ( 6 )   PDF (919KB) ( 29 )  
Objective

To synthesize an photo-active polyurethane-gel prepolymer named IBGBMA, and to explore its influence in the volume shrinkage and mechanical properties of the traditional resin matrix.

Methods

By solution polymerization,isophorone diisocyanate, bisphenol-A and hydroxyethyl methacrylate,etc.were used as the raw materials,and 1,4-butanediol and trihydroxymethylpropane were used as the chain extenders and crosslinkers, an photo-active polyurethane-gel prepolymer(IBGBMA) which could realize the chemical bond combination with the light-curable resin matrix was synthesied, and its structure was characterized by Fourier transform infrared spectroscopy(FT-IR).The Bis-GMA/TEGDMA (7∶3) resin matrix without IBGBMA was used as control group, and the matrixes added with 5%, 10%, 15% and 20% IBGBMA were used as experimental groups. The volume shrinkage rates of each group after curing were detected by the density bottle method, and the universal testing machine was used to detect the flexural strength(FS) and elastic modulus(EM), the Vickers hardness tester was used to detect the Vickers hardness, and the FT-IR peak area method was used to detect the double bonds conversion rate.

Results

The FT-IR results showed the characteristic absorption peaks of a photo-curable group (C=C) at 1 638 cm-1 as well as the group (-NHCOO-) at 3 310 cm-1, indicating that the active target product IBGBMA was synthesized,which can polymerize with the resin matrix during light curing. With the increase of IBGBMA addition amount, the volume shrinkage rates in each experimental group were decreased, and they were all lower than those in control group (P<0.05).Compared with control group,the FS and EM in 20%IBGBMA experiment group were decreased(P<0.05),but there was no statistically differences in the other experimental groups(P>0.05).The Vickers hardness in various experimental groups was increased (P<0.05).The double bond conversion rates in various experimental groups were decreased with the increase of IBGBMA addition amount, and they were all lower than that in control group(P<0.05).

Conclusion

The light-curable active polyurethane-gel prepolymer is successfully synthesized which reduces the volume shrinkage. And the volume shrinkage rate in 15%IBGBMA group is significantly reduced, the Vickers hardness is significantly increased, while the EM and FS are not affected, so the comprehensive properties are improved.

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Improvement effect of Jiedu Tongluo Decoction on myocardial fibrosis in rats through TGF-β1/Smad2/3 and ROCK pathways
Dan ZHANG,Ju HUI,Jiajuan GUO
Journal of Jilin University(Medicine Edition). 2021, 47 (4):  826-833.  DOI: 10.13481/j.1671?587X.20210402
Abstract ( 5 )   HTML ( 3 )   PDF (1582KB) ( 21 )  
Objective

To explore the effect of Jiedu Tongluo Decoction on the myocardial fibrosis in the rats, and to clarify its mechanism.

Methods

A Total of 30 SPF Wistar rats were randomly divided into normal control group, model group, captopril group,low and high doses of Jiedu Tongluo Decoction groups (n=6). The myocardial fibrosis model was established by subcutaneous injection of isoproterenol hydrochloride (5 mg·kg-1). The rats in captopril group and low and high doses of Jiedu Tongluo Decoction groups were treated with 0.005 g·kg-1·d-1 captopril and 1, 5 g·kg-1·d-1 Jiedu Tongluo Recipe for 28 d,and the rats in control group and model group were given the equal volume of distilled water by gavage. Hematoxylin-eosin (HE) staining and Masson tri-chrome staining were applied to observe the pathomorphology of myocardium tissue of the rats in various groups,and the percentages of collagen fiber areas were calculated.The hydroxyproline levels in myocardium tissue of the rats were determined by alkaline hydrolysis method, and the angiotensin Ⅱ(Ang Ⅱ) levels in myocardium tissue of the rats were detected by enzyme-linked immunosorbent assay (ELISA);the expression levels of transforming growth factor-β1 (TGF-β1) and the LIM kinase (LIMK)proteins were detected by immunohistochemistry; the expression levels of Smad2, Smad3, and Rho-associated protein kinase (ROCK) mRNA in myocardium tissue of the rats were detected by Real-time fluorescence quantitative PCR (RT-qPCR),and the expression levels of Smad2/3, phosphorylated Smad2/3 (p-Smad2/3) and ROCK proteins in myocardium tissue of the rats were analyzed by Western blotting method.

Results

The myocardium tissue showed the typical fibrosis changes 28 d after isoproterenol injection; compared with normal control group, the percentage of collagen fiber area and the levels of hydroxyproline and Ang Ⅱ in myocardium tissue of the rats in model group were significantly increased (P<0.05),the expression levels of TGF-β1,and LIMK were increased(P<0.05), the expression levels of Smad2, Smad3, and ROCK mRNA were increased(P<0.05), and the expression levels of p-Smad2/3 and ROCK proteins were significantly increased (P<0.05). Compared with model control group, the percentages of collagen fiber areas and the levels of hydroxyproline and Ang Ⅱ in myocardium tissue of the rats in captopril group and low and high doses of Jiedu Tongluo Decoction groups were significantly decreased (P<0.05),and the expression levels of TGF-β1 and LIMK proteins were decreased(P<0.05),the expression levels of Smad2, Smad3, and ROCK mRNA were decreased(P<0.05),and the expression levels of p-Smad2/3 and RCOK proteins of the rats were significantly decreased(P<0.05).There were no significant differences in the above indexes of the rats between different doses of Jiedu Tongluo Decoction groups and captopril group(P>0.05).

Conclusion

Jiedu Tongluo Decoction can inhibit the process of myocardial fibrosis by inhibiting the TGF-β1/Smad2/3 and ROCK pathway.

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Improvement effect of angiotensin(1-7) on kidney injury induced by limb ischemia-reperfusion in mice and its mechanism
Liyan ZHU,Yaoming WANG,Zheng QIN,Huanhuan ZHAO,Zengying WANG,Xiuhong YANG
Journal of Jilin University(Medicine Edition). 2021, 47 (4):  834-841.  DOI: 10.13481/j.1671?587X.20210403
Abstract ( 9 )   HTML ( 2 )   PDF (1662KB) ( 30 )  
Objective

To explore the effect of angiotensin (1-7) [Ang-(1-7)] on renal injury and inflammation in the limb ischemia-reperfusion (LIR) mice by detecting the level of Ang-(1-7) in kidey tissue and serum and the expression levels of nuclear factor-κB (NF-κB), tumor necrosis factor-α (TNF-α) and Mas receptor protein in kindey tissue of the LIR mice pretreated with Ang-(1-7).

Methods

Thirty 10-week-old C57BL/6 mice were randomly divided into control group, LIR group, and LIR+Ang-(1-7) group, with 10 mice in each group. The mice in LIR group and LIR+Ang-(1-7) group were subjected to 2 h of ischemia and 4 h of reperfusion to establish the mouse LIR models. The mice in LIR+Ang-(1-7) group were treated Ang-(1-7) (24 μg·kg-1·h-1) for two weeks before LIR by placing subcutaneous osmotic pump. The mice in LIR group were treated the same amount of normal saline for two weeks before LIR by placed subcutaneous osmotic pump. Automatic biochemical analyzer was used to determine the levels of serum creatinine (SCr) and urea nitrogen (BUN) of the mice; HE staining was used to observe the pathomorphology of kidney tissue of the mice, and renal tubular pathological score was performed. Enzyme linked immunosorbent assay (ELISA) was used to determine the Ang-(1-7) levels in kidney tissue and serum of the mice. Immunohistochemical staining and Western blotting methods were used to detect the expression, distribution and the expression levels of NF-κB,TNF-α and Mas receptor proteins in the mouse kidney tissue.

Results

Compared with control group, the levels of serum SCr and BUN of the mice in LIR group were increased significantly (P<0.05), and the obvious pathological damages such as hyperemia, renal tubular dilation, edema, necrosis, and inflammatory cell infiltration were observed in the kidney tissue; the pathological injury score of kidney tissue was increased(P<0.05); the Ang-(1-7) levels in kidney tissue and serum of the mice were decreased significantly (P<0.05), and the expression levels of NF-κB, TNF-α and Mas receptor proteins were increased significantly (P<0.05). Compared with LIR group, the levels of serum SCr and BUN of the mice in LIR+Ang-(1-7) group were decreased significantly (P<0.05),and the kidney congestion and other damages were significantly reduced;the pathological injury score of kindey tissue was decreased(P<0.05); the Ang-(1-7) levels in kidney tissue and serum of the mice were increased significantly (P<0.05); the expression levels of NF-κB and TNF-α proteins were decreased (P<0.05), and the expression level of Mas receptor protein in kidney tissue of the mice in LIR+Ang-(1-7) group was increased significantly (P<0.05).

Conclusion

Mouse LIR can induce the distal kidney damage, and Ang-(1-7) may reduce the expression levels of NF-κB and TNF-α proteins in kidney tissue to improve the kidney damage caused by mouse LIR.

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Effect of abdominal manipulation on remodeling of hippocampal neurons in chronic stress-induced chronic fatigue syndrome and its mechanism of negative feedback regulation of hippocampus-HPA axis
Mingzhu PAN,Jian LI,Bing RONG,Jun JIA,Huanan LI
Journal of Jilin University(Medicine Edition). 2021, 47 (4):  842-848.  DOI: 10.13481/j.1671?587X.20210404
Abstract ( 13 )   HTML ( 4 )   PDF (1786KB) ( 4 )  
Objective

To observe the effect of abdominal manipulation on the remodeling of hippocampal neurons in the model rats with chronic stress-induced chronic fatigue syndrome (CFS), and to analyze the mechanism of negative feedback regulation of hippocampus-hypothalamic-pituitary-adrenal axis(hippocampus-HPA axis).

Methods

A total of 60 healthy adult female Wistar rats were selected and divided into normal group, model group and experimental group, there were 20 rats in each group. The rats in model group and experimental group were used to establish the CFS models by multiple stress, and abdominal manipulation intervention was performed in experiment group after model establishment, while the rats in normal group received no treatment.The behavior effect indicators were used to verify the models; the urtramicromorphology of hippocampal neurons of the rats was observed by electron microscope. The positive expression rates of FK506 binding protein (FKBPs),glucocorticoid receptor (GR) and N-methyl-D-aspartate receptor (NMDAR) were detected by immunohistochemical method, and the serum levels of adrenocorticotropic hormone (ACTH),cortisol(CORT),corticotropin releasing hormone (CRH) and glucocorticoid (GC) of the rats were detected by ELISA method.

Results

The behavior effect indicator examination results showed the time for immobility of the rats in model group and experiment group before abdominal manipulation was longer than that in normal group(P<0.05),and the time for exhaustive swimming and the speeds of horizontal movement of the mice in model group and experiment group were lower than those in normal group(P<0.05);compared with model group, the time for immobility of the rats in experiment group was significantly shortened(P<0.05),the exhaustive swimming time was significantly prolonged(P<0.05),and the horizontal movement speed was significantly increased(P<0.05).Compared wtih normal group,shrinkage of hippocampal neurons, some invaginated neurons and apoptotic bodies were observed in model group under electron microscope;compared with model group, the hippocampal neurons in experiment group were significantly alleviated. Compared with normal group, the positive expression rates of FKBPs, GR and NMDAR in the hippocampus tissue of the rats in model group and experiment group were significantly decreased(P<0.05); compared with model group, the positive expression rates of FKBPs, GR and NMDAR in the hippocampus tissue of the rats in experiment group were significantly increased(P<0.05). Compared with normal group, the serum levels of ACTH, CORT, CRH and GC of the rats in model group were significantly increased(P<0.05); compared with model group, the serum levels of ACTH, CORT, CRH and GC of the rats in experiment group were significantly decreased(P<0.05).

Conclusion

Abdominal manipulation can promote the remodeling of damaged hippocampal neurons caused by stress response, reduce the levels of ACTH, CORT and CRH, and maintain the negative feedback balance of hippocampus-HPA axis through regulating FKBPs-GR-NMDAR pathway.

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Effect of asparagine synthetase on reduction of skeletal muscle in APP/PS1 mice transplanted with melanoma
Ying DONG,Hang LYU,Jianv GUO,Hong ZHENG,Dan GUO,Like WANG,Dan WANG,Changjie CHEN,Liankun SUN,Yong ZHANG,Chunyan YU
Journal of Jilin University(Medicine Edition). 2021, 47 (4):  849-856.  DOI: 10.13481/j.1671?587X.20210405
Abstract ( 9 )   HTML ( 2 )   PDF (1180KB) ( 12 )  
Objective

To investigate the role of asparagine synthetase(ASNS) on reduction of skeletal muscle in the amyloid precursor protein/presenilin 1(APP/PS1) mice transplanted with melanoma,and to clarify its mechanism.

Methods

The C57BL/6J(C57) and APP/PS1 mice were transplarted with melanoma cells (C57 transplanted tumor group and APP/PS1 transplanted tumor group),at the same time, C57 control group and APP/PS1 control group were set up.The mass of gastrocnemius was measured and the sarcopenia index (SI) was calculated with the gastrocnemius mass to body weight ratio.The morphology of gastrocnemius tissue of the mice was determined by HE staining. The mRNA expression levels of muscle specific atrophy gene E3 ubiquitin protein ligase 1 (Atrogin-1),muscle ring finger protein 1(MuRF1) and ASNS in gastrocnemius tissue of the mice were measured by Real-time fluorescence quantitative PCR(RT-qPCR). The expression levels of ASNS and endoplasmic stress related glucose regulated protein 78(GRP78),activating transcription factor-4(ATF4),eukaryotic cell initiation factor 2α(eIF2α),phosphorylated eIF2α(p-eIF2α) in gastrocnemius tissue of the mice were measured by Western blotting method.

Results

Compared with C57 control group, the SI of the mice in C57 transplanted tumor group was decreased (P<0.05),gastrocnemius cells had hyaline degeneration,the sarcoplasm was stained blue, and some muscle fiber showed intranuclear migration; the expression levels of Atrogin-1,MuRF1 and ASNS mRNA in gastrocnemius tissue of the mice were increased (P<0.05), and the expression level of ASNS protein was increased (P<0.05);the expression levels of GRP78 and ATF4 and the ratio of p-eIF2α/eIF2α were increased(P<0.05).Compared with APP/PS1 control group,the SI of the mice in APP/PS1 transplanted tumor group was decreased (P<0.05), some muscle fibers of gastrocnemius were swelling, and some were thin; intranuclear migration was easily seen, and the sarcoplasm was red; the expression levels of Atrogin-1,MuRF1 and ASNS mRNA in gastrocnemius tissue were increased (P<0.05),and the expression level of ASNS protein was increased (P<0.05);the expression levels of GPR78 and ATF4 proteins and the ratio of p-eIF2α/eIF2α were increased(P<0.05).Compared with C57 transplanted tumor group,the SI of the mice in APP/PS1 transplanted tumor group was decreased (P<0.05),the level of Atrogin-1 mRNA was increased (P<0.05),and the level of ASNS mRNA in gastrocnemius tissue was decreased (P<0.05); there was no difference in the expression level of ASNS protein(P>0.05);the expression levels of GRP78 and ATF4 proteins and the ratio of p-eIF2α/eIF2α were decreased (P<0.05).

Conclusion

Gastrocnemius in the APP/PS1 transplanted melanoma mice is more prone to reduction of skeletal muscle, and its mechanism may be associated with the reduced levels of adaptive response in ASNS transcription, and endoplasmic reticulum stress in PERK-eIF2a-ATF4 signaling pathway may be involved in the regulation of ASNS transcriptional activity.

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Therapeutic effect of estradiol combined with 1,25-dihydroxyvitamin D3 on postmenopausal osteoporosis in rats
Xining LI,Wei WENG,Zheyuan SHEN,Xiaojie DOU,Jikang MIN
Journal of Jilin University(Medicine Edition). 2021, 47 (4):  857-864.  DOI: 10.13481/j.1671?587X.20210406
Abstract ( 4 )   HTML ( 2 )   PDF (1333KB) ( 5 )  
Objective

To investigate the effect of estradiol combined with 1, 25-dihydroxyvitamin D3[1,25(OH)2D3] in the postmenopausal osteoporosis (PMOP) rats, and to clarify its mechanism.

Methods

Forty female SD rats were randomly divided into control group, osteoporosis model group (model group),ovarian castratiom and estradiol intervention group (estradiol group),ovarian castration and estradiol combined with 1, 25-(OH)2D3 group (estradiol combined with 1, 25 (OH)2D3 group);there were 10 rats in each group.The serum of rats were collected after 24 weeks of feeding, and the levels of alkaline phosphatase (ALP), osteocalcin (OC),osteoprotectin (OPG),amino-end peptide of type Ⅰ procollagen (PINP),tartrate-resistant acid phosphatase (TRACP) and nuclear factor kappa B receptor activator ligand (RANKL) were detected by ELISA method.Bilateral femurs were taken and the changes of bone tissue were observed by HE and TRAP staining. The elastic modulus, stiffness, maximum stress, maximum lord and bone mineral density (BMD) of bone tissue of the rats in each group were detected by biomechanics and BMD. The mRNA and protein expression levels of ALP, OC, OPG and RANKL in bone tissue of the rats in various groups were detected by RT-PCR and Western blotting methods.

Results

Compared with control group,the levels of serum ALP,OC,OPG,PINP,TRACP and RANKL were significantly increased(P<0.01),the periosteum thickness was increased,the number of osteoclasts was increased,and a little of punctate inset-etched defect was found under periosteum,presenting the typical features of osteoporosis; the biomechanical index and BDM of femurs were decreased(P<0.01),the expression levels of ALP,OC,OPG,and RANKL mRNA and proteins in bone tissue were significantly increased(P<0.01).Compared with model group,the levels of serum ALP,PINP,TRACP,and RANKL of the rats in estradiol combined with 1,25(OH)2D3 group were significantly decreased(P<0.01),the ratio of OPG/RANKL was increased(P<0.01),the osteoclasts in periosteum were proliferated actively,and there was no punctate inset-etched defect;the BMD of femurs of the rats and the biomechrical indexes were significantly increased(P<0.01),and the expression levels of ALP,OC and RANKL mRNA and proteins in bone tissue of the rats were decreased(P<0.01).Compared with estradiol group,the levels of serum OC and OPG of the rats in estradiol combined with 1,25(OH)2D3 group and the ratio of OPG/RANK were increased(P<0.01),the levels of TRACP and RNAKL were significantly decreased(P<0.01),the BMD of femurs and the biomechical indexes elastic modulus and maximum stress were significantly increased(P<0.01),the expression levels of OPG mRNA and protein in bone tissue were increased(P<0.01),and the expression levels of RANKL mRNA and protein were significantly decreased(P<0.01).

Conclusion

Estradiol combined with 1,25(OH)2D3 is more effective in promoting the proliferation and differentiation of osteoblasts, and the combination can reduce the adverse reactions caused by excessive hormone replacement while treating PMOP.

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Effect of Dushen Tang on cognitive dysfunction of aging model rats based on cAMP/PKA/CREB signal pathway
Jifeng WANG,Xiaoran LIU,Xin SUI,Mo KAN,Hui LI,Wenjun GUO,Qing YANG,Zhuang ZHANG,Sitong MING,Na LI,Xiaobo QU
Journal of Jilin University(Medicine Edition). 2021, 47 (4):  865-873.  DOI: 10.13481/j.1671?587X.20210407
Abstract ( 5 )   HTML ( 2 )   PDF (2196KB) ( 8 )  
Objective

To explore the effect of Dushen Tang(DST) on the neuronal damage, the superoxide dismutase (SOD) and glutathione (GSH) activities in brain tissue, the malondialdehyde (MDA) and cyclic adenosine monophosphate (cAMP) levels in the D-galactose (D-gal)-induced aging model rats and the activation of cyclic adenosine monophosphate/protein kinase A/CAMP response element binding protein (cAMP/PKA/CREB) signal pathway, and to elucidate the improvement effect of DST on the cognitive dysfunction of the D-gal-induced aging model rats.

Methods

Forty SD rats were randomly divided into control group, model group, positive drug group, and DST group, with 10 rats in each group. Except for control group, the rats in other groups were given intraperitoneal injection of D-gal at a dose of 500 mg·kg-1·d-1 for 40 d to establish the aging models. On the 5th day of the model construction, the rats in positive drug group was given vitamin E 0.027 g·kg-1, and the rats in DST group were given DST 5mL (equivalent to 0.3 g·kg-1·d-1 crude drug), for 36 d. The states of the rats were observed and the growth curves were draw; the water maze was used to detect the learning and memory abilities of the rats; the biochemical and ELISA kits were used to detect the activities of SOD, GSH and the levels of MDA and cAMP in the rat brain tissue; the transmission electron microscope was used to observe the ultrastructures of the rat hippocampus neurons; Congo red staining was used to observe the age spots in hippocampus tissue of the rats; Western blotting was used to detect the expression levels of cAMP/PKA/CREB signaling pathway-related proteins in hippocampus tissue of the rats.

Results

Compared with model group, the body weights of rats in positive drug group and DST group were increased, the learning and memory abilities were increased (P<0.05), the activities of SOD and GSH and the levels of cAMP in brain tissue of the rats were significantly increased(P<0.05),and the levels of MDA were decreased (P<0.05). Compared with model group, the pathological changes of neuron ultrastructures of the rats in positive drug group and DST group were improved and the areas of orange-red sediments and age spots in hippocampus tissue of the aging model rats were significantly decreased(P<0.01);the expression levels of PKA, cAMP, adenylate cyclase(ADCY),CREB,and brain-derived neurotrophic factor (BDNF) proteins in brain tissue of the rats were significantly increased(P<0.05),and the expression levels of guanine nucleotide-binding protein alpha inhibitor(GNAI) were decreased (P<0.05).

Conclusion

DST can significantly improve the cognitive ability of the D-gal-induced aging model rats,and its mechanism may be related to scavenging free radicals and activating cAMP/PKA/CREB signaling pathway to improve the nerve cell damage.

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Induction of 3C protein of Coxsackievirus A16 in apoptosis of human rhabdomyosarcoma cells and its mechanism
Yingying SHI,Shitong CHEN,Bo HU,Duo SHEN,Kuang ZHU,Siwei YE,Jinmei FENG
Journal of Jilin University(Medicine Edition). 2021, 47 (4):  874-881.  DOI: 10.13481/j.1671?587X.20210408
Abstract ( 3 )   HTML ( 1 )   PDF (1162KB) ( 2 )  
Objective

To investigate the effect of Coxsackievirus A16 3C protein on the apoptosis of rhabdomyosarcoma RD cells, and to illuminate its possible mechanism.

Methods

The RD cells were cultured in vitro and divided into control group (transfected with 0.4 μg pCMV-HA), 0.1 μg pCMV-HA-3C group (transfected with 0.1 μg pCMV-HA-3C and 0.3 μg pCMV-HA), 0.2 μg pCMV-HA-3C group (transfected with 0.2 μg pCMV-HA-3C and 0.2 μg pCMV-HA),0.4 μg pCMV-HA-3C group (transfected with 0.4 μg pCMV-HA-3C) and protein kinase B(AKT) activator SC79 treatment group (pretreated with 4 mg·L-1 SC79 for 4 h, then transfected with 0.2 μg pCMV-HA-3C and 0.2 μg pCMV-HA).The survival rates of cells in various groups were detected by MTT assay,the apoptotic rates were detected by flow cytometry, the expression levels of Bcl-2 homologous antagonist /killer (Bak),Bcl-2 associated agonist of cell death (Bad) and p53 up-regulated modulator of apoptosis (PUMA) mRNA were detected by Real-time fluorescence quantitative PCR (RT-qPCR),and the expression levels of Bad, Bak, PUMA, PARP, cleaved-PARP, cleaved-caspase-3 and phosphorylated AKT (p-AKT) proteins in the RD cells in various groups were detected by Western blotting method.

Results

Compared with control group, the survival rate of the cells 0.4 μg pCMV-HA-3C group was significantly decreased (P<0.01). Compared with control group, the apoptotic rate of the cells in pCMV-HA-3C group was significantly increased (P<0.01). Compared with control group,the mRNA and protein expression levels of Bak, Bad and PUMA in 0.2 μg PCMV-HA-3C group were significantly increased (P<0.05), and the expression levels of cleaved-PARP and cleaved-caspase-3 proteins in different doses of pCMV-HA-3C groups were significantly increased (P<0.05).Compared with control group, the expression levels of cleaved-PARP and cleaved-caspase-3 proteins in 0.2 μg pCMV-HA-3C group were significantly increased (P<0.05),and the expression level of p-AKT protein was significantly decreased (P<0.01). Compared with 0.2 μg pCMV-HA-3C group,the the expression levels of cleaved-PARP,cleaved-caspase-3 and AKT proteins in the cells in SC79 treatment group were significantly decreased(P<0.05),and the p-AKT protein expression level was significantly increased (P<0.01).

Conclusion

Coxsackievirus A16 3C protein may induce the apoptosis of RD cells by inhibiting AKT signaling pathway.

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Effect of combination of natural sounds during infancy on anxiety-like behaviors in adult rats
Qian WANG,Siyue ZHUANG,Guilong WU,Shengtian LI
Journal of Jilin University(Medicine Edition). 2021, 47 (4):  882-887.  DOI: 10.13481/j.1671?587X.20210409
Abstract ( 4 )   HTML ( 1 )   PDF (999KB) ( 4 )  
Objective

To clarify the effect of combined natural sound intervention during infancy on the anxiety-like behaviors in the adult rats by testing the offspring male SD rats’ anxiety-like behaviors.

Methods

Six SD rats with 14-15 d of gestation were randomly divided into standard noise control group (NN group),Mozart K448 group (MK group) and combination of natural sound group (NS group). The rats in NN group were not given any special sound stimulation,the rats in MK group were given 2 h of MK sound stimulation at a volume of 50-70 dB per day for 30 d,and the rats in NS group were given a combination of natural sounds at a volume of 65-75 dB for 2 h a day for 30 d. Five weeks after the end of stimulation, 11 male rats with the similar body weight in NN group and in NS group, and 8 rats in MK group were selected for open field experiment,light-dark transition test and elevated plus-maze test.The open field experiment was used to analyze the total movement distance, rearing times, the number of entering the central area, and the time of entering the central area. The percentage of time staying in light box and the number of transition between light and dark boxes and the total movement distance of rats were analyzed in the light-dark transition test. The elevated plus-maze test was used to analyze the percentage of time in open arms, the number of open arms entries and the distance of open arms.

Results

In the open field experiment, there were no statistically significant differences among three groups in the total movement distance, rearing times, the time of entering the central area, and the number of entering the central area (P>0.05). The results of light-dark transition test showed that the time in the light box of the rats in NS group was significantly longer than that in NN group (P<0.05); the number of transition between light and dark boxes of the rats in NS group was also significantly increased (P<0.05 or P<0.01). The results of elevated plus-maze test also showed that the percentage of time in open-arms of the rats in NS group was significantly longer than that in NN group (P<0.05).

Conclusion

Intervention of natural sounds during infancy can reduce the anxiety-like behaviors in the mature male SD rats.

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Regulatory effect of glucose-regulating protein 78 on radiotherapy sensitivity of liver cancer cells and its mechanism
Jie YANG,Wubin HE,Ni AN,Wencong KONG,Rongjian SU,Xuezhe WANG
Journal of Jilin University(Medicine Edition). 2021, 47 (4):  888-895.  DOI: 10.13481/j.1671?587X.20210410
Abstract ( 13 )   HTML ( 3 )   PDF (1286KB) ( 3 )  
Objective

To investigate the effect of glucose-regulating protein 78 (GRP78) on the radiotherapy sensitivity of liver cancer cells, and to preliminarily explore its molecular mechanism.

Methods

Real-time fluorescence quantitative PCR(RT-qPCR) and Western blotting methods were used to detect the expression levels of GRP78 mRNA and protein in the liver cancer SMMC-7721, HepG2, PLC, Huh7 and QGY-7703 cells, then the high and low GRP78 expression liver cells were selected as the research objects. GRP78 was targeted silenced by RNA interference technique in highly expressed human liver cancer SMMC-7721 cells, which was named siGRP78-SMMC-7721(siGRP78 group), and siNC-SMMC-7721 was used as control group(siNC group). GRP78 was over-expressed in the low-expression liver cancer HepG2 cells by gene recombination technique, which was named Flag-GRP78-HepG2(Flag-GRP78 group),and empty vector 3×Flag-HepG2 was used as control group(3×Flag group).After irradiation with 0, 2, 4, 6, 8 and 10 Gy X-ray, the survival rates of liver cancer cells in various groups were determined by MTT assay. The proliferation rates of liver cancer cells in various groups were determined by EdU fluorescence staining. Western blotting method was used to detect the expression levels of phospatidylinositol-3(PI3K)/protein kinase B(Akt) signaling pathway proteins.

Results

The results of RT-qPCR and Western blotting methods showed that the expression levels of GRP78 mRNA and protein were the highest in SMMC-7721 cells and the lowest in HepG2 cells,so the SMMC-7721 and HepG2 cells were selected as the subjects.The MTT assay results showed that the survival rates of liver cancer cells were decreased with the increase of radiation dose;compared with siNC group, the survival rates of cells in siGRP78 group were significantly reduced after X-ray irradiation above 6 Gy(P<0.01);compared with 3×Flag group, the survival rates of cells in Flag-GRP78 group were significantly increased after X-ray irradiation above 4 Gy (P<0.05 or P<0.01).The EdU fluorescence staining results showed that compared with siNC group, the proliferation rate of cells in siGRP78 group was significantly decreased (P<0.05);compared with 3×Flag group, the cell proliferation rate in Flag-GRP78 group was significantly increased (P<0.05).The results of Western blotting method showed that compared with siNC group, the expression levels of phosphorylated PI3K(p-PI3K)and phosphorylated Akt(p-Akt) proteins in the cells in siGRP78 group were significantly decreased(P<0.01). Compared with 3×Flag group, the expression levels of p-PI3k and p-Akt in the cells in Flag-GRP78 group were significantly increased(P<0.01).

Conclusion

High expression GRP78 can decrease the radiosensitivity of liver cancer cells, and its mechanism may be related to the activation of PI3K/Akt signaling pathway.

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Effects of targeting-silenced ATRX on cell cycle arrest and apoptosis induced by radiation in cervical cancer HeLa cells
Lijing QIN,Bing HAN,Zhongqi LI,Qi SUO,Zhen JIA,Hongli CUI,Weiqiang XU,Fang FANG,Zhicheng WANG
Journal of Jilin University(Medicine Edition). 2021, 47 (4):  904-910.  DOI: 10.13481/j.1671?587X.20210412
Abstract ( 2 )   HTML ( 3 )   PDF (1245KB) ( 7 )  
Objective

To targetedly silence the α-thalassemia/mental retardation syndrome X-linked (ATRX) in the cervical cancer HeLa cells, and to explore the roles of targeting-silenced ATRX in radiation-induced cell cycle arrest and apoptosis.

Methods

The HeLa cell models stably silenced ATRX were established by RNA interference (RNAi) technique, which was named shATRX1-HeLa, and shCon-HeLa was used as negative control.After 0, 2 and 8 Gy X-ray irradiation, the cell proliferation activities in various groups were detected by CCK-8 method, and the percentages of HeLa cell in the G0/G1, S and G2/M phases in various groups were measured by PI staining and flow cytometry. In addition, the apoptotic rates were measured by AnnexinⅤ-PE/7-AAD double staining and flow cytometry, and the expression levels of poly[ADP-ribose] polymerase 1(PARP1),cleaved caspase-9 and cleaved caspase-3 proteins in the cells in various groups were detected by Western blotting method.

Results

At 0, 6, 10, 24 and 48 h after 0, 2 and 8 G y X-ray irradiation, compared with shCon-HeLa group, the proliferation activities of the cells in shATRX1-HeLa group were decreased significantly (P<0.05 or P<0.01). Compared with shCon-HeLa group, after 0 and 2 Gy X-ray irradiation, the percentages of cells in G0/G1 phase in shATRX1-HeLa group were decreased significantly(P<0.05), and the percentages of cells in G2/M phase and the apoptotic rates were increased significantly (P<0.05 or P<0.01). Compared with shCon-HeLa group, after 8 Gy X-ray irradiation, the percentage of cells in G0/G1 phase and the apoptotic rate were increased significantly (P<0.05), and the percentage of cells in G2/M phase was decreased significantly (P<0.05).The Western blotting results showed that compared with shCon-HeLa group, at 24 and 48 h after 2 and 8 Gy X-ray irradiation, the expression amounts of PARP1, cleaved caspase-9 and cleaved caspase-3 proteins in the cells in shATRX1-HeLa group were increased.

Conclusion

Targeting-silenced ATRX can decrease the G2/M phase arrest of the cells induced by radiaton in the HeLa cells, and inhibit the cell proliferation and promote the apoptosis; the results of this study provides a new molecular target for tumor radiotherapy.

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Effect of high expression of Snail on invasion capacity of placental trophoblasts in preeclampsia rats and its mechanism
Changcun ZHANG,Aiping ZHANG,Yuqin LI
Journal of Jilin University(Medicine Edition). 2021, 47 (4):  911-918.  DOI: 10.13481/j.1671?587X.20210413
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Objective

To study the effect of high expression of zinc finger transcription factor Snail on the invasion capacity of placental trophoblasts in the preeclampsia (PE) rats, and to explore the related mechanisms.

Methods

The PE rat models were established and used as model group; the healthy rats were selected and used as control group. The primary trophoblasts were isolated and cultured from the chorionic trophoblast tissue of the PE rats. The trophoblasts in the logarithmic growth phase were selected as Neo group (transfected with pL-tdTomato-Neo plasmid) and mSnail group (transfected with pL-tdTomato-mSnail plasmid), and the untreated cells were used as blank control group. Real-time fluorescence quantitative PCR (RT-qPCR) and Western blotting methods were used to detect the expression levels of Snail mRNA and protein in chorionic trophoblast tissue of the rats in control group and model group and in the cells in blank control group, Neo group and mSnail group. Transwell chamber experiment was used to determine the cell invasion abilities in blank control group, Neo group and mSnail group. Western blotting method was used to detect the expression levels of epithelial cadherin (E-cadherin), neurogenic cadherin (N-cadherin) and Vimentin proteins in the cells.

Results

Compared with control group, the expression levels of Snail mRNA and protein in the placenta chorionic trophoblast tissue of the rats in model group were decreased (P<0.05);under fluorescence microscope, the transfection efficiencies in Neo group and mSnail group were both >80%. Compared with blank control group and Neo group, the expression levels of Snail mRNA and protein in the cells in mSnail group were increased(P<0.05), the average number of cells passing through the micropore in each field of view was increased(P<0.05), the expression level of E-cadherin protein was decreased (P<0.05),and the expression levels of N-cadherin and Vimentin proteins were increased(P<0.05). Conclusion High expression of Snail can enhance the invasion ability of placental trophoblasts in the PE rats and its mechanism may be related to Snail’s promotion of the epithelial-mesenchymal transition of placental trophoblasts.

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Neuroprotective effect of Withaferin A on ischemic stroke rats and its mechanism
Dimi ZHOU,Lu GAN,Lin CHEN,Chengfang ZHOU,Liang ZENG
Journal of Jilin University(Medicine Edition). 2021, 47 (4):  919-925.  DOI: 10.13481/j.1671?587X.20210414
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Objective

To investigate the protective effect of Withaferin A on the neurological function of the rats with ischemic stroke, and to clarify its possible mechanism.

Methods

A total of 75 SD rats were randomly divided into sham operation group, model group, positive control group (nimodipine group,12 mg·kg-1),low dose of Withaferin A group (25 mg·kg-1) and high dose of Withaferin A group (100 mg·kg-1),and there were 15 rats in each group. The rat models of ischemic stroke were established by improved Longa line embolization. After successful modeling, the drugs were administered by gavage once per day for 14 d. Subsequently, the neurological function, the content of water in brain and the volume of cerebral infarction were evaluated. The activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), catalase (CAT) and the levels of malondialdehyde (MDA) in brain tissue of the rats were determined by biochemical method. The apoptotic rates of cells of the rats were detected by TUNEL staining, and the mRNA and protein expression levels of nuclear E2-related factors 2 (Nrf2) and heme oxygenase-1 (HO-1) in brain tissue of the rats were respectively detected by RT-qPCR and Western blotting methods.

Results

Compared with sham operation group, the score of neurological function, the volume of cerebral infarction, the content of water in brain tissue, the apoptotic rate of cells and the level of MDA in brain tissue of the rats in model group were significantly increased (P<0.05), while the activities of SOD, GSH-PX and CAT, as well as the mRNA and protein expression levels of Nrf2 and HO-1 in brain tissue of the rats were significantly decreased (P<0.05). Compared with model group, the scores of neurological function, the volumes of cerebral infarction, the contents of water in brain tissue, the apoptotic rates of cells and the levels of MDA in brain tissue of the rats in high dose of Withaferin A group and nimodipine group were significantly reduced (P<0.05), and the activities of SOD, GSH-PX and CAT as well as the mRNA and protein expression levels of Nrf2 and HO-1 in brain tissue of the rats were significantly increased (P<0.05); However, there were no significant differences in the related indexes in low dose of Withaferin A group (P>0.05).

Conclusion

Withaferin A can improve the level of antioxidant in the rats with ischemic stroke, reduce the oxidative damage of brain tissue, inhibit the apoptosis of nerve cells of brain tissue, and improve the neurological function of the rats; its mechanism may be related to the activation of Nrf2/HO-1 signaling pathway.

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Effect of chloroquine on gemcitabine-resistant cells by affecting autophagy and mitochondrial function of pancreatic cancer cells and its mechanism
Lu LU,Dongming LI,Xueguo WANG,Dan SONG,Taicheng WANG,Hongyan ZHAO,Xiaoyong WU
Journal of Jilin University(Medicine Edition). 2021, 47 (4):  926-933.  DOI: 10.13481/j.1671?587X.20210415
Abstract ( 5 )   PDF (1479KB) ( 6 )  
Objective

To investigate the effect of chloroquine (CQ) on the gemcitabine(GEM) resistant pancreatic cancer(PC) cells, and to clarify its related mechanism;

Methods

The GEM resistant PANC1 cell line (PANC1/GEM) was established by low concentration sequential increasing method, and the cells were divided into GEM resistant group (PANC1/GEM group), CQ group and GEM resistant+CQ group (PANC1/GEM+CQ group), and the PANC1 cells were used as control group at the same time. CCK-8 method was used to verify whether the cell line was successfully established.The number of autophagy bodies was observed by transmission electron microscope, and the expression levels of autophagy marker proteins cytoplas microtubule-associated protein 1 light chain 3 (LC3-Ⅰ),membrane microtubule-associated protein 1 light chain 3 (LC3-Ⅱ) and P62 proteins were detected by Western blotting method; JC-1 method, DCFH-DA, Annexin Ⅴ-FITC/PI double staining and flow cytometry were used to detect the mitochondrial membrane potential,the reactive oxygen species (ROS) levels and the apoptotic rates of the cells in various groups. The expression levels of apoptosis related proteins B cell lymphoma-2(Bcl-2), Bcl-2 associated X protein(Bax) and Cleaved caspase-3 proteins in the cells in various groups were detected by Western blotting method.

Results

The PANC1/GEM cell line was successfully established, and the IC50 value of GEM to the PANC1/GEM cells was significantly increased compared with PANC1 cells(P<0.01). Compared with control group, the number of autophagy bodies and the LC3-Ⅱ/LC3-Ⅰ ratios in PANC1/GEM group, CQ group and PANC1/GEM+CQ group were significantly increased (P<0.05), and the levels of P62 protein in PANC1/GEM group and PANC1/GEM+CQ group were significantly decreased (P < 0.05), and the expression level of P62 protein in CQ group was significantly increased(P<0.05). Compared with PANC1/GEM group, the LC3-Ⅱ/LC3-Ⅰ ratio in PANC1/GEM+CQ group was significantly decreased(P<0.05), and the expression level of P62 protein was significantly increased (P<0.05).Compared with control group, the mitochondrial membrane potentials of the cells in PANC1/GEM group and PANC1/GEM+CQ group, the ROS levels and the apoptotic rates were significantly decreased (P<0.05),and the mitochondrial membrane potential,the ROS levels and the apoptotic rate in CQ group were significantly increased (P<0.05);compared with PANC1/GEM group,the mitochondrial membrane potential,the ROS level and the apoptotic rate in PANC1/GEM+CQ group were significantly increased(P<0.05).Compared with control group, the expression levels of Bcl-2 protein in PANC1/GEM and PANC1/GEM+CQ groups were increased (P<0.05), while the expression levels of Bax and Cleaved caspase-3 proteins were significantly decreased (P<0.05);the expression level of Bcl-2 protein in CQ group was significantly decreased (P<0.05),and the expression levels of Bax and Cleaved caspase-3 proteins were significantly increased (P<0.05). Compared with PANC1/GEM group, the expression level of Bcl-2 protein in PANC1/GEM+CQ group was significantly decreased (P<0.05), while the expression levels of Bax and Cleaved caspase-3 proteins were significantly increased (P<0.05).

Conclusion

CQ can significantly promote the sensitivity of drug-resistant PC cells to GEM, the mechanism of which may be related to the inhibition of autophagy of pancreatic cancer cells, the reduction of mitochondrial membrane potential, the promotion of ROS production in the cells, and the up-regulation of GEM-induced apoptosis.

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Analysis on expression characteristics of non-structural protein μNS and σNS of Nelson Bay orthoreovirus
Tengfei LI,Shiyu WANG,Xinru JIANG,Miao SUN,Tingting LI,Jiafeng LIN,Ying WANG,Yonggang LI,Xiaoli TAO
Journal of Jilin University(Medicine Edition). 2021, 47 (4):  934-942.  DOI: 10.13481/j.1671?587X.20210416
Abstract ( 1 )   HTML ( 2 )   PDF (1475KB) ( 5 )  
Objective

To detect the interaction between non-structural proteins μNS and σNS and their binding regions,and to clarify the pathogenic mechanism of Nelson Bay orthoreovirus (NBV).

Methods

The pCAG M3 and pEFHAσNS plasmids were transfected into the BHK cells separately and co-transfected with Lipo3000 transfection reagent. Immunofluorescence method was used to detect the expressions and location of μNS and σNS proteins in the cells,NBV was used to infect the BHK cells, and Western blotting method was used to detect the expressions of μNS and σNS proteins in the transfected cells.A plasmid with 10-60 amino acid(aa) residues deleted from the N-terminal of σNS protein was constructed.Immunofluorescence method was used to detect the co-localized binding region of μNS and σNS proteins in the cells. Yeast two-hybrid experiment was used to verify the binding region of μNS and σNS protein interaction.

Results

The subcellular location results showed that the μNS protein of NBV formed an inclusion body structure in the absence of other viral proteins, while the σNS protein was diffusely distributed throughout the cytoplasm.The Western blotting results confirmed that μNS and σNS proteins were expressed in the infected cells. Immunostaining was carried out with the conjugated antibody. It was observed under confocal microscope that the μNS and σNS proteins co-localized in the cytoplasm. The results of yeast two-hybrid experiment showed that the basic region of the σNS protein interacting with the μNS protein was located at 60 aa in the N-terminal region of the σNS protein within the residue region.

Conclusion

NBV autophagy-related protein σNS can be co-expressed in the viral inclusion bodies by interacting with μNS protein. The binding region of σNS and μNS protein is located within the 60 aa residue region of the N-terminal of σNS protein.

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Improvement effect of diosgenin on symptoms of synovitis rats and its reglatory effect on TLR2-NF-κB signaling pathway and mechanism
Haojie WU,Minghui ZHANG,Chengzhi HONG
Journal of Jilin University(Medicine Edition). 2021, 47 (4):  943-950.  DOI: 10.13481/j.1671?587X.20210417
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Objective

To investigate the improvement effect of diosgenin on the symptoms of synovitis rats and the influence in Toll-like receptor 2 (TLR2) - nuclear factor kappaB(NF-κB) signaling pathway, and to elucidate the possible mechanism of diosgenin in the treatment of synovitis.

Methods

The synovitis models were established by intradermal injection of Freund’s complete adjuvant,and the model rats were randomly divided into model group(n=10),positive drug group (indomethacin 10 mg·kg-1)(n=11), low dose of diosgenin group (60 mg·kg-1)(n=12), and high dose of diosgenin group (120 mg·kg-1)(n=12); another 10 healthy rats were set as control group. Joint pain scoreing and water volume methods were used to evaluate the changes of joint pain and swelling on the 3rd, 7th, 14th and 21st days after intervention. The levels of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in synovial tissue of the rats in various groups were detected by enzyme-linked immunosorbent assay method. HE staining was used to observe the pathomorphology of synovium tissue of the rats in various groups. Western bloting method was used to detect the expression levels of TLR2, MyD88, NF-κB and p-NF-κB p65 proteins in synovium tissue of the rats in various groups.

Results

Compared with control group, the pain scores of the rats in model group, positive drug group, low dose of diosgenin group and high dose of diosgenin group at different time points were significantly increased P<0.05), the degrees of toe swelling were significantly increased (P<0.05), and the levels of TNF-α and IL-1β in synovial tissue were significantly increased (P<0.05). Compared with model group, the pain scores of the rats in positive drug group, low dose of diosgenin group and high dose of diosgenin group at different time points were significantly decreased (P<0.05), the degrees of toe swelling were significantly decreased (P<0.05), and the levels of TNF-α and IL-1β in synovial tissue were significantly decreased (P<0.05).Compared with positive drug group, the pain scores of the rats in low dose of diosgenin group and high dose of diosgenin group at different time points were significantly increased (P<0.05), the degrees of toe swelling were significantly increased (P<0.05),and the levels of TNF-α and IL-1β in synovial tissue were significantly increased (P<0.05). Compared with low dose of diosgenin group, the pain score of the rats in high dose of diosgenin group was significantly decreased(P<0.05), the degree of toe swelling was significantly decreased (P<0.05), and the levels of TNF-α and IL-1β in synovial tissue were significantly decreased (P<0.05). The results of HE staining showed that the synovial tissue of the rats in control group was normal, the cells arranged orderly, without edema, hyperemia and proliferation, and had no inflammatory cell infiltration; the synovial tissue of the rats in model group had serious edema and hyperemia,the synovial cells were disordered and proliferaed, and there was a large number of inflammatory cell infiltration; the hyperemia and edema of the synovial tissue of the rats in low dose of diosgenin group were alleviated, and the cell proliferation was slightly reduced; in positive drug group and high dose of diosgenin group, the synovial tissue was slightly edema and congestive, the cells were regularly arranged,and there was small amount of inflammatory cell infiltration and rare proliferation. Compared with control group, the expression levels of TLR2, MyD88 and p-NF-κB p65 proteins in synovial tissue of the rats in model group, positive drug group, low dose of diosgenin group and high dose of diosgenin group were significantly increased (P<0.05);compared with model group, the expression levels of TLR2, MyD88 and p-NF-κB p65 proteins in synovial tissue of the rats in positive drug group, low dose of diosgenin group and high dose of diosgenin group were significantly increased (P<0.05);compared with positive group, the expression levels of TLR2, MyD88 and p-NF-κB p65 proteins in synovial tissue of the rats in low and high doses of diosgenin groups were significantly increased (P<0.05); compared with low dose of diosgenin group, the expression levels of TLR2, MyD88 and p-NF-κB p65 proteins in synovial tissue of the rats in high dose of diosgenin group were significantly decreased(P<0.05).

Conclusion

Diosgenin may improve the symptoms of synovitis rats by inhibiting the TLR2-NF- κB signaling pathway.

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Effect of oxymatrine on proliferation and apoptosis of cervical cancer SiHa cells and its mechanism
Dongli ZHANG,Ruifang FU,Guixia SUN
Journal of Jilin University(Medicine Edition). 2021, 47 (4):  951-957.  DOI: 10.13481/j.1671?587X.20210418
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Objective

To explore the effect of oxymatrine (OMT) on the proliferation and apoptosis of cervical cancer SiHa cells, and to clarify its mechanism.

Methods

The SiHa cells were randomly divided into control group, low dose of OMT group, medium dose of OMT group, and high dose of OMT group. The cells were cultured with RPMI 1640 medium containing dimethyl sulfoxide (DMSO), 0.4, 0.8 and 1.6 g·L-1 OMT. After 24 h of culture, the morphology of cells in various groups was detected by Hoechst33258 staining method; after 24, 48 and 72 h of culture, the proliferation activities of cells in various groups were detected by MTT method; after 48 h, the percentage of cells in different cell cycles and the apoptotic rates in various groups were detected by flow cytometry; RT-qPCR and Western blotting methods were used to detect the expression levels of β-catenin, cyclin D1, B-cell lymphoma-2 (Bcl-2), Bcl-2 assaciated X protein (Bax) mRNA and proteins.

Results

Under fluorescence microscope, the cells in control group grew well, and the blue fluorescence was weak; a small amount of apoptotic bodies were seen in low dose of OMT group; the number of cells in medium dose of OMT group was decreased, and the number of apoptotic bodies were increased; the blue fluorescence in high dose of OMT group was increased, cell disintegration and nuclear shrinkage were found, and the number of apoptotic bodies were increased significantly. Compared with control group, at 24, 48, and 72 h after culture,the proliferation activities of the cells in low dose of OMT group, medium dose of OMTgroup and high dose of OMT group were decreased significantly (P<0.05), the percentage of cells in G0/G1 phase were increased(P<0.05), the percentages of cells in S and G2/M phases were decreased (P<0.05), the apoptotic rates of cells were increased(P<0.05),the expression levels of β-catenin, cyclinD1, Bcl-2 mRNA and proteins were decreased(P<0.05),and the expression levels of Bax mRNA and protein were increased(P<0.05).

Conclusion

OMT can block the SiHa cells in the G1 phase,and inhibit the cell proliferation and induce the apoptosis;its machanism may be related to the inhibition of Wnt/β-catenin signaling pathway.

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Inhibitory effect of diosmetin on myocardial hypertrophy induced by isoproterenol in mice and its mechanism
Pingli WANG,Shenwei ZHANG,Jiang LI
Journal of Jilin University(Medicine Edition). 2021, 47 (4):  958-964.  DOI: 10.13481/j.1671?587X.20210419
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Objective

To investigate the inhibitory effect of diosmetin (Dio) on isoprenaline (Iso)-induced myocardial hypertrophy in the mice, and to elucidate the biological mechanism.

Methods

Twenty-eight mice were randomly divided into control group, model group, low dose of Dio group, and high dose of Dio group. The mice in control group and model group were injected subcutaneously with normal saline or Iso (5 mg·kg-1·d-1) for 14 d, respectively. The mice in low dose and high dose of Dio groups were injected subcutaneously with Iso (5 mg·kg-1·d-1) for 14 d and intraperitoneally with Dio (5 and 20 mg·kg-1·d-1) for 14 d, respectively. After 14 d, the mice were sacrificed, the hearts and tibia were collected, the heart weights and left ventricular weights were weighed, the tibia lengths were measured, and the heart mass index and left ventricular mass index were calculated. The pathomorphology of left ventricular myocardium tissue was observed by HE, FITC-Lectin and Masson staining methods. The expression levels of α-myosin heavy chain (α-MHC) and β-myosin heavy chain (β-MHC) proteins in left ventricular myocardium tissue of the mice were detected by immunohistochemical method. The expression levels of extracellular regulated protein kinase 1/2 (ERK1/2), phosphorylated ERK1/2(p-ERK1/2),c-Jun N-terminal kinase 1/2 (JNK1/2), and phosphorylated JNK1/2(p-JNK1/2) proteins in left ventricular myocardium tissue of the mice were detected by Western blotting method.

Results

Compared with control group, the heart mass index and the left ventricular mass index of the mice in model group were significantly increased (P<0.01), the morphology of left ventricular myocardium tissue was significantly changed, the myocardial fibers were destructed with inflammatory cell infiltration, the cross-areas of myocardial cells were significantly enlarged (P<0.01),the microcirculatory perfusion of myocardium tissue was decreased, and the degree of interstitial fibrosis of myocardium tissue was increased (P<0.01); the expression level of α-MHC protein in the left ventricular myocardium tissue was significantly decreased (P<0.01), and the expression levels of β-MHC, p-ERK1/2 and p-JNK1/2 proteins were significantly increased(P<0.01). Compared with model group,the heart mass index and the left ventricular mass index in different doses of Dio groups were sifnificantly decreased(P<0.05 or P<0.01),the myocardial tissue injury was significantly reduced,the cross-areas of myocardial cells were significantly reduced (P<0.05 or P<0.01), the microcirculatory perfusion of myocardium tissue was enhanced,and the degree of interstitial fibrosis of myocardium tissue was significantly decreased (P<0.05 or P<0.01); the expression levels of α-MHC protein in the left ventricular myocardium tissue were significantly increased (P<0.05 or P<0.01), and the expression levels of β-MHC, p-ERK1/2 and p-JNK1/2 proteins were significantly decreased in a dose-dependent manner(P<0.05 or P<0.01).

Conclusion

Dio can improve Iso-induced myocardial hypertrophy in the mice, and its mechanism may be related to the inhibition of ERK1/2 and JNK1/2 signal pathways.

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Effects of Bozhi glycopeptide on proliferation and adhesion function of human keratinocytes and its mechanism in treatment of psoriasis
Zhenhui MA,shulian LYU,Yaping TIAN
Journal of Jilin University(Medicine Edition). 2021, 47 (4):  965-970.  DOI: 10.13481/j.1671?587X.20210420
Abstract ( 0 )   HTML ( 3 )   PDF (714KB) ( 11 )  
Objective

To investigate the effects of Bozhi glycopeptide on the proliferation of the human keratinocytes (KC) and the expression of β-catenin, and to elucidate the possible mechanism of Bozhi glycopeptide in the treatment of psoriasis.

Methods

In the proliferation function experiment, the HaCaT cells in the logarithmic growth phase were divided into control group and different concentrations (0.011 719, 0.023 438, 0.046 875, 0.093 750, 0.187 500, 0.375 000, 0.750 000 and 1.500 000 mg·L-1) of Bozhi glycopeptide groups. After the cells were treated for 24h, the CCK-8 method was used to detect the proliferation rates of the cells in various groups to determine the effective concentration of Bozhi glycopeptide. The HaCaT cells in the logarithmic growth phase were divided into control group and different concentrations (0.625, 1.250, 2.500, 5.000, 10.000, 20.000, 40.0000 and 80.000 mg·L-1) of Bozhi glycopeptide groups. After the cells were treated for 24 and 48 h, the cell proliferation rates in various groups were detected by CCK-8 method. In the adhesion function experiment, the HaCaT cells in the logarithmic growth phase were divided into control group and different concentrations (10 and 20 mg·L-1) of Bozhi glycopeptide groups. After the cells were treated for 24 and 48 h, the RT-qPCR method was used to detect the expression levels of β-catenin mRNA in the cells in various groups; Western blotting method was used to detect the expression levels of β-catenin protein in the cells in various groups.

Results

In the proliferation function experiment, compared with control group, the cell proliferation rates in 0.011 719-0.093 750 g·L-1 Bozhi glycopeptide groups were significantly reduced after treated for 24 h(P<0.05), which was the effective concentration range of Bozhi glycopeptide. Compared with control group,the proliferation rates of cells in 1.250, 2.500,5.000,10.000,20.000,40.000,and 80.000 mg·L-1 Bozhi glycopeptide groups were significantly reduced after treated for 24 and 48 h(P<0.05 or P<0.01); the proliferation rate was decreased significantly (P<0.01) when the cells were treated with 20 mg·L-1 Bozhi glycopeptide for 48 h. In the adhesion function experiment, compared with control group, the expression levels of β-catenin mRNA in the cells in 10 and 20 mg·L-1 Bozhi glycopeptide groups were decreased significantly after treated for 24 h (P<0.01), and the expression levels of β-catenin mRNA were decreased significantly after treated for 48 h (P<0.01). Compared with control group, after 24 h of treatment, the expression level of β-catenin protein in 10 mg·L-1 Bozhi glycopeptide group was decreased significantly; after 48 h of treatment, the expression levels of β-catenin protein in 10 and 20 mg·L-1 Bozhi glycopeptide groups tended to increase,but there were no significant differences(P>0.05).

Conclusion

Bozhi glycopeptide can inhibit the proliferation of KC at a certain concentration and action time, and can inhibit KC adhesion function by reducing the expression of β-catenin. This may be one of the possible mechanisms of Bozhi glycopeptide in the treatment of psoriasis.

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Effects of targeted silencing of heat shock protein 27 on invasion and migration of oral squamous cell carcinoma CAL27 cells and their mechanisms
Juan CAO,Weibo LI,Xiu GUO,Bo LI,Chunling DONG
Journal of Jilin University(Medicine Edition). 2021, 47 (4):  971-977.  DOI: 10.13481/j.1671?587X.20210421
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Objective

To investigate the effects of heat shock protein 27 (HSP27) on the invasion and migration of oral squamous cell carcinoma (OSCC) CAL27 cells, and to clarify their mechanisms.

Methods

The CAL27 cells were divided into control group [transfected with negative control interference RNA(NC-siRNA)]and experiment group[transfected with HSP27 small interference RNA(HSP-siRNA)].Real-time fluorescence quantitative PCR (RT-qPCR) and Western blotting methods were used to detect the siRNA interference efficiency in the CAL27 cells. Cell scratch assay was used to detect the scratch healing rates of CAL27 cells. Transwell chamber experiment was used to detect the number of invasion and migration CAL27 cells in two groups. RT-qPCR and Western blotting methods were used to detect the mRNA and protein expression levels of epithelial mesenchymal transformation (EMT) marker N-cadherin in the CAL27 cells in two groups.

Results

Compared with control group, the expression levels of HSP27 mRNA and protein in the CLA27 cells in experiment group were significantly decreased (P<0.01).The results of cell scratch experiment showed that compared with control group, the scratch healing rate of the CAL27 cells in experiment group was significantly decreased (P<0.01). The results of Transwell chamber experiment showed that compared with control group, the number of invasion and migration cells in experiment group was significantly reduced (P<0.01). Compared with control group, the expression levels of N-cadherin mRNA and protein in the CAL27 cells in expermient group were significantly reduced (P<0.05).

Conclusion

Targeted silencing of HSP27 can lead to the reduced migration and invasion abilities of the OSCC CAL27 cells, which may be related to the inhibition of EMT.

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Anti-inflammatory effect of honeysuckle extract on acute anterior uveitis mice induced by LPS and its mechanism
Kangning LI,Zhiling SONG,Lilong JIA,Miao CHU,Chaohui XIONG
Journal of Jilin University(Medicine Edition). 2021, 47 (4):  978-983.  DOI: 10.13481/j.1671?587X.20210422
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Objective

To investigate the anti-inflammatory effect of honeysuckle extract on lipopolysaccharide (LPS)-induced acute anterior uveitis (AAU) in the mice, and to illuminate the mechanism.

Methods

Fifty BALB/c mice were randomly divided into control group, model group, and low, medium and high doses of honeysuckle extract groups(n=10).The mice in low, medium and high doses of honeysuckle extract groups were given 250, 500 and 750 mg·kg-1 honeysuckle extract by gavage, and the mice in control group and model group were given normal saline. After administration for 15 d,the mice in other groups except control group were injected with 125 mg·L-1 LPS in the vitreous cavity to establish the AAU mouse models. HE staining was used to observe the pathomorphology of the eye tissue of the mice.Real-time fluorescence quantitative PCR(RT-qPCR) and Western blotting methods were used to detect the expression levels of Toll-like receptor 4(TLR4) and nuclear factor-κB(NF-κB) mRNA and proteins in eye tissue of the mice.ELISA was used to determine the levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-10 (IL-10) in eye tissue of the mice.

Results

The eye tissue of mice in control group was intact and had no inflammatory cell infiltration. Retinal edema occurred and a large number of inflammatory cells infiltrated in tissue of the mice in model group. Compared with model group, the infiltration of inflammatory cells in uvea tissue of the mice in low, medium and high doses of honeysuckle extract groups were decreased to different degrees.Compared with control group, the levels of TLR4 and NF- κB mRNA and proteins in model group were significantly increased (P<0.05), while the levels of TNF-α, IL-6 and IL-10 were significantly increased (P<0.05). Compared with model group,the levels of TLR4 and NF-κB mRNA and proteins in low, medium and high doses of honeysuckle extract groups were significantly decreased (P<0.05), while the levels of TNF-α, IL-6 and IL-10 were significantly decreased (P<0.05). Compared with low dose of honeysuckle extract group,the levels of TLR4 and NF-κB mRNA and proteins in eye tissue of the mice in medium and high doses of honeysuckle extract groups were significantly decreased (P<0.05),while the levels of TNF-α, IL-6 and IL-10 were significantly decreased(P<0.05). Compared with medium dose of honeysuckle extract group,the levels of TLR4 and NF-κB mRNA and proteins in high dose of honeysuckle extract group were significantly decreased (P<0.05), while the levels of TNF-α, IL-6 and IL-10 were significantly decreased(P<0.05).

Conclusion

Honeysuckle extract can down-regulate the expression of inflammatory cells and reduce the inflammatory response of eye tissue in the AUU mice through the TLR4/NF-κB signaling pathway.

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Cyclic fatigue resistance of five kinds of thermally treated nickel-titanium instruments
Qingcui WU,Yingyi WANG,Ang MA,Yiran LI,Shan JIAO,Chengkun WANG,Qiu JIANG
Journal of Jilin University(Medicine Edition). 2021, 47 (4):  984-989.  DOI: 10.13481/j.1671?587X.20210423
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objective

To compare the cyclic fatigue resistance of five kinds of thermally treated nickel-titanium instruments(nickel-titanium files)Reciproc Blue(RB),M-Pro(MP),HyFlex CM(HCM),Orodeka Plex-2.0(OP) and P3-Blue(PB) with the same tip size, and to provide the reference for clinical selection of thermally treated nickel-titanium instruments.

Methods

Twenty new files were selected from RB(25/0.08),MP(25/0.06),HCM(25/0.06),OP(25/0.06)and PB(25/0.06). Each group was further divided into two subgroups with 10 files in each subgroup: RB group, HCM group, MP group, OP group and PB group under 60 ° bending simulated root canal and RB group, HCM group, MP group, OP group and PB group under 90 ° bending simulated root canal.The root canal motor was set according to the speed and torque recommended by the manufacturer.The flexural fatigue resistance of the nickel-titanium files was tested in the curved simulant canals which were made of stainless steel with an inner diameter of 1.5 mm, 60° and 90°angles of curvatures and a radius of curvature of 5 mm.The time from rotation to fatigue fracture (TTF) of each nickel-titanium file was recorded with a timer, and the length of the fracture segment (FL) was measured and recorded with a vernier caliper.After the experiment, the broken nickel-titanium files were collected, and scanning electron microscopic(SEM) evaluation was performed at the fracture sites to investigate the morphology of fracture.

Results

In 60 ° curved simulated root canals, the TTF of nickel-titanium file in RB group was the longest (P<0.05), followed by HCM group and MP group (P< 0.05), and OP group and PB group were the longest(P<0.05);there was no statistically significant difference in TTF between HCM and MP group(P>0.05), and there was no statistically significant difference in TTF between OP group and PB group (P>0.05).In 90 ° curved simulated root canals, the TTF of nickel-titanium file in RB group was the longest(P<0.05), followed by MP group (P<0.05),and HCM group, PB group and OP group were the lowest(P<0.05);there were no significant differences in TTF among HCM group, PB group and OP group (P>0.05).The TTF of nickel-titanium files in each subgroup in simulated root canal bending at 90° were significantly lower than those corresponding subgroups at 60° (P<0.05).There were no statistically significant differences in FL among all groups (P>0.05).The cross sections of nickel-titanium files in each group showed typical characteristics of cyclic fatigue including fatigue striations and fibrous dimples under SEM.

Conclusion

RB nickel-titanium file shows statistically higher cyclic fatigue resistance in artificial canals with 60° and 90° than the other nickel-titanium files.

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Preventive effect of fresh amniotic membrane on intrauterine adhesion and its improvement effect on endometrial receptivity in rabbits
Yanyan FAN,Na WANG,Wen XU,Yajie GE,Lina WANG
Journal of Jilin University(Medicine Edition). 2021, 47 (4):  990-998.  DOI: 10.13481/j.1671?587X.20210424
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Objective

To set up the rabbit intrauterine adhesion (IUA) models, and to explore the effect of placing fresh amniotic membrane in the uterine cavity in the prevention of IUA and its improvement effect on the endometrial receptivity.

Methods

Thirty female New Zealand white rabbits were randomly divided into control group, model group and treatment group, with 10 in each group. Using scraping intrauterine mechanical damage to the endometrium and placing cotton lines soaked in lipid polyaccharides (LPS) in the intrauterine cavity, the rabbit IUA models were established in model group and treatment group. For the rabbits in treatment group, fresh amniotic membrane was placed in the uterine cavity at the same time.The control group was a sham operation group with no uterine treatment. The uterine specimens were collected at the 1st and 2nd weeks. The number of glands and the percentages of endometrial fibrosis areas in endometrium tissue of the rats rabbits in various groups were compared by HE and Masson staining, and the expression levels of integrin αVβ3 and leukemia inhibitory (LIF) proteins in endometrium tissue of the rabbits were detected by immunohistochemical staining.

Results

Compared with control group, the number of glands in endometrium tissue of the rabbits in model group and treatment group were decreased significantly at the lst and 2nd weeks (P<0.05),the percentages of endometrial fibrosis areas were increased significantly(P<0.05),and the expression levels of integrin αVβ3 and LIF proteins in endometial tissue were decreased significantly (P<0.05).Compared with model group, the number of glands in endometrium tissue of the rabbits in treatment group was increased significantly at the 1st and 2nd weeks(P<0.05),the percentages of the endometrial fibrosis areas were decreased significantly (P<0.05), and the expression levels of integrin αVβ3 and LIF proteins in endometrium tissue were increased significantly (P<0.05).In control group and treatment group, compared with the 1st week, there were no statistical differences in the number of glands,the percentages of endometrium fibrosis areas, and the expression levels of integrtin αVβ3 and LIF proteins at 2nd week(P>0.05). But in model group, compared with the 1st week, the percentage of endometrium fibrosis area was decreased significantly at were 2nd week(P<0.01) and the expression levels of integrtin αVβ3 and LIF proteins in endometrium tissue were increased significantly (P<0.05).

Conclusion

The placement of fresh amniotic membrane in the uterine cavity can reduce the degree of IUA and can improve the endometrial receptivity. The effect is fast and lasting.

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Research in clinical medicine
Establishment of human serous ovarian cancer cell culture system in vitro and its application in chemotherapeutic drug sensitivity detection
Jingjing GE,Hongxia XU,Lixia XIE,Kaili DU,Ming SANG,Xiaodong SUN
Journal of Jilin University(Medicine Edition). 2021, 47 (4):  999-1007.  DOI: 10.13481/j.1671?587X.20210425
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Objective

To establish a culture system in vitro of human serous ovarian cancer cells and to theoretical basis to provide the experiment model for the clinical adminstration of the patients with serous ovarian cancer.

Methods

The human serous ovarian cancer cells were isolated and cultured from tumor tissue by type Ⅱ collagenase digestion and differential centrifugation method,and the third of generation serous ovarian cancer cells were used in subsequent experiments. The morphology of ovarian cancer cells was observed under inverted phase contrast microscope, and the cell growth curve was plotted and analyzed. The expression amounts of the human cancer antigen 125 (CA125), paired box gene 8 (PAX8) and epididymis protein 4 (HE4) in the cells, and the expression amounts of embryonic stem cell homologous transcription factor (Nanog) and octamer binding transcription factor 4 (Oct-4) in the cancer stem cells were detected by immunofluorescence. The drug sensitivity test was divided into 5-fluorouracil (5-FU) groups (0, 0.625, 1.250, 2.500, 5.000 mg·L-1), paclitaxel (PTX) groups (0, 0.062 5, 0.125 0, 0.250 0, 0.500 0 mg·L-1), temozolomide (TMZ) groups (0, 0.125, 0.250, 0.500, 1.000 mg·L-1), carboplatin (CBP) groups (0, 0.625, 1.250, 2.500, 5.000 mg·L-1), adriamycin (DOX) groups (0, 0.125, 0.250, 0.500, 1.000 mg·L-1) and melatonin (MLT) groups (0, 0.625, 1.250, 2.500, 5.000 mg·L-1);among them,0 mg·L-1 groups were used as control group; CCK-8 method was used to evaluate the survival rates of the cells.

Results

The primary serous ovarian cancer cells began to adhere to bottom of culture bottles 4 h after enzymatic digestion. The cells grew rapidly and had uniform morphology during continuous passages. The cells grew in a linearly phase during 2-7 d after inoculation, and grow and fuse into pieces after 8-10 days of culture. The results of immunofluorescence assay showed that CA125, PAX8,HE4, Nanog and Oct-4 were highly expressed in the cells. The drug sensitivity test showed that the ovarian cancer cells had high sensitivity to 5-FU,TMZ,CBP and MLT; compared with corresponding control groups,the survival rates of cells in different concentrations of 5-FU groups,TMZ groups and MLT groups and 0.0625 and 0.125 mg·L-1 PTX groups were significantly decreased(P<0.05 or P<0.01).

Conclusion

The culture system in vitro of human serous ovarian cancer cells is successfully established.The cells are sensitive to 5-FU, TMZ, CBP and MLT, which is of guiding significance for the selection of postoperative chemotherapy drugs in the patients with serous ovarian cancer.

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Effects of deoxygenation and reoxygenation speeds on diurnal and nocturnal blood pressure in obstructive sleep apnea patients
Yue CHEN,Lei YU,Xiaoxin LAN,Siqi JIA,Haibo Yuan
Journal of Jilin University(Medicine Edition). 2021, 47 (4):  1008-1013.  DOI: 10.13481/j.1671?587X.20210426
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Objective

To investigate the effect of the speeds of deoxygenation and reoxygenation on the diurnal and nocturnal blood pressure in the obstructive sleep apnea(OSA) patients,and to clarify their mechanisms.

Methods

The information of overnight polysomnography parameters of 106 OSA patients were collected,including sleep parameters and respiratory event parameters,as well as non-invasive multi-point continuous blood pressure monitoring during wakefulness and sleep stages on the monitoring day. The speed of deoxygenation was calculated by the ratio of the latitude value for oxygen desaturation and correspondent duration of desaturation. The reoxygenation speed was calculated similarly. The patients were divided into two groups based on the blood pressure values: OSA complicated with hypertension group (OSA+HT,n=59) and OSA without hypertension group(OSA-HT,n=47). The differences in the speeds of deoxygenation and reoxygenation were analyzed between two groups.The correlations were respectively analyzed between the speeds of deoxygenation,reoxygenation, the diurnal and nocturnal blood pressure and the nocturnal blood pressure fluctuation(NBPF) for all the OSA patients.

Results

The deoxygenation speed and the reoxygenation speed in OSA + HT group was significantly higher than that in OSA-HT group (P<0.01);the deoxygenation speed was positively correlated with the nocturnal systolic blood pressure, diastolic blood pressure, mean arterial pressure, diurnal diastolic blood pressure and mean arterial pressure (r=0.311,P=0.001;r=0.245, P= 0.011;r=0.308, P=0.001;r=0.203,P=0.037;r=0.219,P=0.024), and the reoxygenation speed was positively correlated with the nocturnal systolic blood pressure, diastolic blood pressure, mean arterial pressure,diurnal diastolic blood pressure and mean arterial pressure (r=0.327,P=0.001;r=0.288,P=0.003;r=0.343,P=0.000;r=0.250,P=0.01;r=0.259,P=0.007); there was a positive correlation between both of the deoxygenation speed and the reoxygenation speed and NBPF(r=0.595,P<0.01;r=0.646, P=<0.01).

Conclusion

The damage of deoxygenation and reoxygenation in the OSA patients is directly related to the increase of blood pressure both during wakefulness and sleep stages. The changes of deoxygenation and reoxygenation speeds are the important factors for NBPF in the OSA patients.

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Clinical medicine
Application of three-dimensional reconstruction combined with indocyanine green intraoperative navigation in diagnosis and treatment of liver cancer
Tianyi WANG,Yufeng ZHU,Miao SUN,Wei WANG
Journal of Jilin University(Medicine Edition). 2021, 47 (4):  1014-1021.  DOI: 10.13481/j.1671?587X.20210427
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Objective

To evaluate the application value of three-dimensional reconstruction technology combined with indocyanine green(ICG) fluorescence staining in the diagnosis and treatment of liver tumor, and to explore whether the combined technology can help reduce the incidence of postoperative complications.

Methods

Using retrospective research method, the clinical data of 70 patients with liver cancer were collected; according to whether the patients underwent 3D reconstruction combined with IGF navigation, they were divided into routine preoperative evaluation and hepatectomy group (conventional group) and three-dimensional reconstruction combined with IGF intraoperative navigation group (combined group). The detection and resection of intraoperative lesions, intraoperative blood loss and blood transfusion, operation time, time of postoperative hospital stay, Clavien-Dindo complication classification, and the incidence of major complications after liver resection defined by the International Study Group of Liver Surgery (ISGLS) of the patients were recorded.

Results

In combined group, 15 new lesions were found by intraoperative ICG staining compared with preoperative CT/MRI. The pathological results confirmed that 4 cases were hepatocellular carcinoma, 2 cases were colon cancer liver metastases, and the remaining 9 cases were cirrhotic nodules. The sensitivity was 95.35% and the specificity was 68.75%. Compared with conventional group,the operation time of the patients in combined group was shortened (P<0.05),the Clavie-Dindo complication classification was improved (P<0.05); there were no significant differences in the blood loss,the blood transfusion,the time of postoperative hospital stay, and the incidence of complications defined by ISGLS (P>0.05). The surgical margins of the patients in two groups corresponded the R0 resection standard, and one patient in conventional group presented tumor recurrence at the reexamination 3 months after surgery.

Conclusion

Three-dimensional reconstruction technology combined with ICG staining can improve the safety of the operation and achieve precise positioning through sufficient preoperative evaluation and approach planning, which can effectively shorten the operation time, increase the detection rate of the lesion,has a certain positive impact on reducing the incidence of postoperative complications, and achieves the precise liver resection.

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Bortezomib-induced acute lung injury in patient with multiple myeloma:A case report and literature review
Qiao LI,Zhenzhong SU,Zhaoyong WANG,Yanling XU,Qun ZHANG,Peng GAO,Jie ZHANG
Journal of Jilin University(Medicine Edition). 2021, 47 (4):  1022-1027.  DOI: 10.13481/j.1671?587X.20210428
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Objective

To investigate the pathogenesis, clinical manifestations and treatment strategies of bortezomib-induced acute lung injury in the patient with multiple myeloma,and to provide the reference for the diagnosis and treatment of this disease.

Methods

The clinical data of a patient with acute lung injury caused by bortezomib were collected, and the relevant literatures were summarized and reviewed.

Results

A 48-year-old male patient with multiple myeloma developed fever and dyspnea during the fourth course of VCD regimen (bortezomib, cyclophosphamide and dexamethasone). The patient was diagnosed as bortezomib related acute lung injury by imaging examination, laboratory examination, etiology examination and bronchoscopy. On the premise of discontinuing bortezomib, the patient was given glucocorticoid and noninvasive ventilator assisted ventilation. After treatment, the patient’s condition was improved and the patient was discharged.

Conclusion

Bortezomib treatment-related acute lung injury has low incidence, rapid progress and high mortality, so it is particularly important to identify this kind of disease early. Early discontinuation of bortezomib and glucocorticoid can effectively reduce the mortality and improve the prognosis of suspected the patients.

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Hepatoid adenocarcinoma of stomach:A case report and literature review
Qi ZHU,Lamei LI,Yanjun CAI,Fang XU,Junqi NIU,Wanyu LI
Journal of Jilin University(Medicine Edition). 2021, 47 (4):  1028-1032.  DOI: 10.13481/j.1671?587X.20210429
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Objective

To analyze the clinical features, diagnosis and treatment methods of hepatoid adenocarcinoma of the stomach(HAS), and to improve the understanding of clinicians to this disease.

Methods

The clinical materials of a patient with HAS were collected,and the clinical features, diagnosis and treatment methods were summarized in combination with the relevant literatures.

Results

A 54-year-old man had upper abdominal discomfort for 6 months,and was admitted to the hospital because of gastric cancer. The results of contrast enhanced CT of stomach showed infiltrating ulcerative gastric cancer and multiple liver metastases.Three courses of neoadjuvant chemotherapy were given,after that total gastrectomy was performed,and the pathological results indicated gastric adenocarcinoma.Later, the patient was transferred to our department due to multiple metastatic tumors of the liver.The patient underwent microwave radiofrequency ablation, and the pathological diagnosis was hepatoid adenocarcinoma.The pathology of stomach was confirmed by immunohistochemistry as HAS.After treatment, the survival time was only 9 months.

Conclusion

HAS has no specific clinical features,and can easily be misdiagnosed as gastric adenocarcinoma. The pathological diagnosis and immunohistonchemistry are the golden standard for the diagnosis of HAS. Surgical excision and postoperative adjuvant chemotherapy are the preferred treatment methods.HAS has high malignancy and rapid progression,the patients with HAS should be given early detection and treatment to prolong their survival time.

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Five-year follow-up of oral lichen planus combined with implantation and reparation: A case report and literature review
Yang ZHONG,Degeng XIA,Qingyu ZHANG,Zhulan ZHAO,Yu HUANG,Ning MA,Li ZHANG
Journal of Jilin University(Medicine Edition). 2021, 47 (4):  1033-1037.  DOI: 10.13481/j.1671?587X.20210430
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Objective

To discuss the combined treatment of implant repair of one patient with oral lichen planus(OLP), and to explain the importance of multidisciplinary combined treatment on the restoration of oral function of the patient.

Methods

The clinical data of 5 years of follow-up of one patient with OLP underwent implant repair combined treatment were collected. The related literatures were reviewed to analyze the significance of the application of multidisciplinary combined treatment for the oral function restoration and reconstruction in the patients with OLP.

Results

The clinical manifestion of the patient was that the bilateral moral teeth had become looseness and the patient had a history OLP for 2 years old. Poor oral hygiene, soft gingival texture, dark red, the probing depth (PD) 5-6 mm, attachment loss (AL) 8-9 mm, bleeding on probing (BOP) (+), bilateral posterior teeth loosened Ⅲ°, alveolar bone resorption to 2/3 root were found in the patient; white lesions were seen on the buccal mucosa and the gingival margin of the patient, the mucosa at the lesion was dark red, and erosion and ulcer was seen on the gingival papilla. Regular anti-infective combination therapy (basic periodontal therapy combined with triamcinolone acetonide injection therapy) was performed. When the inflammation remained stable, the Ⅲ° mobile teeth were extracted in turn. Three months after surgery, the implant restoration was performed in the area of tooth loss of the patient. After combined treatment, the condition of OLP of the patient was controlled, the pain and rough discomfort were significantly reduced, the periodontal inflammation was effectively improved. The implant and the surrounding bone tissue were well combined and the chewing function of bilateral posterior teeth was restored.

Conclusion

Multidisciplinary combined treatment can effectively alleviate the clinical symptoms of the patient with OLP and the implant repair of the patient with OLP is realized after the condition is controlled.

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Methodology
Establishment of cultural method of primary SD rat aortic endothelial cells and their identification
Yuanyu TANG,Haiqin LIU,Huagen MA
Journal of Jilin University(Medicine Edition). 2021, 47 (4):  1038-1042.  DOI: 10.13481/j.1671?587X.20210431
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Objective

To establish a simple and efficient cultivating method of the primary SD rat aortic endothelial cells(AECs),and to provide an important carrier and tool cells for the exploration of the molecular biologial characteristics of AECs and the study of cardiovascular and cerebrovascular diseases.

Methods

One SD rat was selected, the chest was opened, and the aorta was removed in sterile condition; the connective tissue and fat outside the blood vessel were removed;the aortic tunica intima was everted and the aorta were cut into 1.0-1.5 mm vascular segments,and the vascular segments were placed in a culture bottle for primary culture. Then the vascular segments were removed 7 d later. The cultured cells were identified by morphological observation and immunocytochemical staining of factor Ⅷ(FⅧ) related antigen.

Results

After 3 d of culture, a small number of cells migrated from the vascular segments to form the island cell mass. From the 10th to 12th days, the adherent cells covered most of the culture bottle surface, and presented a typical “paving stone” mosaic arrangement. The results of immunocytochemical staining of FⅧ showed that the nucleus and the cytoplasm were brownish red,and the expression was positive; the positive cell rate was more than 98%.

Conclusion

Adherent culture method of aortic intima valgus can successfully isolate and culture the primary rat AECs.

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