To investigate the effects of complement 3a （C3a） on the polarization of macrophages in vitro and the effects of complement 3a receptor （C3aR） knockout on the polarization of macrophages in periodontal tissue of mice with chronic periodontitis， the periodontal inflammation and damage degrees of soft and hard tissues， and to clarify the role of C3a-C3aR axis in the occurrence and development of chronic periodontitis and its possible mechanism.

Eighteen 8-week-old female C57BL/6 mice were screened and divided into three groups by genotype： C3ar^+/+ group，C3ar^+/－group and C3ar^－/－ group （n=6）， respectively. The chronic periodontitis models were established by ligating the left maxillary second molars of mice with 4-0 silk thread， and the right maxillary second molars were used as controls. On the 11th day after ligation， the alveolar bone resorption was evaluated； the distance from the cementum enamel junction （CEJ） to the alveolar bone crest （ABC） was detected by Micro-CT；the pathomorphology of the periodontal tissue of the mice in various gorups was observed by HE staining. The levels of inducible nitric oxide synthase （iNOS） and arginase 1 （Arg1） in periodontal tissue of the mice in various groups were detected by immunohistochemical staining. In vitro，the RAW264.7 macrophages were used to establish the inflammatory model by treated with lipopolysaccharide （LPS）；the experiment was divided into blank group， LPS group， C3a group and LPS+C3a group. The polarization of macrophages was observed under light microscope， and the expression levels of iNOS， interleukin-1β （IL-1β） and tumor necrosis factor-α （TNF-α） mRNA were detected by Real-time fluorescence quantitative PCR （RT-qPCR） method.

Compared with C3ar^+/+ and C3ar^+／－ groups， the distance between CEJ and ABC of the mice in C3ar^-/- group was significantly decreased （P<0.01）， inflammatory cell infiltration was decreased； the level of attachment loss was decreased，and the alveolar bone absorption was decreased； the expression level ofiNOS protein was decreased （P<0.05）， whereas the expression level of Arg1 protein was increased （P<0.01）.In the in vitro study， the cells in blank group were round in shape， the cells in C3a group， LPS group and LPS+C3a group were spindled， and the cells in LPS+C3a group showed aggregative growth. The expression levels of iNOS， IL-1β and TNF-α mRNA in LPS group， C3a group and LPS+C3a group were higher than those in blank group （P<0.05）；the expression level IL-1β mRNA in the cells in LPS group and LPS+C3a group were significantly increased（P<0.05）.

In the mouse models of chronic periodontitis， the C3a-C3aR axis may promote the inflammatory response and tissue damage of chronic periodontitis through the polarization of macrophages to M1 type.

To establish a zebrafish xenograft model of human nasopharyngeal carcinoma and explore the inhibitory effect of curcumin on nasopharyngeal carcinoma in vivo and in vitro， and to provide the basis for elucidating the molecular mechanism of anti-nasopharyngeal carcinoma of curcumin.

The nasopharyngeal carcinoma CNE-2 cells were divided into control group and CM-Dil group. The cells were counted for 5 consecutive days， and the fluorescence intensity changes were observed. The zebrafish xenograft model of nasopharyngeal carcinoma was established by microinjection. The zebrafishes were divided into control group， PBS group and model group. The zebrafishes in control group were not treated， the zebrafishes in PBS group were injected with 10 nL PBS buffer， and the zebrafishes in model group were injected with 10 nL cell suspension. The survival number of zebrafishes in each group was counted， and the fluorescence intensity of transplanted tumor of the zebrafishes in model group was observed and quantitatively analyzed by macro stereomicroscope. The morphology of zebrafish transplanted tumors in control group and model group were observed by HE staining. The zebrafishes with the days postfertilization（dpf） was 3 were exposed to feeding water containing different concentrations （0， 0.625， 1.250， 2.500， 5.000，and 7.500， 10.000 μmol·L^－1） of curcumin， the death and deformity were observed；the death rates of zebrafishes were calculated，and the concentration of curcumin in vivo was selected. The zebrafish models were treated with differnent concentrations （0，0.625， 1.250， 2.500， and 5.000 μmol·L^－1） of curcumin for 48 h， the growth and metastasis of the transplanted tumor were detected. The CNE-2 cells were treated with different concentrations（0， 10， and 20 μmol·L^－1） of curcumin for 24 h， the proliferation rates and migration rates were detected by CCK-8 method and wound healing assay.

Compared with control group， there was no significant difference in the number of CNE-2 cells in CM-Dil group （P>0.05）. On the 5th day of the experiment， compared with control group， there was no significant difference in the survival number of zebrafishes in PBS group （P>0.05）； compared with control group and PBS group， the survival number of zebrafishes in model group was decreased significantly （P<0.01）. The red fluorescence intensity in zebrafishes in model group was increased with the prolongation of time. Compared with the first day after injection， the red fluorescence intensity of zebrafishes in model group was increased significantly on the fifth day after injection （P<0.05）.The HE staining results showed that the CNE-2 cells were found in the yolk sac of the zebrafishes in model group， indicating that the models were successfully constructed. The zebrafishes in 0.625， 1.250， 2.500， and 5.000 μmol·L^－1 concentrations of curcumin groups had no death and obvious toxicity. Compared with control group（0 μmol·L^－1 cureumin group），the fluorescence intensities in the zebrafish xenograft tissue in 2.500 and 5.000 μmol·L^－1 cureumin groups were signifricantly decreased（P<0.05 or P<0.01）， and the number of zebrafishes with head and tail metastasis in 5.00 μmol·L^－1 curcumin group was decreased （P<0.05）. The results of CCK-8 method and wound healing assay showed that compared with control group （0 μmol·L^－1 curcumin group），the proliferation rates and migration rates of CNE-2 cells in 10 and 20 μmol·L^－1 curcumin groups were significantly decreased （P<0.05）.

The CNE-2 cells can be successfully implanted in the zebrafishes and to estabish the zebrafish models of nasopharyngeal carcinoma. Curcumin can inhibit the proliferation and metastasis of nasopharyngeal carcinoma in vivo and in vitro， and it has a good prospect of cell anti-tumor.

To explore the anti-fatigue effect of ginseng adventitious root protein （GARP） in the mice and to clarify its mechanism， and provide the experimental basis for the development and utilization of ginseng adventitious root resources.

Forty kunming mice of clean grade were used to establish fatigue animal models by the weight-bearing swimming test. They were randomly divided into control group， low， medium and high doses （0.25， 0.50 and 1.00 g·kg^－1） of GARP groups， and there were 10 mice in each group. The swimming time of the mice in various groups was recorded through the exhaustive swimming experiment. The mice were killed after swimming experiment. The spectrophotometric method was used to detect the blood lactic acid （BLA） and blood urea nitrogen （BUN） levels in the serum of the mice in various groups. The spectrophometric method was used to detect the glutathione （GSH） levels， the superoxide dismutase （SOD） activities and the liver glycogen levels in liver tissue of the mice in various groups， and the spectrophometric method was used to detect the muscle glycogen level in the nuscle tissue of the mice in various groups. The C2C12 myoblasts were divided into control group and different doses （5， 10 and 20 mg·L^－1） of GARP groups.The spectrophotometric method was used to detect the GSH levels， SOD activities， and the glycogen levels in the cells in various groups， and the glucose uptakes of the cells in various groups were determined with phenol sulphuric acid process.The phosphorylation levels of adenosine 5'-monophosphate （AMP）-activated protein kinase （AMPK） （p-AMPK/AMPK） and the expression level of glucose transporter 4 （GLUT4） protein were detected by Western blotting method.

Compared with control group， the swimming time of mice in medium and high doses of GARP groups was increased （P<0.05 or P<0.01）； the BLA and BUN levels in the serum of the mice were reduced （P<0.05 or P<0.01）， the GSH levels and SOD activities in different doses of GAPP groups were increased （P<0.05 or P<0.01）； the liver glycogen and muscle glycogen levels were increased （P<0.05 or P<0.01）. Compared with control group the GSH levels，the SOD activities and the glycogen levels in the C2C12 myoblasts in 5，10 and 20 μg·mL^－1 GARP groups were increased （P<0.05 or P<0.01）， the glucose uptakes were increased （P<0.05 or P<0.01），and the ratios of p-AMPK AMPK and the expression levels of GLUT4 protein were increased （P<0.05 or P<0.01）.

GARP has anti-fatigue effect，and its mechanism may be achieved by activating AMPK/GLUT4 signaling pathway to promote glucose uptake.

To investigate the effect of N-acetylcysteine （NAC） on the apoptosis of mouse MC3T3-E1 preosteoblast cells induced by nicotine， and to clarify its mechanism.

The MC3T3-E1 cells were cultured in vitro and treated with different concentrations （0， 1.0， 2.5， 5.0 and 7.5 mmol·L^－1） of nicotine to induce the MC3T3-E1 cell injury； MTT method was used to determine the proliferation activities of cells in various groups and the half inhibitory concentration（IC₅₀）of nicotine was determined.The MC3T3-E1 cells were divided into control group， nicotine group， NAC group and nicotine+different concentrations（2.5，5.0，and 7.5 mmol·L^－1） NAC groups according the IC₅₀ values， and MTT method was used to determine the proliferation activities of cells in various groups； the optimun concentration of NAC was confirmed.The cells were divided into control group，nicotine （4.25 mmol·L^－1） group，NAC（5.0 mmol·L^－1） group and nicotine（4.25 mmol·L^－1） +NAC（5.0 mmol·L^－1） group.The levels of mitochondrial ROS （mitoROS） was assessed with MitoSOX fluorescence staining method； Flow cytometry was used to detect the apoptic rates of MC3T3-E1 cells in various groups； Western blotting method was used to detect the expression level of apoptosis-related proteins B-cell lymphoma-2 （Bcl-2） and Bcl-2 associated X proteins （Bax）in the cells in various groups， and the ratios of Bax/Bcl-2 was calculated.

Compared with 0 mmol·L^－1 nicotine group， the proliferation activities of MC3T3-E1 cells in different concentrations of nicotine groups were significantly decreased （P<0.05） in a dose-dependent manner. The IC₅₀ of nicotine was （4.25±0.03） mmol·L^－1.Compared with control group，the proliferation activity of MC3T3-E1 cells in 4.25 mmol·L^－1 nicotine group was significantly decreased （P<0.05）； compared with nicotine group， the proliferation activites of MC3T3-E1 cells in nicotine+different concentrations of NAC groups were increased（P<0.05）；the proliferation activity of cells in nicotine+5.0 mmol·L^－1 NAC group was the highest. Compared with control group， the levels of mitoROS in the cells in nicotine group was significantly increased （P<0.05）， the apoptotic rate of cells was significantly increased （P<0.05），and the ratio of Bax/Bcl-2 was increased （P<0.05）；compared with nicotine group， the levels of mitoROS in the cells in NAC and nicotine+NAC group were significantly decreased （P<0.05）， the apoptotic rate was significantly reduced （P<0.05），and the ratio of Bax/Bcl-2 in nicotine+NAC group was reduced （P<0.05）.

NAC can alleviate the apoptosis of MC3T3-E1 cells induced by nicotine， and its mechanism may be related to the mitochondrial oxidative stress.

To investigate the expression of miR-150-5p in kidney tissue of the diabetic nephropathy （DN） model mice and the effect of miR-150-5p on the MPC5 mouse podocyte injury induced by high glucose， and to elucidate its mechanism.

Twenty Kunming mice were randomly divided into control group （n=8， normal feeding） and DN group （n=12， high fat diet feeding combined with intraperitoneal injection of streptozotocin to establish the DN models）.The expression levels of miR-150-5p in kidney tissue of the mice in two groups were detected by real-time fluorescence quantitative PCR （RT-qPCR）， and the expressions of brain acid soluble protein 1 （BASP1） in kidney tissue of the mice in two groups were observed by immunohistochemical and immunofluorescence methods.The MPC5 cells were cultured in medium without interferon-γ（IFN-γ） for 2 weeks to differentiate and mature， then the cells were divided into control group and high glucose treatment （HG） group. The cells in HG group were treated with 30 mmol·L^－1 D-glucose for 12， 24， 48 and 72 h， respectively. The expression levels of miR-150-5p and BASP1 mRNA and protein in the MPC5 cells were detected by RT-qPCR and Western blotting methods， respectively. Bioinformatics and luciferase reporter assays were used to evaluate whether miR-150-5p had a targeting relationship with BASP1.The miR-NC， miR-150-5p mimic， miR-150-5p mimic+pc-DNA3.1-SC and miR-150-5p mimic+pc-DNA3.1-BASP1 were transformed into the mature MPC5 cells， respectively， and they were divided into normal condition+miR-NC （Con+NC） group， HG condition+miR-NC（HG+NC） group， HG condition+miR-150-5p mimic （HG+mimic）group，and HG condition +miR-150-5p mimic+pc-DNA3.1-SC（HG+ mimic+SC） group and HG condition +miR-150-5p mimic+pc-DNA3.1-BASP1 （HG+ mimic+BASP1） group. The viabilities of MPC5 cells in various groups were detected by CCK-8 assay； the reactive oxygen species （ROS） levels and lactate dehydrogenase （LDH） activities of MPC5 cells in various groups were detected by ELISA method， and the apoptotic rates of MPC5 cells in various groups were detected by flow cytometry.

Compared with control group， the expression level of miR-150-5p in kidney tissue of the mice in DN group was decreased （P<0.01）， while the expression amount of BASP1 protein was increased， and BASP1 was co-localized with the podocytes. Compared with control group， the expression levels of BASP1 protein in the MPC5 cells in HG group were increased （P<0.01），and the expression levels of miR-150-5p were decreased （P<0.05 or P<0.01） in a time dependent manner. BASP1 was a direct target of miR-150-5p. Compared with Con+NC group， the viability of MPC5 cells in HG+NC group was significantly decreased （P<0.01）， while the ROS level， LDH activity and apoptotic rate were significantly increased （P<0.01）. Compared with HG+NC group， the viabilities of MPC5 cells in HG+mimic and HG+mimic+SC groups were significantly increased （P<0.01）， while the ROS levels， LDH activities and apoptotic rates were significantly decreased （P<0.01）.Compared with HG+mimic+SC group， the viability of MPC5 cells in HG+mimic+BASP1 group was significantly decreased （P<0.01）， while the ROS level， LDH activity and apoptotic rate were significantly increased （P<0.01）.

The decreased expression of miR-150-5p may be one of the mechanisms of podocyte injury in the DN models. Over-expression of miR-150-5p may inhibit the HG-induced podocyte injury by targeting BASP1.

To observe the effects of Schisandrin B（SchB）， one of the monomers of lignans from Schisandra chinensis， on the proliferation， cell cycle， type Ⅰ collagen and type Ⅲ collagen expression levels in the neonatal SD rat cardiac fibroblasts（CFb） induced by angiotensinⅡ（AngⅡ）， and to explore the mechanisms of phosphatidylinositol 3-kinase/protein kinase B/cell cycle inhibiting protein（P27kip1）（PI3K/Akt/P27kip1）signal pathways.

The myocadium tissue was obtained from the neonatal 1－3 d of SD rats.The CFb were cultured by trypsin digestion and differential adhesion method and identified as the required CFb.The experiment was divided into control group， model group， LY294002（PI3K inhibitor） group，SchB group，LY294002+SchB group.The CFb were stimulated with 0.1 μmol·L^－1 AngⅡ，and the inhibitory rates of proliferation of CFb were detected by MTT colorimetry after treated for 24 h.The expression levels of typeⅠand type Ⅲ collagen proteins were detected by immunohistochemistry method，the percentages of CFb in different cell cycle in various groups were detected by flow cytometry.The expression levels of PI3K， Akt， phosphorylated Akt（p-Akt） and P27kip1 proteins in the CFb in various groups were detected by Western blotting method.

Compared with control group， the inhibitory rate of proliferation of the CFb in model group was increased（P<0.01）， the exspression levels of typeⅠ collagen and type Ⅲ cellagen proteins were increased（P<0.01），the percentage of CFb in G₀/G₁ phase was decreased（P<0.01）， the percentage of CFb in S phase and the proliferation index（PI） were increased（P<0.01），the expression levels of PI3K and Akt proteins in the CFb were increased（P<0.01）， and the expression level of P27kip1 protein was decreased（P<0.01）；compared with model group， the inhibitory rates of prolifenition of the CFb in LY294002， SchB，and LY294002+SchB groups were significantly decreased（P<0.05 or P<0.01）， the expression levels of typeⅠand type Ⅲ collagen proteins were significantly decreased（P<0.05 or P<0.01）， the percentages of CFb in G₀/G₁ phase were increased（P<0.05 or P<0.01）， the percentages of CFb in S phase and PI were decreased（P<0.05 or P<0.01）， the expression levels of PI3K and p-Akt proteins were decreased（P<0.05 or P<0.01）， and the expression levels of p27kip1 proteins were increased（P<0.05 or P<0.01）.

SchB can inhibit the AngⅡ-induced CFb proliferation and collagen deposition， and its mechanism may be related to inhibiting the PI3K/Akt/P27kip1 signal transduction pathways.

To compare the degrees of occurrence of microleakage of class Ⅴ cavities of the isolated teeth filled with three flowing materials Z350XT flowing resin， Beautifil Flow Plus F00 and PRG Barrier Coat and the influence of saliva， and to select the suitable materials for wedge-shape defects.

A total of 42 isolated first premolars were selected，and 24 isolated teeth were used in the experiment without saliva and were randomly divided into 4 groups： Z350XT flowing resin， Beautifil Flow Plus F00， PRG Barrier Coat and glass ionomer cement （GIC） （n=6）. A total of 18 isolated first premolars were used in the experiment with saliva and were randomly divided into 3 groups： Z350XT flowing resin， Beautifil Flow Plus F00 and PRG Barrier Coat （n=6）.The class Ⅴ cavity was made on the facial surface of each isolated tooth in the experiment without saliva and was filled with the corespondent materials. The samples in the experiment with saliva were soaked in artificial saliva after coated with 3M single bond universal adhesive， and then were filled with the correspondent materials. After thermocycling， the apical foramens were closed with plastomer and the samples were stained with methylene blue solution. The sections were made. The dye depths，namely microleakage degrees，of the isolated tooth in each group were observed and measured under stereoscopic microscope，and the microleakage degree scores were calculated.

In the experiment without saliva，the microleakage degree and the microleakage score in GIC group were higher than those of the other groups without saliva（P<0.05）；the microleakage degree and the microleakage degree score in PRG Barrier Coat group were higher than those in 350XT flowing resin group and Beautifil Flow Plus F00 group（P<0.05）；the microleakage degrees and the microleakage degree scores in gingival walls in various groups were higher than those of occlusal walls of the same material （P<0.05）.In the experiment with saliva，there were no differences in the microleakage eakage degrees and the microleakage degree scores among various groups（P>0.05）；the microleakage degrees and the microleakage degree scores in each group with saliva were higher than those of the same material without saliva （P<0.05）.

As materials for filling class Ⅴ cavities， Z350XT flowing resin and Beautifil Flow Plus F00 are superior to PRG Barrier Coat and GIC. It should be widely promoted in clinical practice. Prevention of saliva contamination is critical to reducing microleakage.

To study the antioxidant activity in vitro of Bupleurum chinense polysaccharides （BCP），and to clarify its protective effect on D-galactose （D-gal）-induced proximal convolve tubule HK-2 cells of human kidney.

BCP was extracted by hot water extraction method. The antioxidant activities of BCP including the hydroxyl radical scavenging rate，the total antioxidant capacity and the anti-superoxide anion activities were detected，and compared to the antioxidant activities of vitamin E.The HK-2 cells were treated with different concentrations of BCP and D-gal，respectively，and the proliferation rates of HK-2 cells were detected；the optimum concentrations of BCP and D-gal were confinmed.D-gal was used to establish the oxidative damage model of proximal convolve tubule of human kidney.The HK-2 cells were divided into control group，D-gal group，and 25，50，100，200，400 mg·L^－1 BCP+D-gal groups；MTT method was used to detect the proliferation rates.The HK-2 cells were divided into control group，D-gal group，BCP+D-gal group and vitamin E+D-gal group；senescence-associated β-galactosidase（SA-β-Gal） staining was used observe the SA-β-gal staining of the HK-2 cells in various groups and the percentages of senescent the HK-2 groups cells were calculated.

The antioxidant activity detection results showed that compared with vitamin E group with the same concentration， the scavenging rates of hydroxyl free radical in different concentrations of BCP groups were increased（P<0.05）， and the total antioxidant capacitie were increased（P<0.05）； the anti-superoxide anion activities in 5，10，and 15 g·L^－1 BCP groups were significantly increased（P<0.05）， and the antioxidant levels in vitro in different concentrations of BCP groups were higher than those in vitamin E groups with the same concentration. Compared with control group，the proliferation rates of HK-2 cels in 25－400 mg·L^－１ BCP groups were increased in a concentration-dependent manner after treated for 24 h， and the proliferation rate of HK-2 cells in 200 mg·L^－1 BCP group was the highest （P<0.05）. Compared with control group， the proliferation rates of HK-2 cells in different concentrations of D-gal groups were decreased in a time-dose dependent manner； when the treatment conditions were 20 g·L^－1 and 48 h， the proliferation rate of cells was decreased to 64.77% of that in control group （P<0.05）.Compared with control group，the proliferation rate of HK-2 cells in D-gal group was significantly decreased（P<0.05）； compared with D-gal group， the proliferation rates of HK-2 cell in 100－400 mg·L^－1 BCP group increased in a dose-dependent manner （P<0.05 or P<0.01）. Compared with control group， the number of SA-β-gal positive staining cells in D-gal group was significantly increased（P<0.05）， and the percentage of senescent cells was increased （P<0.05）； compared with D-gal group， the number of SA-β-gal positive staining cells in BCP+D-gal group and vitamin E+D-gal group was significantly reduced（P<0.05）， and the percentages of senescent cells were decreased （P<0.05）.

BCP has a strong antioxidant activity in vitro and protects the oxidative damage of HK-2 cells induced by D-gal by inhibiting cell senescence.

To explore the effect of overexpression of B-cell lymphoma-2（Bcl-2） associated X protein （Bax） inhibitor 1 （BI-1） on the cardiomyocyte apoptosis in the rats with acute myocardial infarction （AMI）， and to clarify its mechanism.

A total of 60 rats were randomly divided into sham operation group， model group， empty adenovirus （Ad-NC） group and BI-1 adenovirus （Ad-BI-1） group； there were 15 rats in each group. Except for sham group， the rats in other groups were ligated in the left anterior descending coronary arteries to establish the AMI rat models. Meanwhile， the rats in Ad-NC group and Ad-BI-1 group were injected with adenovirus-packaged empty plasmid and BI-1 overexpression plasmid in the myocardial infarction area， respectively. The cardiac function parameters left ventricular ejection fraction （LVEF）， left ventricular fraction shortening （LVFS）， left ventricular end diastolic diameter （LVEDD） and left ventricular end systolic diameter （LVESD） values of the rats were detected 72 h after operation； TTC staining method was used to detect the percentages of myocardial infarction areas of the rats in various groups； TUNEL method was used to detect the apoptotic rates of cardiomyocytes of the rats in various groups； colorimetric method was used to detect the activities of Caspase-3 in myocardium tissue of the rats in various groups； RT-qPCR method was used to detect the expression levels of BI-1 mRNA of the rats in various groups； Western blotting method was used to detect the expression levels of BI-1， Bcl-2， Bax， and endoplasmic reticulum stress related proteins GRP78， IRE1， p-IRE1，JNK，p-JNK proteins in myocardium tissue of the rats in various groups；the ratios of the p-IRE1/IRE1 and p-JNK/JNK were calculated.

Compared with sham operation group， the LVEF and LVFS of the rats in model group were significantly reduced （P<0.05），while the LVEDD and LVESD，the percentage of myocardial infarction area， the Caspase-3 activity in myocardium tissue， and the apoptodtic rates of cardiomocytes were increased （P<0.05）， the expression levels of BI-1 mRNA and protein， and the expression levels of Bcl-2 protein in myocardium tissue of the rats in model group were significantly reduced （P<0.05）， and the expression levels of Bax， and GRP78 and the ratios of p-IRE1/IRE1 and p-JNK/JNK were significantly increased （P<0.05）.Compared with model group， the LVEF and LVFS of the rats in Ad-BI-1 group were significantly increased （P<0.05）， while the LVEDD and LVESD were markedly decreased（P<0.05）； the percentage of myocardial infarction area，the apoptotic rate of cardiomyocytes，and the Caspase-3 activity in myocardium tissue were decreased （P<0.05）； the expression levels of BI-1 mRNA and protein and the expression level of Bcl-2 protein in myocardium tissue of the rats were significantly increased （P<0.05），and the expression levels of Bax and GRP78 and the ratios of p-IRE1/IRE1 and p-JNK/JNK were significantly decreased （P<0.05）； the changes of the above indicators in Ad-NC group had no siginficant differences（P>0.05）.

Overexpression of BI-1 reduces the level of cardiomyocyte apoptosis by inhibiting the endoplasmic reticulum stress pathway， thereby improving the cardiac function of the AMI rats.

A total of 40 SPF grade SD rats were randomly divided into control group，DR model group，15 mg·kg^－1 paeonol group and 30 mg·kg^－1 paeonol group；there were 10 rats in each group.Except control group，the rats in other groups were intraperitoneally injected with 55 mg·kg^－1 streptozotocin（STZ） to establish the DR medels.After successeful modeling，the rats in control group and DR model group were subcutaneously injected with normal saline，and the rats in 15 and 30 mg·kg^－1 paeonol groups were subcutaneously injected with 15 and 30 mg·kg^－1 paeonol sodium sulfonate.The rat retina tissue was harvested for real-time fluorescence quantitative PCR （RT-qPCR）， Western blotting， and ELISA assays and HE staining was performed to observe the pathomorphology of retina tissue of the rats in various groups.The human retinal microvascular endothelial cells （HRMECs） were cultured in high glucose （HG，33 mmol·L^-1 glucose medium）. Different doses （0， 5， 25， 50， 70 and 100 mg·L^-1） of paeonol were used to intervene the HRMECs and the HRMECs treated with HG for 24 h， respectively；the cell viabilities and apoptotic rates of cells in various groups were detected by CCK-8 assay and flow cytometry， respectively.The optimum dose used in the follow-up experiment was confirmed.The transfected HRMECs were divided into control group， HG group， HG+NC inhibitor group， HG+miR-802-5p inhibitor group， HG + paeonol （25 mg·L^－1） group，HG+paeonol +NC mimic group， and HG+ paeonol + miR-802-5p mimic group.The expression levels of miR-802-5p in the retina tissue and the cells in various groups were determined by RT-qPCR method，and the association between miR-802-5p and SIRT6 in the HRMECs was determined by luciferase reporter gene assay and RNA co-immunoprecipitation assay；the protein expression levels of silent information regulator 6 （SIRT6）， pigment epithelium derived factor （PEDF） and vascular endothelial growth factor （VEGF） in the retina tissue of the rats and the cells in various groups were determined by Western blotting method. The levels of reactive oxygen species （ROS） and the activities of catalase （CAT） and superoxide dismutase （SOD） in the cells in various groups were measured by ELISA.

Compared with control group， the expression levels of miR-802-5p and VEGF protein in the retina tissue of the rats in DR model group were significantly increased （P<0.01）， and the protein expression levels of SIRT6 and PEDF were decreased （P<0.05）， the ROS level was increased （P<0.01）， and the activities of CAT and SOD were decreased （P<0.05）.Compared with DR model group，the expression levels of miR-802-5p and the expression levels of VEGF protein in retinal tissue of the rats in different dose of paeonal groups were significantly decreased（P<0.05），the expression levels of SIRT6 and PEDF were signifricantly increased（P<0.01），the ROS levels were significantly decreased （P<0.05 or P<0.01），and the activities of CAT and SOD were signifricantly increased（P<0.05 or P<0.01）.The HE staining results showed that the retinal structure of the rats in control group was clear， the cells in the inner and outer nuclear layers were arranged regularly， and no neovascularization was observed； in DR model group， the retinal structure was unclear， the nerve fiber layer was edematous， the inner and outer nuclear layer cells were arranged loosely， and neovascularization was observed； the retinal structures were clear and the cells in the inner and outer nuclear layers were arranged in an orderly manner in different doses of paeonol groups， and the neoangiogenesis was significantly improved compared with that in model group. Compared with control group，the apoptotic rate of cells in HG group was increased（P<0.01）；compared with HG group，the apoptotic rate of cells in HG+paeonol group was decreased（P<0.01）.Compared with control group， the expression level of miR-802-5p in the cells in HG group was significantly increased （P<0.01）；compared with HG+NC inhibitor group，the expression level of miR-802-5p in the cells in HG+miR-802-5p inhibitor group was significantly decreased（P<0.01）.The results of luciferase reporter gene assay and RNA co-immunoprecipitation assay showed that SIRT6 was a target gene of miR-802-5p.Compared with control group，the expression levels of SIRT6 and PEDF proteins and the activities of CAT and SOD in the cells in HG group were significantly decreased（P<0.01），and the expression level of VEGF protein and the ROS level were significantly increased（P<0.01）；compared with HG group， the expression level of VEGF protein in the cells in HG+miR-802-5p inhibitor group was decreased （P<0.01），the expression levels of SIRT6 and PEDF proteins were increased （P<0.05），the ROS level was decreased （P<0.05）， and the activities of CAT and SOD were increased significantly （P<0.01）.Compared with HG group，the ROS level in the cells in HG+paeonol group was decreased（P<0.01），and the activities of CAT and SOD were increased（P<0.01）；compared with HG + paeonol + NC mimic group， the ROS level in the cells in HG + paeonol + miR-802-5p mimic group was increased （P<0.05）， and the activities of CAT and SOD were significantly decreased （P<0.01）.

Paeonol may improve the DR progression through the miR-802-5p/SIRT6 axis.

To construct the overexpression vector for the mouse Nuclear receptor subfamily 1 group D member 1（NR1D1） gene， and to further analyze the basic properties of NR1D1 protein.

The total RNA was extracted from the mouse liver tissue， and the cDNA was obtained by reverse transcription. PCR was utilized to amplify the coding fragment for the NR1D1 gene， and the NR1D1 CDS fragment was then ligated into the pcDNA3.1 vector by homologous recombination reactions. The recombinant plasmids were identified by restriction enzyme digestion and sequencing， and the recombinant plasmid was named as pcDNA3.1-NR1D1， and then the pcDNA3.1 plasmids and pcDNA3.1-NR1D1 recombinant plasmids were transfected respectively into the HEK293T cells and named as control group and pcDNA3.1-NR1D1 transfection group.After 48 h，the extraction of total RNA and total protein from cell samples were performed，and the expression levels of NR1D1 mRNA and protein were detected by real-time fluorescence quantitative PCR（RT-qPCR） and Western blotting methods， respectively. Meanwhile， the similarities of NR1D1 coding sequence（CDS） between the mice and other species were analyzed by DNAStar software and MEGA7 software， and the phylogenetic trees were constructed. In addition， the amino acid composition，hydrophilicity/hydrophobicity， transmembrane regions， signal peptides， secondary and tertiary structures of the NR1D1 protein were analyzed with online tools， such as ExPASy， ProtScale， SingalP5.0， TMHMM-2.0， PhosphoSitePlus^?， SOPMA and SWISS-MODEL.

The restriction enzyme digestion and sequencing results showed that the overexpression vector pcDNA3.1-NR1D1 was successfully constructed. Compared with control group， the expression levels of NR1D1 mRNA and protein in the cells in pcDNA3.1-NR1D1 transfection group were significantly increased （P<0.01）. In addition， the bioinformatics analysis revealed that the similarities of the Mus musculus NR1D1 CDS region with Homo sapiens， Rattus norvegicus， Cricetulus griseus， Pan troglodytes， Oryctolagus cuniculus， Canis lupus familiaris， Sus scrofa， Equus caballus， Bos taurus， Ovis aries， Capra hircus， Felis catus and Danio rerio were 90.0%， 94.8%， 86.5%， 90%， 89.6%， 88.3%， 89.7%， 89.8%， 88.8%， 89.1%， 89.1% and 65.1%， respectively. The phylogenetic tree showed that the Mus musculus NR1D1 gene was the closest genetically to the Cricetulus griseus and Rattus norvegicus， and the most distant from Danio rerio. Mouse NR1D1 protein was a hydrophobic basic protein， not a secreted protein or a transmembrane protein， which contained 12 phosphorylation sites and 10 ubiquitination sites. The secondary structure of mouse NR1D1 protein included 54.63% random coil， 26.50% α-helix， 12.68% extended chain， and 6.18% β-turn， and NR1D1 protein had larger similarity and less variation in tertiary structure compared with the human.

The mouse NR1D1 gene overexpression vector pcDNA3.1-NR1D1 is successfully constructed， and its overexpression efficiency is verified in the HEK293T cells， which provide the basis for further research on the function of NR1D1 gene and protein.

To investigate the regulatory effect of miRNA-27a on the immune function of the experimental pulmonary tuberculosis model rats， and to clarify its possible mechanism.

The rat models of pulmonary tuberculosis were established by injecting Mycobacterium tuberculosis suspension through tail vein， after successful modeling， the model rats were randomly divided into model group， miR-27a agomir control （agomir-NC） group and miR-27a agomir （agomir） group，and another normal rats were used as control group； there were 12 mice in each groups.The rats in agomir-NC group or miR-27a agomir group were injected with agomir-NC or miR-27a agomir via tail vein once a day for 5 consecutive days. Three weeks later，the spleen and thymus indexes of the rats in various groups were calculated， the pathomorphology of lung tissue of the rats in various groups were observed by HE staining； the percentages of T lymphocyte subsets in peripheral blood of the rats in various groups were detected by flow cytometry； the levels of interleukin-1β （IL-1β）， interleukin-6 （IL-6） and tumor necrosis factor-α （TNF-α） in lung tissue of the rats in various groups were determined by enzyme linked immunosorbent assay （ELISA）； the expression levels of miR-27a in lung tissue of the rats in various groups were detected by real-time fluorescence quantificative PCR （RT-qPCR） method； the expression levels of Toll-like receptor 4 （TLR4）， nuclear factor kappa-B （NF-κB） inhibitor α （IκBα）， phosporylated-IκB（p-IκB） and NF-κB p65 in lung tissue were detected by Western blotting method.

Compared with control group，the lung injury of rats in model group was serious， the spleen and thymus indexes， the percentages of CD4+T cells in peripheral blood， the expression level of miR-27a and the expression level of IκB protein in lung tissue were significantly decreased （P<0.05）； the percentages of CD8+T cells in peripheral blood， the levels of IL-1β， IL-6， TNF-α and the expression levels of TLR4， p-IκB and NF-κB p65 proteins in lung tissue were significantly increased （P<0.05）. Compared with model group， the pathomorphology of lung tissue of the rats in agomir group was significantly improved， the spleen and thymus indexes，the percentages of CD4+T cells in peripheral blood， the expression level of miR-27a and the expression level of IκB protein in lung tissue were significantly increased （P<0.05）； the percentage of CD8+T cells in peripheral blood， the levels of IL-1β， IL-6， TNF-α and the expression levels of TLR4， p-IκB and NF-κB p65 proteins in lung tissue of the rats were significantly decreased （P<0.05）； there were no statistically significant differences in the above indexes of the rats in agomir-NC group （P>0.05）.

Overexpression of miR-27a may improve the lung injury in the pulmonary tuberculosis rats by inhibiting activation of TLR4/NF-κB signaling pathway， reducing the release of inflammatory factors and regulating the body's immune response.

To investigate the effects of salvianolate （Sal） on the osteoclast differentiation and bone resorption in the osteoporotic rats， and to clarify its mechanism.

Castration was used to establish the rat models of osteoporosis. A total of 30 rats were randomly divided into sham operation group， model group and Sal group （given with 40 mg·kg^－1 Sal by intraperitoneal injection once two days 4 weeks after modeling for 4 weeks）.HE staining method was used to observe the pathomorphology of femur tissue of the rats in various groups； dual-energy X-ray bone densitometer was used to detect the femur bone mineral density （BMD） of the rats in various groups； three-point bending test was used to detect the maximum loads of femur of the rats in various groups. In in vitro experiment， 30 μg·L^－1 macrophage colony stimulating factor （M-CSF） and 100 μg·L^－1 nuclear factor kappa B （NF-κB） ligand （RANKL） were used to induce the differentiation of bone marrow macrophages （BMMs）. The cells were divided into control group， RANKL group， RANKL +Sal group （10 mg·L^－1 Sal）， RANKL +Sal +vector group （10 mg·L^－1 Sal and 2 mg·L^－1 empty vector）， RANKL +Sal +LV-FKBP1A group ［10 mg·L^－1 Sal and 2 mg·L^－1 lentivirus-mediated FKBP1A overexpression vector （LV-FKBP1A）］， RANKL +Sal+LV-FKBP1A+QNZ group （10 mg·L^－1 Sal， 2 mg·L^－1LV-FKBP1A and 10 nmol·L^－1 NF-κB inhibitor QNZ）. MTT assay was used to detect the proliferation activities of the cells in various groups； flow cytometry was used to detect the apoptotic rates of cells in various groups； Co-IP experiment was used to verify the interaction between SMAD2 and FKBP1A； Western blotting method was used to detect the expression levels of SMAD family member 2 （SMAD2）， FK506-binding protein 1A （FKBP1A） and phosphorylated NF-κB P65 （p-P65） proteins in rat bone tissue and cells in various groups. Enzyme-linked immunosorbent assay （ELISA） was used to detect the levels of tartrate-resistant acid phosphatase 5b （TRAP5b） in serum of the rats and cells in various groups；real-time fluorescence quantitative PCR （RT-qPCR） was used to detect the expression levels of cathepsin K （CTSK） and calcitonin receptor （CTR） mRNA in bone tissue of the rats and cells in various groups.

The in vivo experimental results showed that compared with model group， the number of bone trabeculae in Sal group， the BMD and the maximum load of femur of the rats were significantly increased （P<0.01）， and the level of TRAP5b in serum was decreased（P<0.01）， the expression levels of SMAD2 and FKBP1A proteins and CTSK and CTR mRNA in bone tissue were significantly reduced （P<0.01）. The Co-IP experiment results confirmed that SMAD2 interacted with FKBP1A protein. The in vitro experimental results showed that compared with RANKL group， the proliferation activity of cells in RANKL+Sal group was significantly decreased（P<0.01），the apoptotic rate was significantly increased（P<0.01），and the TRAP5b level and the expression levels of SMAD2， FKBP1A and p-P65 proteins and CTSK and CTR mRNA were significantly reduced （P<0.01）.Compared with RANKL+ Sal +vector group，the apoptotic rate in RANKL+Sal+LV-FKBP1A group was significantly reduced（P<0.01），and the TRAP5b level and the expression levels of SMAD2， FKBP1A and p-P65 proteins and CTSK and CTR mRNA in the cells were significantly increased （P<0.01）.Compared with RANKL+ Sal+LV-FKBP1A group，the apoptotic rate in RANKL +Sal +LV-FKBP1A +QNZ group was significantly increased （P<0.01），and the TRAP5b level and the expression levels of SMAD2， FKBP1A and p-P65 proteins and CTSK and CTR mRNA in the cells were significantly reduced （P<0.01）.

Sal inhibits the activation of NF-κB by regulating the interaction between SMAD2 and FKBP1A protein， and alleviates the osteoporosis in the rats.

To investigate the effects of betulinic acid （BEA） on the proliferation， apoptosis， migration and invasion of gastric cancer MGC-803 cells， and to elucidate their mechanisms.

The MGC-803 cells were divided into control group and different doses of BEA groups，and the cells were cultured with DMEH high sugar medium containing 0，2.5，5.0，10.0，20.0，40.0，and 80.0 μmol·L^－1 BEA. The proliferation rates， apoptotic rates， migration rates and the number of invasion MGC-803 cells were detected by CCK-8 method， flow cytometry， scratch test and Transwell method， respectively. The expression levels of SMAD homology 7（SMAD7），transforming growth factor-β receptor 1（TGF-βR1）， phosphorylated SMAD2（p-SMAD2），phosphorylated SMAD3（p-SMAD3），SRY-box transcription factor 4（SOX4）， zinc finger E-box- binding protein 2（ZEB2）， matrix metalloproteinase-9（MMP-9）， Snail and Slug proteins in the MGC-803 cells in various groups were detected by Western blotting method.

Compared with control group，the proliferation rates of cells in 2.5， 5.0， 10.0， 20.0， 40.0 and 80.0 μmol·L^－1 BEA groups were decreased after 24， 48 and 72 h of incubation（P<0.05）.After 72 h of incubation， the apoptotic rates in 2.5， 5.0， 10.0 and 20.0 μmol·L^－1 BEA groups were increased compared with control group （P<0.05）. After 24 h of incubation， compared with control group， the migration rates in 2.5， 5.0， 10.0 and 20.0 μmol·L^－1 BEA groups were decreased （P<0.05），and the number of invasion cells was decreased（P<0.05）. The results of Western blotting method revealed that compared with control group，the expression level of SMAD7 protein in 20 μmol·L^－1 BEA group was increased（P<0.05），and the expression levels of TGF-βRI， p-SMAD2， p-SMAD3， SOX4， ZEB2， MMP-9， Snail and Slug proteins were decreased after 48 h of incubation.

BEA regulates the proliferation， apoptosis， migration and invasion of gastric cancer cells by up-regulating the expression of SMAD7 and inhibiting the activation of TGF-β/Smad signaling pathway.

To investigate the effect of alpinetin （ALP） in the letrozole-induced polycystic ovary syndrome （PCOS） model rats， and to elucidate its potential mechanism.

The SD rats were randomly divided into control group， model group， low dose of ALP group and high dose of ALP group.The PCOS rat models were constructed by oral administration of 1 mg·kg^－1·d^－1 letrozole for 21 d， and the rats in low and high doses of ALP groups were orally administrated with 50 and 100 mg·kg^－1 ALP once a day for 14 consecutive days after modeling. The body weights， ovary weight/body weight ratios and oestrous cycle of the rats in various groups were detected. The levels of testosterone and 17β-estradiol in serum of the rats were detected by ELISA. The pathomorphology of ovary tissue of the rats in various groups was observed by HE staining. The expression intensities and distribution of Cleaved cysteine-containing aspartate-specific protease-3 （Cleaved caspase-3） protein in ovary tissue of the rats in various groups were observed by immunohistochemical staining.The expression levels of B-cell lymphoma-2（Bcl-2）， Bcl-2 associated X protein （Bax）， and microtubule associated protein 1 light chain 3 （LC3）-Ⅱ（LC3-Ⅱ）and LC3-Ⅰ proteins in granulosa cells of the rats in various groups were detected by Western blotting method.The Bax/Bcl-2 and LC3-Ⅱ/LC3-Ⅰ ratios were calculated.

Compared with control group， the body weight， the ovary weight/body weight ratio， and the percentage of diestrous phase of the rats in model group were increased（P<0.01），and the levels of testosterone and 17β-estradiol in serum were increased （P<0.01）； compared with model group， the body weights， the ovary weight/body weight ratios， the percentage of diestrous phase and the serum testosterone level in low and high doses of ALP groups were significantly decreased（P<0.05 or P<0.01）， and the serum 17β-estradiol levels in high dose of ALP group were significantly decreased（P<0.05）. The structure of granulosa cell layer in ovary of the rats in control group was normal； the ovary of the rats in model group showed polycystic changes， the fluid-filled large cystic follicles and atretic follicles were found， and the granulosa cell layer was disordered and the number of layers was reduced； the number of cystic follicles and atretic follicles in ovary of the rats in low and high doses of ALP groups was decreased， the structure of granulosa cell layer tended to be normal.The expression of Cleaved caspase-3 in the granulosa cell layer of the rats in control group；the expression of Cleaved caspase-3 in granulose cell layer of the rats in model group was posititive，and the expression intensities of Cleaved caspase-3 in granulosa cell layer in low and high doses of ALP groups were decreased compared with model group.Compared with control group， the ratios of Bax/Bcl-2 and LC3-Ⅱ/LC3-Ⅰ in granulosa cells of the rats in model group were significantly increased （P<0.01）； compared with model group， the ratios of Bax/Bcl-2 and LC3-Ⅱ/LC3-Ⅰ in granulosa cells of the rats in low and high doses of ALP groups were significantly decreased （P<0.05 or P<0.01）.

ALP exerts the therapeutical effects in the letrozole-induced PCOS model rats which might be due to its ability to decrease the levels of testosterone and 17β-oestradiol in the peripheral blood and the levels of autophagic death-related proteins in granulosa cells.

To construct the lentiviral vector of lymphocyte activation gene 3 （LAG3）-mCherry red fluorescent protein （LAG3-mCherry） and establish the cell line stably expressed LAG3-mCherry after transfection of the HEK293T cells， and to investigate the cellular location of LAG3-mCherry fusion protein in the HEK293T cells.

The LAG3 plasmid and lentiviral vector including mCherry gene were digested by endonucleases EcoRⅠ and NotⅠ. The insert and vector fragments were extracted and ligated to construct the pEZ-LAG3-mCherry plasmid. The pEZ-LAG3-mCherry plasmid was sequenced and transfected into the HEK293T cells using Lipofectamine 3000. The expression location of LAG3-mCherry in the HEK293T cells was observed with fluorescence microscope and the expression of LAG3-mCherry fusion proteins in the cells were detected by Western blotting method.

The results of double digestion of recombinant plasmid showed that two DNA bands of about 8 012 and 2 066 bp were found in gel electrophoresis， which were consistent with the size of pEZ-Lv-mCherry vector and LAG3 gene fragment. The DNA sequencing results showed that the LAG-3 gene was successfully inserted into the lentiviral expression vector. The fluorescence microscope results demonstrated that most of LAG3-mCherry proteins were located on the cell membrane of HEK293T cells， while a few fusion proteins were found in the cytoplasm area. The results of Western blotting method showed a specific protein band of LAG3 in the HEK293T cells transfected with pEZ-LAG3-mCherry plasmid.

The pEZ-LAG3-mCherry plasmid including LAG3-mCherry DNA sequence is successfully constructed and the cell line stably expressing LAG3-mCherry is established. The LAG3-mCherry fusion protein is mainly distributed on the cell membrane of HEK293T cells transfected with pEZ-LAG3-mCherry plasmid.

To explore the prognostic value of thioredoxin domain containing 11（TXNDC11） in pan-cancer and its relationship with tumor immune cell infiltration by bioinformatics， and to clarify its evaluation value in tumor prognosis.

The expressions of TXNDC11 mRNA and protein in tumor tissue and corresponding paracancerous tissue were analyzed using ONCOMINE database and TIMER database. The expression difference of TXNDC11 protein between tumor tissue and normal tissue was analyzed by Clinical Proteomic Tumor Analysis Consortium （CPTAC） database. The correlations between the TXNDC11 protein expression and pan-cancer stages were analyzed using Gene Expression Profile Interactive Analysis （GEPIA） database. The relationship between TXNDC11 expression and survival prognosis of patients with pan-cancer was analyzed using online Kaplan Meier plotter. The correlations between the TXNDC11 protein expression and tumor- infiltrating immune cells were studied by TIMER database. The relationship between TXNDC11 protein and immune checkpoint was analyzed using Sangerbox database.

The ONCOMINE database analysis results showed that compared with paracancerous normal tissue， the expression levels of TXNDC11 mRNA in colon adenocarcinoma（COAD），stomach adenocarcinoma（STAD） and sarcoma（SARC） tissues were decreased significantly （P<0.01）， while the expression levels of TXNDC11 mRNA in breast cancer（BRCA），esophageal carcinoma（ESCA）， kidney chromophobe（KICH） and pancreatic adenocarcinoma（PAAD） tissues were increased （P<0.01）. In TIMER database，the expression levels of TXNDC11 protein in BRCA， cholangiocarcinoma（CHOL）， glioblastoma multiforme（GBM）， head and neck squamous cell carcinoma（HNSC） and prostate adenocarcinoma（PRAD） tissues were increased，and the TXNDC11 protein expression levels in COAD，kidney renal papillary cell carcinoma（KIRP），lung adenocarcinoma（LUAD）， lung squamous cell carcinoma（LUSC），pheochromocytoma and paraganglioma（PCPG），rectum adenocarcinoma（READ） and thyroid carcinoma（THCA） tissues were signifcantly decreased.The CPTAC database analysis showed that the protein expression levels of TXNDC11 in kidney renal clear cell carcinoma（KIRC） and uterine corpus endometrial carcinoma（UCEC） were significantly higher than those in normal tissue （P<0.01），but theexpression levels of TXNDC11 protein in BRCA，ovarian cancer（OV） and COAD tissues were lower than those in normal tissue （P<0.05）.The expression level of TXNDC11 protein in GEPIA database was closely related to the clinical characteristics and pathological stages of LUAD，ovarian serous cystadenocarcinoma， THCA and UCEC patients（P<0.05）.In Kaplan Meier plotter database， the high expressions of TXNDC11 in bladder urothelial carcinoma （BLCA） （HR=0.71，P=0.024）， BRCA（HR=0.68， P=0.021）， ESCA （HR=0.39，P=0.005 1），HNSC （HR=0.72，P=0.026）， KIRC（HR=0.41，P=0.002 6），LUAD（HR=0.53，P<0.01），READ（HR=0.34，P=0.014），SARC（HR=0.61，P=0.016），thymoma （THYM）（HR=0.14， P=0.004 2），THCA（HR=0.35，P=0.043） and UCEC（HR = 0.36， P<0.01） tissues were associated with the better prognosis of the patients. The TIMER database analysis results showed that TXNDC11 was significantly correlated with the levels of B cell infiltration in 22 kinds of tumor tissues（P<0.05）；in addition， the expression of TXNDC11 was significantly correlated with the infiltration levels of CD4 + T cells in 16 kinds of tumor tissues，CD8 +^{T cells in 25 kinds of tumor tissues， neutrophils in 22 kinds of tumor tissues，macrophages in 22 kinds of tumor tissues and the dendritic cells in 23 kinds of tumor tissues （P<0.05）；TXNDC11 protein expression was significantly correlated with immune cells in COAD， LUSC， HNSC， SARC and skin melanoma （SKCM）（P<0.01）.The Sangerbox database analysis results showed that TXNDC11 protein was significantly correlated with the expression levels of multiple immune checkpoint genes in a variety of tumor tissues （P<0.05）.}

In pan-cancer analysis， TXNDC11 is generally low expressed in a variety of tumor tissue， and TXNDC11 is significantly correlated with the prognosis and survival of patients with a few of tumors. Therefore， TXNDC11 can be used as an important predictor of tumor prognosis and regulate the various tumor-related immune responses in a certain degree.

To construct the potential miRNA-mRNA regulatory network of fusion genes in the pathogenesis of rhabdomyosarcoma （RMS）with bioinformatics analysis， and to provide a new direction for the study of RMS.

GEO2R analysis tool was used to screen the differentially expressed genes （DEGs） and differentially expressed miRNA in fusion-gene-positive and -negative RMS tissues； the target genes of differentially expressed miRNA were predicted； and the target genes on the basis of overlapping DEGs and target genes were screened out. DAVID database was implemented for the GO and KEGG enrichment analysis of target genes. STRING and Cytoscape software were utilized to construct the protein protein interation （PPI） network， the Top10 hub genes were screened out and the molecular regulation networks of hub genes and miRNA were constructed， and the Kaplan-Meier survival curves of the sarcoma patients were drawn.

A total of 891 DEGs （P<0.01，|logFC|≥1） and 14 differentially expressed miRNAs （P<0.05， |logFC|≥3） were screened out. Moreover， 1 654 target genes differentially expressing miRNAs were predicted， and 115 target genes were identified by overlapping the target genes and DEGs. The GO enrichment analysis revealed that the target genes were mainly enriched in the positive regulation of cell proliferation， cell surface， protein binding， and other biological processes. The KEGG signaling pathway analysis revealed that the target genes were mainly enriched in the extracellular matrix-receptor interaction， proteoglycan in cancer， and other pathways. The Top10 hub genes screened out from the PPI network were ERBB4， PTPRD， IRS1， ADAM10， YAP1， TFAP2A， CADM1， ELAVL2， SNAI1， and ERRFI1. The survival analysis showed that the increased expression levels of PTPRD （HR=1.79， P=0.005），ADAM10 （HR =1.94，P=0.010）， ELAVL2 （HR = 1.56，P=0.031），and ERRFI1 （HR=2.05，P=0.005） were related to the bad prognosis of the sarcoma patients.

The miRNA-mRNA network is constructed by selecting the hub genes and corresponding miRNA involed in the pathogenesis of RMS， and the study provides a theoretical basis for further study of the relationship between fusion genes and RMS.

To analyze the expressions of matrix metalloproteinase-9 （MMP-9） and tissue inhibitor of metalloproteinase-1 （TIMP-1） in cancer tissue of the gastric cancer patients， and to investigate their values in prognostic evaluation of the gastric cancer patients underwent radical gastrectomy.

A total of 270 gastric cancer patients who underwent radical gastrectomy were enrolled in this study， and the positive expression rates of MMP-9 and TIMP-1 in cancer tissue of the patients were detected by immunohistochemistry method. Pearson χ² or Fisher’s exact test was used to assess the distribution of MMP-9 and TIMP-1 positive expressions in the gastric cancer patients with different clinicopathological parameters. Survival curves were plotted using Kaplan-Meier method and the differences were assessed by Log-rank test. Multivariate Cox regression model was conducted to analyze the relationships between the expressions of MMP-9 and TIMP-1 and the prognosis of the gastric cancer patients underwent radical gastrectomy.

Among the 270 gastric cancer patients， MMP-9 and TIMP-1 were expressed in cytoplasm or cell membrane of tumor cells， and the positive expression rates of MMP-9 and TIMP-1 were 30.4% and 26.3%， respectively. No correlations between the co-expression of MMP-9， TIMP-1 and the age，gender，tumor size，tumor type，histological type，chemotherapy after operation，and TNM stage of the patients were identified. The results of Cox regression model showed that co-expression of MMP-9 and TIMP-1， postoperative chemotherapy and TNM stage were the significant independent factors for the prognosis of the gastric cancer patients. The gastric cancer patients with MMP-9 positive expression showed the better survival， and among the patients withMMP-9 negative expression， the risk of death in the patients with TIMP-1 positive expression was increased， which indicated the worse prognosis（MMP-9⁺/TIMP-1^+/－vs MMP-9^－/TIMP-1⁺：HR=0.537， 95% CI： 0.338－0.851，P=0.008）. Furthermore， compared with the patients with postoperative chemotherapy and TNM stage Ⅰ/Ⅱ， the patients with no-postoperative chemotherapy and TNM stage Ⅲ had higher risk of death and worse prognosis（HR=1.873，95% CI： 1.334－2.631，P<0.001； HR=2.301， 95% CI： 1.656－3.197， P<0.001）.In the subgroup analysis， among the gastric cancer patients with TNM stage Ⅰ/Ⅱ， postoperative chemotherapy or no signet-ring cell cancers， the risk of death in MMP-9⁺ group was decreased compared with MMP-9^－/TIMP-1⁺ subgroup （P<0.05），which indicated the better prognosis.

Co-expression of MMP-9 and TIMP-1 could serve as an independent prognostic predictor in the gastric cancer patients underwent radical gastrectomy； No postoperative chemotherapy and TNM stage Ⅲ are the risk factors for the prognosis of gastric cancer patients underwent radical gastrectomy.

To explore the expression of suppressor of cytokine signaling 3 （SOCS3） in the peripheral blood mononuclear cells of the patients with diffuse large B-cell lymphoma （DLBCL） and its effect on the autophagy and apoptosis of OCI-LY1-cells， and to clarify its molecular mechanism.

The peripheral blood mononuclear cells of healthy volunteers （control group， 50 cases） and DLBCL patients （DLBCL group， 100 cases） were collected and extracted，and the human B lymphocytes and OCI-LY7 cells were cultured at the same time. The expression levels of SOCS3 mRNA in peripheral blood mononuclear cells， B lymphocytes，and OCI-LY7 cells of the subjects in two groups were detected by real-time fluorescence quantitative PCR （RT-qPCR） method. The OCI-LY7 cells were divided into pcDNA3.1-NC group and pcDNA3.1-SOCS3 group， and the cells were transfected with pcDNA3.1-NC and PCDNA3.1-SOCS3， respectively.The expression levels of SOCS3 mRNA in the cells in two groups were detected by RT-qPCR method；the EDU positive cell rates of the cells in two groups were detected by EDU experiment， and the microtubule-associated protein 1 light chain 3（LC3） positive cell rates in the cells in two groups were detected by cell immunofluorescence staining； flow cytometry was used to detect the apoptotic rates and the percentages of cells in different cell cycles in two groups，and Western blotting method was used to detect the expression levels of SOCS3， phosphorylated Janus kinase 2（p-JAK2） and phosphorylated signal transducer and activator of transcription 3 （p-STAT3） proteins in the cells in two groups.The ratios of LC3Ⅱ/LC3Ⅰ was calculated.

Compared with control group， the expression level of SOCS3 mRNA in peripheral blood mononuclear cells of the patients in DLBCL group was decreased （P<0.01）； compared with B lymphocyte group， the expression level of SOCS3 mRNA in the OCI-LY7 cells was decreased （P<0.01）. Compared with pcDNA3.1-NC group，the expression levels of SOCS3 mRNA and protein in the cells in pcDNA3.1-SOCS3 group were significantly increased（P<0.01）， the EDU positive cell rate of OCI-LY7 cells was significantly decreased （P<0.01）， the ratio of LC3 Ⅱ/LC3Ⅰ and the number of LC3 positive cells were decreased significantly（P<0.01），the apoptotic rate of OCI-LY7 cells was significantly increased （P<0.01）， the percentage of cells in G₀/G₁ phase was significantly increased （P<0.01）， the percentage of cells in S phase was significantly reduced （P<0.01），and the expression levels of p-JAK2 and p-STAT3 proteins were significantly reduced （P<0.01）.

SOCS3 is lowly expressed in the peripheral blood mononuclear cells and OCI-LY7 cells of the patients with DLBCL. Overexpression of SOCS3 can inhibit the proliferation and autophagy of OCI-LY7 cells， and enhance the apoptotic rate， and its mechanism may be related to inhibiting the JAK2/STAT3 signaling pathway.

To examine the dynamic changes in BCR/ABL fusion gene expression in peripheral blood in the patients with Ph⁺ acute lymphoblastic leukemia （Ph⁺ALL） by real-time fluorescence quantitative PCR（RT-qPCR），and to formulate the different treatment measures for different patients and improve the overall survival（OS） rates of patients.

The clinical materials of twenty patients with newly treated and refractory recurrent Ph⁺ALL were analyzed retrospectively. After induction treatment with tyrosine kinase inhibitor+vincristine+prednisone（TKI+VP） or tyrosine kinase inhibitor+vincristine+daunorubicin+prednisone（TKI+VDP），17 patients achieved hematological complete remission（HCR） and 3 patients did no remission （NR）. Among the HCR patients， 8 patients for minimal residual disease （MRD） were negative， 3 patients chose Auto-HSCT， and all received TKI maintenance treatment till now after hematopoietic function reconstruction； Allogeneic hematopoietic stem cell transplantation （Allo-HSCT） was performed in other 5 MRD negative patients， 9 MRD positive patients and 3 NR patients.The BCR/ABL fusion genes in the peripheral blood of the patients were detected before transplantation，1 month and 2，3，6，9，and 12 months after stem cell transplantation，and the MRD of the patients was evaluated.After hematopoietic reconstruction in Allo-HSCT group， TKI oral intervention was given regardless of the results of BCR/ABL gene quantitative detection. TKI oral intervention was discontinued after one year for those who continued to test negative. If positive MRD conversion was monitored during the period， effective TKI drugs or immunosuppressant reduction， and other therapeutic measures were given according to the time of transplantation.

All patients achieved hematopoietic reconstruction after transplantation， the median survival time of granulocyte transplantation was 14 （11－24） d， and the median survival time of platelet transplantation was 17 （13－52） d. Follow-up was conducted until December 2020， with a median follow-up time of 52 （3－111） months.The 3-year overall survival （OS） rates of all the patient was （61.1±11.7）%. The 3-year OS rates in Allo-HSCT group and Auto-HSCT group were （52.8±13.4）% and 100.0% （P=0.178），respectively， and the difference was not statistically significant（P=0.178）.The 3-year OS rates of the patients in MRD negative group before treatment and MRD positive group with HCR before treatment were （87.5±11.7）% and （42.8± 15.6）% （P=0.065）， respectively，and the difference was not statistically significant（P=0.065）.All patients with NR before transplantation died.Five cases in whole group died of disease recurrence，1 case of lung infection and 1 case of grade Ⅳ acute graft-versus-host disease.

In the patients with Ph⁺ALL， choosing autologous or allogeneic hematopoietic stem cell transplantation and postoperative stratified intervention according to the dynamic changes of BCR/ABL fusion gene results can improve the OS rate of patients.

To screen the differentially expressed genes （DEGs） of endometriosis （EMS） by bioinformatics tools，and to provide theoretical basis for clarifyinng the pathogenesis of EMS， and to find a new research direction for the diagnosis and treatment of EMS.

The datasets （GSE25628， GSE23339 and GSE7305） including EMS tissue and normal endometrial tisue were downloaded from the Gene Expression Omnibus（GEO） database by searching for endometriosis. The GE02R online analvsis tools were used to screen for the DEGs in the EMS tissue and the normal endometrial tissue. Gene Ontology （GO） enrichment analysis and Kyoto Encyclopedia of Genes and Genomes （KEGG） were used to analyze the function and pathway enrichment of DEGs， the STRING database was used to establish the protein-protein interaction （PPI） network. Finally，the PPI network was analyzed and visualized by Cytoscape software， and the top ten core genes were screened.

After the DEGs analysis， a total of 27 up-regulated DEGs and 43 down-regulated DEGs were identified. The GO enrichment analysis results showed that DEGs were mainly enriched in the extracellular matrix， extracellular space， extracellular region and so on. The results of KEGG pathway analysis demonstrated that DEGs were mainly enriched in phosphate inositol metabolic signal pathway.Through STRING and Cytoscape softwares， the top 10 core genes with the highest connectivities in the PPI network were screened out， including DCN， EPCAM， BGN， FABP4， NTRK2， GPC-3， ERBB3， CD24， NID2， and THBS2. Among them， DCN， BGN， NID2， and THBS2 were associated with the biological function of extracellular matrix.

The disorder of extracellular matrix-related genes may play an important role in the occurrence and development of EMS， and the extracellular matrix-related genes may become the biomarkers and for early diagnosis and therapeutic targets of EMS.

To investigate the effect of combined detection of the biomarkers human mammaglobin （hMAM）， small breast epithelial mucin （SBEM） and carcino-embryonic antigen related adhesion molecule 19 （CEACAM19） mRNA in the diagnosis of breast cancer， and to clarify its clinical significance.

The peripheral blood was collected from 104 breast cancer patients（breast cancer group）， 52 benign breast disease patients（benign breast disease group） and 50 healthy individuals（healthy control group）， and the expression levels of hMAM， SBEM and CEACAM19 mRAN in peripheral blood nucleated cells were detected by real-time fluorescence quantitative PCR（RT-qPCR） method； the positive expression rates were calculated and their relationships with patients’ clinicopathologic features were analyzed.

In breast cancer group， the positive expression rates of SBEM and CEACAM19 mRNA in peripheral blood were associated with lymph node metastasis （P<0.05）， the positive expression rate of CEACAM19 mRNA was associated with estrogen receptor（ER） expression （P<0.05）， and the positive expression rate of hMAM mRNA was associated with human epidermal growth factor receptor-2（HER-2） expression（P<0.05）. The positive expression rates of hMAM mRNA in peripheral blood in breast cancer group were significantly increased compared with healthy control group and benign breast disease group（P<0.05）， both alone and in combined with SBEM and CEACAM19. The positive expression rate of hMAM mRNA was increased from 39.4% to 66.3% by combined detection of the three indicators. The positive expression rates of hMAM mRNA in peripheral blood of the breast cancer patients at TNM stage Ⅲ+Ⅳ were higher than those of patients at TNM stage Ⅰ+Ⅱ（P<0.01）， both alone and in combined with SBEM and CEACAM19. By combined detection of hMAM，SBEM， andCEACAM19 mRNA， the positive expression rate of hMAM mRNA in peripheral blood of the breast cancer patients at TNM stage Ⅰ+Ⅱwas increased from 20.40% to 48.98% and at TNM stage Ⅲ+Ⅳ from 56.40% to 81.82%.

The positive expression rate of hMAM mRNA in peripheral blood of the breast cancer patients is not high. Combined detection of SBEM and CEACAM19 mRNA can increase the positive expression rate， which has certain clinical significance in the diagnosis of breast cancer.

To detect the coagulation indexes， platelet count and neutrophil/lymphocyte ratio （NLR） in the peripheral blood of patients with malignant tumors before clinical treatment， and to explore their evaluation values in the patient’s coagulation state.

A total of 354 patients with malignant tumors（74 cases of cervical cancer， 65 cases of lung cancer， 55 cases of ovarian cancer， 50 cases of colorectal cancer， 43 cases of gastric cancer， 35 cases of esophageal cancer and 32 cases of pancreatic cancer） were selected as tumor group and divided into early stage （Ⅰ-Ⅱ） and late stage （Ⅲ-Ⅳ） tumor groups according to TNM stage. At the same time， 100 healthy patients were selected as control group. The prothrombin time （PT）， activated partial thromboplastin time （APTT）， thrombin time （TT），fibrinogen （FIB）， D-dimer （D-Dimer）， fibrin or fibrinogen degradation products （FDP）， antithrombin （AT）， platelet count and NLR of the subjects in various groups were compared and analyzed.

Compared with control group， PT in early and late stage tumor groups were prolonged （P<0.01）， APTT were shortened （P<0.05 or P<0.01）， and FIB， D-Dimer， FDP levels and NLR were all increased （P<0.01）；compared with control group， TT of the patients in late tumor group was shortened （P<0.05）， AT was decreased （P<0.05）， and platelet count was increased （P<0.05）；compared with early stage tumor group， PT of the patients in late tumor group was prolonged （P<0.01）， TT was shortened （P<0.01）， and FIB， D-Dimer， FDP levels and NLR were increased （P<0.01）.Compared with control group， PT of the patients in all tumor groups were prolonged （P<0.05 or P<0.01）， APTT were shortened in different types of tumor groups except esophageal cancer （P<0.05 or P<0.01）， TT of the patients with lung cancer， ovarian cancer， and esophageal cancer were shortened（P<0.05 or P<0.01）， FIB， D-Dimer and FDP in all tumor groups were increased（P<0.05 or P<0.01）， AT of the patients with colorectal cancer， esophageal cancer， and pancreatic cancer were decreased （P<0.05）， the platelet count of the patients with advanced ovarian cancer was increased （P<0.01）， and NLR in all tumor groups were increased（P<0.05 or P<0.01）.

In the patients with malignant tumors before treatment， the main indicators of peripheral blood are shortened APTT， increased FIB and D-Dimer levels and NLR， and the patients with advanced tumors also show increased platelet count and decreased AT， indicating that tumor patients have disorder of hemostasis and coagulation function and are prone to thrombotic events.

To analyze the clinical manifestations of secondary glaucoma caused by varicella-zoster virus（VZV） keratouveitis， and to explore its treatment plan.

The clinical information of a patient with secondary glaucoma caused by VZV keratouveitis was collected，and the relevant literatures were reviewed. The clinical symptoms， diagnosis， and treatment of secondary glaucoma caused by VZV keratouveitis were analyzed.

The female patient aged 55 years old was admitted to hospital because of “visual acuity of the left eye decreased for more than 40 d”. Physical examination showed conjunctival congestion in the left eye， diffuse corneal edema， endothelial pigment keratic precipitates（KP）， aqueous humor turbidity and glow， iris depigmentation， part of iris posterior synechia， and fundus blurred. The gonioscope results showed a large amount of pigmentation in the chamber angle， and aqueous humor virus detection showed that VZV was strongly positive， which was diagnosed as secondary glaucoma caused by VZV keratouveitis. The patient was locally treated with systemic anti-inflammatory， antiviral and intraocular pressure-lowering drugs， and the intraocular pressure was decreased insignificantly.After anterior chamber irrigation， the intraocular pressure was decreased， and the condition was improved and the patient was discharged from the hospital.

Aqueous virus testing is an effective method for diagnosing VZV keratouveitis， anterior chamber washout is a successful treatment for glaucoma caused by VZV keratouveitis with a lot of pigmentation in the corners.

To analyze the clinical symptoms， laboratory examination and ¹⁸F-FDG PET/CT imaging of a patient with primary small cell osteosarcoma of coccyx with concurrent metastases， and to improve the physician’s diagnosis of rare manifestation for this common disease.

The patient’s clinical data of the patient with primary small cell osteosarcoma of coccyx with concurrent metastases were collected， and its clinical characteristics， imaging performance， diagnosis and treatment methods were analyzed through the review of relevant literatures.

A 10-year old girl admitted to hospital due to neck pain for two weeks and fever for one week， and a mass in lower right lung lobe found in lung CT examination. Body examination showed body temperature up to 37.5 ℃， no other discomfort. The laboratory results indicated alkaline phosphatase 221.3 U·L^－1， creatinine 38.1 μmol·L^－1.The ¹⁸F-FDG PET/CT results showed high metabolic mass in the lower lobe of the right lung， considering malignancy； the metabolism of the left side of the cervical 2 vertebrae was increased， maybe metastatic tumors； the tail bone density was not equal， a high metabolism soft tissue nodule protruding into the pelvis， considering malignant neoplasm.The MRI results showed that the axoid morphology was irregular， with long T1 and T2 signals， and the lesion was markedly enhanced； slightly longer T1 and slightly longer T2 signals lesions were found in sacral 5 level spinal canal， sacral 5 vertebral body and caudal vertebrae and around the capsule， and no obvious enhancement； anterior soft tissue around the sacral vertebrae anterior soft tissue was enhanced， space occupying lesion was considered. The postoperative pathology of the lower lobe of the right lung was considered metastatic small cell-type osteosarcoma for the lungs.Based on the above manifestations and pathological analysis， puncture biopsy of soft tissue lesions of the coccyx was performed， and the pathological result indicated small cell osteosarcoma. Finally the patient was diagnosed as primary small cell osteosarcoma of coccyx combined with lung and bone metastasis.

Primary tail bone small cell osteosarcoma is difficult to be diagnosed， but ¹⁸F-FDG PET/CT molecular imaging plays a key role in clarifying the lesion scope，tumor metabolic degree， and guiding the biopsy site.

To analyze the clinical manifestations， diagnostic methods and treatment process of the patients with Chlamydia psittaci pneumonia and improve the understanding of clinicians， and improve the efficiency of clinical diagnosis and treatment and the prognosis of the disease.

The clinical data of 2 patients with Chlamydia psitsiti pneumonia were collected， and the diagnosis and treatment process was summarized；the infection indicators， oxygenation function， dynamic changes of pulmonary imaging were analyzed；culture method，serological method， metagenomic next-generation sequencing technology （mNGS） and other etiological diagnostic methods were compared，and the effects of quinolones and tetracycline in the treatment of Chlamydia psittaci pneumonia were analyzed.

The first symptoms of two patients were fever， cough and expectum， and the disease progressed rapidly. Dyspnea developed on the 8th day and the 3rd day of onset， respectively. One patient developed acute respiratory distress syndrome（ARDS） on the 10th day of onset， and mechanical ventilation assisted treatment was used. The counts of white blood cells in blood routine examination of two patients were 12.69×10⁹·L^－1 and 4.4 ×10⁹·L^－1， and procalcitonin were 0.27 and 0.06 μg·L^－1， respectively. Routine culture of airway secretes and serological examination failed to identify the pathogen. The initial diagnosis of two patients was community acquired pneumonia， and the effect of empirical treatment with quinolones was not good. The mNGS method combined with bird contact history was used to finally confirm the diagnosis of Chlamydia psittaci pneumonia.The symptoms， signs and imaging abnormalities were significantly improved after treatment with tegecycline， a tetracycline antibiotic. The results of lung CT reexamination of 2 cases after 1 month follow-up showed that the lesions were basically absorbed.

The occurrence of Chlamydia psittaci pneumonia is related to the contact history of birds. The progress of the disease is often rapid. The mNGS technology can be used as a rapid etiological diagnosis method， which is conducive to the early diagnosis of Chlamydia psittaci pneumonia， the timely selection of sensitive antibiotics and the improvement of disease prognosis. A new generation of tetracyclines represented by doxycycline and tigecyclin has a positive effect on Chlamydia psittaci pneumonia.

To explore the clinical characteristics， diagnosis， treatment and prognosis of the children with Kawasaki disease complicated with inflammatory changes in parapharyngeal space and retropharyngeal space， and to improve the clinicians’ understanding of the disease.

The clinical data of one case of Kawasaki disease complicated with parapharyngeal space involvement as the main manifestation hospitalized in our department in November 2019 were retrospectively analyzed，and the clinical manifestations and imaging characteristics of Kawasaki disease complicated with head and neck symptoms were analyzed by reviewing and summarizing the previous literatures.

The male 3-year old patient had fever for 8 d，and was hospitalized due to neck movement limitation and torticula for 2 d. The physical examination results showed that there were red macular papules on the trunk， swollen lymph nodes on the neck， conjunctival hyperemia， red lips， waxberry tongue， and hard swelling of hands and feet. The total number of white blood cells and C-reactive protein （CRP） in blood routine examination were significantly increased. On the 20th day of the course of disease， typical membranous desquamation appeared in the extremities of the patient， which was clearly diagnosed as Kawasaki disease.The children had neck movement limitation and torticollis on admission，and the imaging results suggested a mass in the right parapharyngeal space of nasopharynx and oropharynx. The symptomatic treatment of the patient was ineffective after given antibiotics and antipyretic. After treatment with gamma globulin and aspirin， the neck movement limitation and torticula of the patient disappeared， the symptoms and signs of Kawasaki disease were improved， and the inflammatory indicators of laboratory examination were gradually returned to normal， and the neck MRI results showed that the inflammatory range of parapharyngeal space was significantly smaller than before.

The abnormal manifestations of head and neck can be presented as early manifestations of Kawasaki disease， which is easily misdiagnosed as suppurative lymphadenitis or parapharyngeal and retropharyngeal suppurative infection. Kawasaki disease complicated with parapharyngeal space or retropharyngeal space inflammation should be correctly diagnosed and treated as soon as possible to avoid the cardiovascular system damage and unnecessary surgical treatment.

Using data from China disease Control and Prevention Information System，the epidemiological data of the hand-foot-mouth disease were analyzed with descriptive epidemiological method.The throat swab speimens of the patients with hand-foot-mouth disease were collected from Changchun medical institutions from 2014 to 2018.Real-time fluorescence quantitative PCR （RT-PCR） was used to detect the total enterovirus， enterovirus 71（EV-A71） and Coxsackieevirus 16（CV-A16） in the speimens to confirm the types.The VP1 regions of EV-A71 were amplificated and sequenced and the phylogenetic tree was constructed.The homology sites and amino acid variation sites were analyzed.

From 2014 to 2018， a total of 12 685 cases were reported in Changchun City of Jilin Province with the incidence decreasing first and then increasing， reaching a peak in 2016 and decreasing year by year thereafter. The epidemic trend was basically the same， showing a single peak distribution， with obvious seasonal. The cases were reported in 14 counties and districts of Changchun City， and the top three areas with annual incidence were high-tech zone （129.14/100 000）， broad urban area （102.17/100 000） and economic development zone （100.98/100 000）.The proportion of male cases was significantly higher than that of female cases， and the majority of cases were children under 5 years old.The EV-A71 positive samples were detected every month in Changchun City.During the study period from March to September 2014 and from May 2017 to May 2018， EV-A71 was the main causative agent of hand-foot-mouth disease， showing a gradually upward trend.A total of 146 EV-A71 sequences were obtained， 136 of the EV-A71 strains samples belonged to C4a subtypes of subgenogroup C4， whereas 10 of the EV-A71 strains samples belonged to subgenogroup B5 of EV-A71.The position 145 amino acids of VP1 region was E；VP1^104D was found in two strains，and VP1^218R was found in the other three strains in 2018；the other toxicological sites and the main antigenicity sites did not change.Compared with the C4 subtype，B5 subtype had three amino acid differences（VP1^43E，VP1^58Tand VP1^184T）.

Hand-foot-mouth disease is a common infectious disease， which in Changchun City of Jilin Province mainly occurs among the children younger than 5 years old. It is mainly caused by C4a subtype of EV-A71， and a small part of B5 subtype is prevalent， some amino acid variant sites need to be continuously monitored to avoid the pandemics.