Objective:To investigate the expression of SAPCD2 in esophageal squamous cell carcinoma (ESCC) tissues and its effects on the biological function of ESCC cells in vitro, as well as the possible molecular mechanisms.Methods:By using the Gene Expression Profile Interaction Analysis (GEPIA) platform, the transcriptional level expressions of SAPCD2 in 182 ESCC samples and 286 normal esophageal tissue samples from the Cancer Genome Atlas (TCGA) database and Genotype-Tissue Expression (GTEx) database were analyzed. Immunohistochemical (IHC) staining was performed on clinical tissue chips of ESCC patients, and staining scores were evaluated. The expression differences of SAPCD2 protein between 61 cancer tissues and the paired adjacent tissues with the complete clinical data, as well as the distribution of patients with SAPCD2 high expression among patients stratifies by different clinicopathological features were compared. ESCC cell line KYSE-150 was transfected with plasmids carrying SAPCD2 sequence and short hairpin RNA sequence targeting SAPCD2, respectively, which was treated as the SAPCD2 overexpression group and SAPCD2 knockdown group; and the cells transfected with empty plasmids and plasmids carrying negative RNA sequence were treated as the overexpression control group and the knockdown control group. CCK-8 method (expressed with the absorbance value) and plate clone formation assay were used to detect the cell proliferation ability. Cell migration was detected by using cell scratch assay and Transwell cell migration assay were used to detect the cell migration ability. The reverse-real time fluorescence quantitative polymerase chain reaction (qPCR) was used to detect the mRNA levels of SAPCD2, matrix metalloproteinase-9 (MMP9), proliferating cell nuclear antigen (PCNA), and Vimentin in cells of all groups. Western blotting was used to detect the expression levels of SAPCD2 protein and proteins related to cell proliferation, invasion, metastasis, and AKT signaling pathway.Results:GEPIA platform analysis showed that the transcriptional expression level of SAPCD2 in ESCC tissues was higher than that in adjacent tissues, and the difference was statistically significant ( P < 0.01). IHC staining showed that the staining score of SAPCD2 protein in cancer tissues was higher than that in adjacent tissues [(8.2±2.8) points vs. (2.2±1.7) points], and the proportion of patients with positive SAPCD2 protein (staining score > 0 point) in cancer tissues was higher than that in adjacent tissues [95.1% (58/61) vs. 57.4% (35/61)], and the differences were statistically significant (all P < 0.001), while there were no statistically significant differences in the distribution of the high expression of SAPCD2 protein (staining score > 3 points) patients stratified by gender, age, tumor size, pathological grade, and T stage, N stage and M stage (all P > 0.05). CCK-8 assay showed that the absorbance value of KYSE-150 cells in the SAPCD2 overexpression group after 96 h of culture was higher than that in the overexpression control group, while the absorbance values of the SAPCD2 knockdown group after 72 h and 96 h of culture were lower than those in the knockdown control group, and the differences were statistically significant (all P < 0.05). The plate clone formation assay showed that the number of colonies of KYSE-150 cells cultured for 14 d in the SAPCD2 overexpression group was more than that in the overexpression control group [(800±30) vs. (458±47)], and that in the SAPCD2 knockdown group was less than that in the knockdown control group [(52±7) vs. (81±2)], and the differences were statistically significant (all P < 0.05). The cell scratch assay showed that after 24 h of culture, the scratch width of KYSE-150 cells in the SAPCD2 overexpression group was narrower than that in the overexpression control group [(51±9) μm vs. (89±7) μm], while that in the SAPCD2 knockdown group was wider than that in the knockdown control group [(120±22) μm vs. (37±10) μm], and the differences were statistically significant (all P < 0.01). Transwell cell migration assay showed that the migration number of KYSE-150 cells in the SAPCD2 overexpression group was more than that in the overexpression control group [(202±18) vs. (50±14)], and that in the SAPCD2 knockdown group was less than that in the knockdown control group [(227±27) vs. (483±16)], and the differences were all statistically significant (all P < 0.01). qPCR assay showed that the mRNA relative expression levels of MMP9, PCNA and Vimentin in KYSE-150 cells in the SAPCD2 overexpression group were all higher than those in the overexpression control group, while those in the SAPCD2 knockdown group were all lower than those in the knockdown control group, and the differences were statistically significant (all P < 0.05). Western blotting showed that the relative expression levels of PCNA and Vimentin proteins in KYSE-150 cells of the SAPCD2 overexpression group were higher than those of the overexpression control group, while the relative expression levels of epithelial cadherin (E-cad) and cleaved cysteine aspartate protease 3 (CASP3) proteins were lower than those of the overexpression control group; however, the expression levels in SAPCD2 knockdown group showed the opposite results, and the differences were statistically significant (all P < 0.05); the relative expression level of phosphorylated AKT (p-AKT) protein in the SAPCD2 overexpression group was higher than that in the overexpression control group, while that in the SAPCD2 knockdown group was lower than that in the knockdown control group, and the differences were statistically significant (all P < 0.01). Conclusions:SAPCD2 is highly expressed at both the transcriptional level and the protein level in ESCC tissues. SAPCD2 promotes the proliferation and migration of ESCC cells in vitro, which may be related to the AKT signaling pathway.

Objective To investigate the impact of intergenerational caregiving on the brain structure and function of grandparents,and to analyze its correlation with caregiving factors.Methods Healthy adults(66 with grandchildren,24 without grandchildren)were recruited as study subjects,and clinical and MRI data were collected.Resting-state brain functional degree centrality(DC)and surface-based morphometry(SBM)methods were used to compare the differences in brain structure and function between the groups with and without grandchildren.The correlation between the differences in brain regions and △ values with grandchild's age,number,and time spent in childcare were assessed,respectively.Results Compared to the group without grandchildren,the group with grandchildren showed reduced surface area and cortical volume in the left middle temporal gyrus,as well as decreased DC values in the left medial superior frontal gyrus,bilateral orbital superior frontal gyrus,and left anterior cingulate and paracingulate gyrus(P＜0.05),respectively.In the grandchildren group,DC values and △ values in the left orbital superior frontal gyrus,left anterior cingulate and paracingulate gyrus were significantly positively correlated with time spent in childcare.Conclusion The brain structures and functions of grandparents related to empathy and motivation are changed in intergenerational caregiving,which may reveal the neuroplasticity after caring for their grandchildren.

Objective To investigate the impact of intergenerational caregiving on the brain structure and function of grandparents,and to analyze its correlation with caregiving factors.Methods Healthy adults(66 with grandchildren,24 without grandchildren)were recruited as study subjects,and clinical and MRI data were collected.Resting-state brain functional degree centrality(DC)and surface-based morphometry(SBM)methods were used to compare the differences in brain structure and function between the groups with and without grandchildren.The correlation between the differences in brain regions and △ values with grandchild's age,number,and time spent in childcare were assessed,respectively.Results Compared to the group without grandchildren,the group with grandchildren showed reduced surface area and cortical volume in the left middle temporal gyrus,as well as decreased DC values in the left medial superior frontal gyrus,bilateral orbital superior frontal gyrus,and left anterior cingulate and paracingulate gyrus(P＜0.05),respectively.In the grandchildren group,DC values and △ values in the left orbital superior frontal gyrus,left anterior cingulate and paracingulate gyrus were significantly positively correlated with time spent in childcare.Conclusion The brain structures and functions of grandparents related to empathy and motivation are changed in intergenerational caregiving,which may reveal the neuroplasticity after caring for their grandchildren.

Objective:To investigate the expression of SAPCD2 in esophageal squamous cell carcinoma (ESCC) tissues and its effects on the biological function of ESCC cells in vitro, as well as the possible molecular mechanisms.Methods:By using the Gene Expression Profile Interaction Analysis (GEPIA) platform, the transcriptional level expressions of SAPCD2 in 182 ESCC samples and 286 normal esophageal tissue samples from the Cancer Genome Atlas (TCGA) database and Genotype-Tissue Expression (GTEx) database were analyzed. Immunohistochemical (IHC) staining was performed on clinical tissue chips of ESCC patients, and staining scores were evaluated. The expression differences of SAPCD2 protein between 61 cancer tissues and the paired adjacent tissues with the complete clinical data, as well as the distribution of patients with SAPCD2 high expression among patients stratifies by different clinicopathological features were compared. ESCC cell line KYSE-150 was transfected with plasmids carrying SAPCD2 sequence and short hairpin RNA sequence targeting SAPCD2, respectively, which was treated as the SAPCD2 overexpression group and SAPCD2 knockdown group; and the cells transfected with empty plasmids and plasmids carrying negative RNA sequence were treated as the overexpression control group and the knockdown control group. CCK-8 method (expressed with the absorbance value) and plate clone formation assay were used to detect the cell proliferation ability. Cell migration was detected by using cell scratch assay and Transwell cell migration assay were used to detect the cell migration ability. The reverse-real time fluorescence quantitative polymerase chain reaction (qPCR) was used to detect the mRNA levels of SAPCD2, matrix metalloproteinase-9 (MMP9), proliferating cell nuclear antigen (PCNA), and Vimentin in cells of all groups. Western blotting was used to detect the expression levels of SAPCD2 protein and proteins related to cell proliferation, invasion, metastasis, and AKT signaling pathway.Results:GEPIA platform analysis showed that the transcriptional expression level of SAPCD2 in ESCC tissues was higher than that in adjacent tissues, and the difference was statistically significant ( P < 0.01). IHC staining showed that the staining score of SAPCD2 protein in cancer tissues was higher than that in adjacent tissues [(8.2±2.8) points vs. (2.2±1.7) points], and the proportion of patients with positive SAPCD2 protein (staining score > 0 point) in cancer tissues was higher than that in adjacent tissues [95.1% (58/61) vs. 57.4% (35/61)], and the differences were statistically significant (all P < 0.001), while there were no statistically significant differences in the distribution of the high expression of SAPCD2 protein (staining score > 3 points) patients stratified by gender, age, tumor size, pathological grade, and T stage, N stage and M stage (all P > 0.05). CCK-8 assay showed that the absorbance value of KYSE-150 cells in the SAPCD2 overexpression group after 96 h of culture was higher than that in the overexpression control group, while the absorbance values of the SAPCD2 knockdown group after 72 h and 96 h of culture were lower than those in the knockdown control group, and the differences were statistically significant (all P < 0.05). The plate clone formation assay showed that the number of colonies of KYSE-150 cells cultured for 14 d in the SAPCD2 overexpression group was more than that in the overexpression control group [(800±30) vs. (458±47)], and that in the SAPCD2 knockdown group was less than that in the knockdown control group [(52±7) vs. (81±2)], and the differences were statistically significant (all P < 0.05). The cell scratch assay showed that after 24 h of culture, the scratch width of KYSE-150 cells in the SAPCD2 overexpression group was narrower than that in the overexpression control group [(51±9) μm vs. (89±7) μm], while that in the SAPCD2 knockdown group was wider than that in the knockdown control group [(120±22) μm vs. (37±10) μm], and the differences were statistically significant (all P < 0.01). Transwell cell migration assay showed that the migration number of KYSE-150 cells in the SAPCD2 overexpression group was more than that in the overexpression control group [(202±18) vs. (50±14)], and that in the SAPCD2 knockdown group was less than that in the knockdown control group [(227±27) vs. (483±16)], and the differences were all statistically significant (all P < 0.01). qPCR assay showed that the mRNA relative expression levels of MMP9, PCNA and Vimentin in KYSE-150 cells in the SAPCD2 overexpression group were all higher than those in the overexpression control group, while those in the SAPCD2 knockdown group were all lower than those in the knockdown control group, and the differences were statistically significant (all P < 0.05). Western blotting showed that the relative expression levels of PCNA and Vimentin proteins in KYSE-150 cells of the SAPCD2 overexpression group were higher than those of the overexpression control group, while the relative expression levels of epithelial cadherin (E-cad) and cleaved cysteine aspartate protease 3 (CASP3) proteins were lower than those of the overexpression control group; however, the expression levels in SAPCD2 knockdown group showed the opposite results, and the differences were statistically significant (all P < 0.05); the relative expression level of phosphorylated AKT (p-AKT) protein in the SAPCD2 overexpression group was higher than that in the overexpression control group, while that in the SAPCD2 knockdown group was lower than that in the knockdown control group, and the differences were statistically significant (all P < 0.01). Conclusions:SAPCD2 is highly expressed at both the transcriptional level and the protein level in ESCC tissues. SAPCD2 promotes the proliferation and migration of ESCC cells in vitro, which may be related to the AKT signaling pathway.

Objective:To analyze the mutation characteristics of rpoB gene in rifampicin-resistant Brucella strains. Methods:DNA of 4 rifampicin-resistant Brucella strains (JSY-26, G-9, WSY-13 and AW-3) isolated from Xinjiang Uygur Autonomous Region was selected, rifampicin rpoB gene was amplified by PCR and its nucleotide sequence was sequenced. The rpoB gene sequences of rifampicin-resistant Brucella standard strain (RB51) and sensitive strain (ALT-8) were used as reference, the mutation sites and types of the rpoB gene inside and outside the rifampicin resistance determination region (RRDR) of the 4 rifampicin-resistant Brucella strains were analyzed by Mega 7.0 software. Results:Through sequence alignment, both JSY-26 and WSY-13 strains underwent a single base point mutation at the RRDR 1 576 bp of the rpoB gene, with the base changing from guanine (G) to adenine (A). The G-9 strain underwent a single base point mutation at the RRDR 1 606 bp of the rpoB gene, with the base changing from cytosine (C) to A. The AW-3 strain showed 5 mutations of 3 types outside rpoB gene RRDR at 2 536, 2 537, 2 626, 2 636 and 2 654 bp, namely 3 insertion mutations [thymine (T) insertion once and C insertion twice], 1 deletion mutation (C deletion), and 1 single base point mutation (from G to C mutation).Conclusion:The RRDR mutations in the rpoB gene of the rifampicin-resistant Brucella strains are mainly characterized by single base point mutations, while multiple insertion and deletion mutations occur outside the RRDR.

Objective:To investigate the epidemic status and molecular characteristics of Borrelia burgdorferi in ticks in Xinjiang Uygur Autonomous Region (referred to as Xinjiang). Methods:From April to June 2020, 312 samples of Ixodes were collected in 6 areas of Yili, Alashankou, Hutubi, Qinghe, Fuhai and Wujiaqu, Xinjiang. Nested PCR and fluorescence quantitative PCR were used to detect Borrelia burgdorferi in ticks. The positive samples by both methods were genotyped and identified by nested PCR products. Results:The positive rates of nested PCR and fluorescence quantitative PCR were 8.97% (28/312) and 11.86% (37/312), respectively. Among them, the fluorescence quantitative PCR positive rate of Qinghe was the highest of 35.29% (12/34), and the positive rate of Fuhai was the lowest of 2.00% (1/50). The positive samples by both methods was 26. Genotyping results showed that 12 samples were highly homologous to Borrelia garinii, 10 copies to Borrelia burgdorferi sensu stricto, and 4 copies to Borrelia afzelii. Conclusions:The positive rate of Borrelia burgdorferi in ticks in Xinjiang is higher, which has confirmed that there are 3 pathogenic Borrelia burgdorferi genotypes in Xinjiang. The dominant genotype is Borrelia garinii, followed by Borrelia burgdorgferi sensu stricto genotype and Borrelia afzelii genotype.

Objective To identify molecular typing of Brucella abortus isolates in Xinjiang,and determine the identification ability of multiple locus variable-number tandem repeat analysis (MLVA).Methods The optimized Brucella AMOS-PCR was used for identification of Brucella (n =7) genus and species in Xinjiang from 2010-2015,and MLVA-16 was used to further identify the isolates.Results were compared with the data of the Brucella standard strain provided by the http://mlva.u-psud.fr database.Cluster analysis was carried out with Bionumerics 6.6.Results The results of AMOS-PCR and MLVA-16 were identical,all were Brucella abortus.Further classification results of the MLVA-16 showed that the strain in Xinjiang was type 3 of Brucella abortus,which was basically the same as that of the domestic Brucella.Conclusions The molecular typing of isolates separated in Xinjiang is type 3 of Brucella abortus.MLVA can identify Brucella at the level of species,and highly sensitive to Brucella biotype and isolates differences,which provides a basis for the traceability and evolution of brucellosis epidemic strains.

Objective To explore the drug sensibility of Brucella from bovine and sheep in Xinjiang.Methods Using paper diffusion method,19 drugs of 8 kinds of antibiotics including aminoglycosides,macrolides,sulfonamides,tetracyclines,β-lactams,fluoroquinolones,chloramphenicols and rifamycins,were tested.Drug sensitivity test was conducted on 57 Brucella strains isolated from bovine and sheep in Xinjiang from 2010 to 2016.Results The 57 Brucella strains were highly sensitive to doxycycline,tetracycline,streptomycin,tobramycin,gentamicin,amikacin,amoxicillin,ofloxacin,fleroxacin,ciprofloxacin and chloramphenicol,with the sensitivity rates were all higher than 90％;and they were highly resistance to azithromycin,clarithromycin and bactrim,with the drug resistance rates were all higher than 80％.Conclusion Brucella from bovine and sheep in Xinjiang is sensitive to tetracyclines,aminoglycosides,β-1actams,fluoroquinolones and chloramphenicols.

Objective To investigate the clinical and epidemiological characteristics of osteosarcoma in China and analyze the incidence and clinical treatment of osteosarcoma.Methods The clinical data of 1 593 patients with osteosarcoma in 7 bone cancer treatment centers from January 2000 to February 2017 were analyzed.We retrieved large samples of documents in the database,extracted the relevant data and compared the data with this study.SPSS 13 software was used for statistical analysis and each factor was tested by x2.Results Of 1 593 patients,984 were males and 609 were females.The ratio of male to female was 1.62∶ 1.The average age was 23.2 years (range from 3-80 years).The peak age of onset was 11-20 years (52.4％).There were 217 elderly patients (＞ 40 years old).The tumors occurred in 1 524 limbs and 69 in axial bone.The most common sites of disease were distal femur 706 cases,proximal tibia 375 cases,distal humerus 117 cases and others 395 cases.There were 1 154 cases (71％) around the knee joint.The axial bone included 18 cases of spine,49 cases of pelvis and 2 cases of rib.Preoperative biopsy was performed in 1 111 cases and incisional biopsy in 280 cases.Preoperative diagnosis could be made in 1 345 cases (84.4％ of all patients,accounting for 96.7％ of biopsy patients).There were 79 cases with metastasis at first visit,accounting for 5％.Preoperative chemotherapy was performed in 1 185 cases (74.4％).With the DIA preoperative chemotherapy 271 cases,DIA+MTX 251 cases,AP 149 cases.220 cases of tumor cell necrosis rate was evaluated after operationaccording to the Huvos classification.There were78 cases of grade Ⅰ,105 cases of Ⅱ grade,35 cases of Ⅲ and 2 cases of grade Ⅳ.There were 1 299 cases undergoingpostoperative chemotherapy (81.5％),1 306 patients undergoinglimb salvage surgery (82％).Thespecific operation with prosthetic replacement is the most common (911 cases,69.8％).The postoperative chemotherapy included DIA+MTX regimen 471 cases,DIA regimen 266 cases and AP regimen 98 cases.Before and after the operation,379 cases were treated with the same chemotherapy regimen and 666 cases were changed.A total of 18 large sample documents were retrieved in Chinese data base (5 684 cases).The sex ratio,age range,peak incidence and location of the disease were similar to those of this study.The average age was 1-2 years old younger.The percentage of lung metastases associated with initial visits washigher.The rate of preoperative and postoperative chemotherapy and limb salvage waslowand the rate of tumor necrosis wasbetter after chemotherapy.A total of 22 large sample literatures were searched in foreign database (12 850 cases).The ratio of men and women is 1.30∶1 and the proportion of women is higher than the domestic data.The average age was 1-2 years older.The ratio of the knee joint was lower.The percentage of lung metastases associated with initial visits washigher.The rate of preoperative and postoperative chemotherapy and limb salvage were similar to that of this study and the rate of tumor necrosis was better after chemotherapy.Conclusion The general situation of the incidence of osteosarcoma (sex,age,location of the disease) is not significantly different from the previous reports both at home and abroad.In the treatment,preoperative and postoperative chemotherapy rate and limb salvage surgery rate have improved significantly compared with the past.The chemotherapy program from the coexistence of various programs,gradually to the DIA+MTX and DIA program as the mainstream program.

Objective To isolate and extract effective antiendotoxin materials from Radix Paeoniae Rubra.Methods By the biosensor technology,effective antiendotoxin materials were isolated and extracted with general separation technology for Chinese traditional medicine.By ELISA mensuration for LPS and inhibition of TNF-? release from endotoxin-stimulated cells in vitro,the antiendotoxin activity of Radix Paeoniae Rubra was elucidated.Results The materials extracted had high binding capability to LPS and inhibitory effect on TNF-? release from endotoxin-stimulated cells in vitro.Conclusion Isolated and extracted effective antiendotoxin materials targeting LPS by the biosensor technology were valuable in searching antiendotoxin agents from Radix Paeoniae Rubra.The ability of Radix Paeoniae Rubra to neutralize LPS was assayed and proved very high.