An improved method for determination of the total oxalate in spinach was proposed. Oxalate in the spinach was precipitated at alkaline in CaC2O4 form. The precipitate was obtained by centrifugation and then acidified by HCIO4. KMnO4 was added and absorptivity was determined at 525 nm at a fixed time. This method was accurate and simple. Organic impurities existed in spinach could be removed by ultrviolet ray. The range of determination was 2?g/ml-35?g/ml. The recovery rate was 93-105% and the coefficient of variation was less than 5.35%.

Objective:This study aimed to investigate the expressions of EZH2 and Ki-67 in the salivary adenoid cystic carcino-ma (SACC) of humans and their correlation. Methods:A total of 42 cases of SACC tumor tissues and 5 cases of normal tissues were considered to determine the expressions of EZH2 and Ki-67 by immunohistochemistry. The relationship and correlation of such expres-sions with the clinicopathological characteristics were also analyzed. Results:The expression of EZH2 was notably higher in SACC than in normal tissues (P<0.05). EZH2 expression was detected in 66.67%(28/42) of the tumor tissues. This expression was correlated with pathological grade and clinical stage. By contrast, EZH2 expression did not correlate with gender, age, and localization. EZH2 was not expressed in normal tissues. The incidence of EZH2 expression in the Ki-67 positive group was 75.76%(25/33) and the incidence in the Ki-67 negative group was 33.33%(3/9). The difference between the two groups was statistically significant (P<0.05). Conclu-sion:The increased expression of EZH2 in SACC was related to tumor proliferation. EZH2 may participate in tumor cell proliferation via cell cycle management.

Objective To investigate the effect of X-ray irradiation on the expression of free ubiquitin in the serum, small intestine, and heart tissues of C57 BL/6N mice. Methods The amount of free ubiquitin protein in the serum and tissue homogenates was analyzed quantitatively with ELISA and Western Blotting assay. The mRNA expressions of free ubiquitin in the tissues were detected by RT-PCR. Results At 24 or 48 h after radiation, the free ubiquitin level in the serum and small intestine tissue increased asymptotically with increasing of radiation dose (F=183?1, 435?3, P <0?01). After 5 and 10 Gy X-ray irradiation, the concentration of serum free ubiquitin increased asymptotically with the extended response time (F=131?4, 442?9, P<0?01). Compared with the normal control group, at 24 h after 10 Gy X-ray irradiation, the expressions of free ubiquitin protein and mRNA in small intestine tissue were significantly up-regulated (t= -18?7, -10?1, P<0?01). However, there was no difference in the expressions of free ubiquitin protein and mRNA in the heart tissues between irradiated and nonirradiated groups (t = -2?0, 3?1, P >0?05). Conclusions Because of the high expressions of free ubiquitin protein in the radiosensitive mice tissues, X-ray radiation could increase the concentration of free ubiquitin in serum. The changes of free ubiquitin may be related to cellular radiosensitivity and tissue injury.

OBJECTIVE: To explore the association between single nucleotide polymorphism (SNPs) of KLK1 gene and cerebral hemorrhage in Changsha Han population. METHODS: We enrolled 273 patients with cerebral hemorrhage and 140 normal people. The SNPs (including rs3212855 and rs5515) of KLK1 gene were analyzed by Snapshot method and direct sequencing. RESULTS: We found rs5515 was not a polymorphic site in Changsha Han population. Genotype and allele frequency in rs3212855 were not different between patients with cerebral hemorrhage and the controls (P>0.05). The blood pressure level was not different between the genotype subgroups. CONCLUSION Neither rs5515 nor rs3212855 is associated with cerebral hemorrhage.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Cerebral Hemorrhage ; genetics ; China ; ethnology ; Female ; Genotype ; Humans ; Kallikreins ; genetics ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; genetics ; Young Adult

Objective:To investigate the radiosensitizing effect and underlying mechanism of STING agonist (c-di-AMP) on cutaneous melanoma cells.Methods:Human cutaneous melanoma cells (A375) were divided into four groups: the control group, 10 μmol/L c-di-AMP group, X-ray irradiation group and X-ray irradiation combined with c-di-AMP group. The radiosensitizing effect of c-di-AMP on A375 cells was detected by CCK-8-based viability assay, lactate dehydrogenase (LDH) release assay, flow cytometry-based apoptosis assay, and colony formation assay. Western blot analysis was used to determine the expressions of cell death-related proteins.Results:In combination with 10 Gy X-ray irradiation, 10 μmol/L c-di-AMP showed significant radiosensitization effect in A375 cells, which was evidenced by decreased cell activity ( t=5.11, P<0.05), increased cytotoxicity ( t=10.15, P<0.05) and cell apoptosis ( t=4.41, P<0.05) and reduced clone viability( t=6.30, 3.55, 5.45, 3.55, P<0.05). The calculated radiosensitization ratio of c-di-AMP to A375 cells was 1.88. Moreover, 10 μmol/L c-di-AMP further increased the expressions of cell death-related proteins induced by radiation in A375 cells. Conclusions:The STING agonist c-di-AMP can be used as a radiosensitizer for cutaneous melanoma, which may provide a novel strategy for radiotherapy.

In order to investigate the effects of LAL on the lipolysis and lipid synthesis of dairy adi-pocytes,the protein expressions of lipid synthesis-related molecules,acetyl-CoA carboxylase-1(ACC1),phosphorylated transcription factor-α(CEBPα),diacylglyceryl acyltransferase 2(DGAT2),sterol regulatory element binding protein 1(SREBP1),and peroxisome proliferator-ac-tivated receptor-γ(PPARγ),and lipolysis-related molecules,lipid droplet coated protein-1(PLIN1),triglyceride lipase(ATGL),hormone-sensitive lipase(HSL),p-HSL,and the target pro-tein lysosomal acid lipase(LAL),were detected in adipose tissues of healthy dairy cows and ketosis cows.The lipid synthesis and lipolysis-related protein expressions in adipocytes were detected by Western blot technology.The primary bovine adipocytes were cultured in vitro with overexpressed LAL,and the lipolysis model of adipocytes was constructed by adding isoproterenol(ISO).The re-sults showed that the expression of LAL in adipose tissue of ketosis cows was significantly lower than that of healthy cows(P＜0.01).Compared with healthy cows,the protein expression levels of lipid synthesis-related proteins ACC1,CEBPα,DGAT2,SREBP1 and PPARγin adipose tissue of clinical ketosis cows were significantly decreased,while the protein expression and phosphorylation levels of lipolysis-related proteins,PLIN1,ATGL,and p-HSL were significantly increased.The a-bove results confirmed that the lipid synthesis of adipose tissue of ketosis cows was inhibited,and the lipolysis was enhanced.In vitro results showed that ISO could downregulate the protein ex-pression levels of lipid synthesis related molecules,and upregulate the protein expression and phosphorylation levels of lipid lysis related molecules in bovine adipocytes.The content of basal lipid synthesis and ISO-induced lipid synthesis proteins in bovine adipocytes of LAL overexpres-sion group was significantly increased,while the content of basal lipid lysis and ISO-induced lipid lysis proteins was significantly decreased.In conclusion,in vivo and in vitro studies have shown that LAL can inhibit the lipid lysis of bovine adipocytes and promote the lipid synthesis of bovine adipocytes.

Objective:To investigate the impacts of ionizing radiation on protein expression profiles in esophageal tissues of rats using quantitative proteomics, in order to reveal the molecular mechanisms underlying the onset and development of radiation-induced esophagitis (RIE).Methods:A total of twenty-four male SD rats were divided by simple randomization into three groups: the control, 25 Gy irradiation, and 35 Gy irradiation groups, and their esophageal tissues were collected at 7 d post-irradiation to extract total protein. Then, changes in the protein expression profiles of the esophageal tissues in irradiated rats were investigated using tandem mass tag (TMT)-labeled quantitative proteomics and bioinformatics analysis. Additionally, the expressions of two key proteins, Hp and Ndufs4, were validated using immunohistochemistry and Western blot.Results:A comparison with the control group revealed a total of 847 differentially expressed proteins (DEPs; 483 up-regulated and 364 down-regulated) following 25 Gy irradiation and 699 DEPs (443 up-regulated and 256 down-regulated) following 35 Gy irradiation. Different radiation doses led to common 326 up-regulated proteins, which were mainly involved in biological processes and signaling pathways related to immune and inflammatory responses, and 210 down-regulated proteins, which were primarily involved in biological processes and signaling pathways related to energy production and metabolism. Furthermore, a total of 155 proteins were screened using a constructed protein protein interaction(PPI) network. Of these proteins, the up-regulated ones were most associated with three functional pathways, namely innate immune responses, complement and coagulation cascades, and innate immune system, while the down-regulated ones were most associated with energy acquisition via oxidizing organic compounds, oxidative phosphorylation, and the tricarboxylic acid (TCA) cycle and respiratory electron transfer. These functions were enriched with nine complement-related up-regulated and five mitochondria-related down-regulated proteins, respectively. Ionizing radiation significantly up-regulated Hp ( t = 27.94, 10.96, P<0.001) and down-regulated Ndufs4 ( t = 59.27, 54.07, P<0.001), consistent with the protein sequencing result. Conclusions:Ionizing radiation can change the protein expression profiles in the esophageal tissues of rats, and these DEPs are involved in multiple radiobiology-related functional pathways such as immune processes, inflammatory responses, and abnormal energy metabolism. Screening and validation of key proteins are helpful for identifying potential biomarkers of radiation-induced esophagitis.

Threatened abortion is a common disease of obstetrics and gynecology and one of the diseases responding specifically to traditional Chinese medicine （TCM）. The China Association of Chinese Medicine organized experts in TCM obstetrics and gynecology， Western medicine obstetrics and gynecology， and pharmacology to deeply discuss the advantages of TCM and integrated Chinese and Western medicine treatment as well as the medication plans for threatened abortion. After discussion， the experts concluded that chromosome， endocrine， and immune abnormalities were the key factors for the occurrence of threatened abortion， and the Qi and blood disorders in thoroughfare and conception vessels were the core pathogenesis. In the treatment of threatened abortion， TCM has advantages in preventing miscarriages， alleviating clinical symptoms and TCM syndromes， relieving anxiety， regulating reproductive endocrine and immune abnormalities， personalized and diversified treatment， enhancing efficiency and reducing toxicity， and preventing the disease before occurrence. The difficulty in diagnosis and treatment of threatened abortion with traditional Chinese and Western medicine lies in identifying the predictors of abortion caused by maternal factors and the treatment of thrombophilia. Recurrent abortion is the breakthrough point of treatment with integrated traditional Chinese and Western medicine. It is urgent to carry out high-quality evidence-based medicine research in the future to improve the modern diagnosis and treatment of threatened abortion with TCM.