Overexpression SPRY2 Suppresses Esophageal Squamous Cell Carcinoma Progression Via Inactivation of ERK/AKT Signaling / La Sobreexpresión SPRY2 Suprime la Evolución del Carcinoma de Células Escamosas de Esófago Mediante la Inactivación de la Señalización ERK/AKT

SUMMARY: Esophageal cancer is one of the most aggressive gastrointestinal cancers. Invasion and metastasis are the main causes of poor prognosis of esophageal cancer. SPRY2 has been reported to exert promoting effects in human cancers, which controls signal pathways including PI3K/AKT and MAPKs. However, the expression of SPRY2 in esophageal squamous cell carcinoma (ESCC) and its underlying mechanism remain unclear. In the present study, we aimed to investigate the detailed role of SPRY2 in the regulation of cell proliferation, invasion and ERK/AKT signaling pathway in ESCC. It was identified that the expression level of SPRY2 in ESCC was remarkably decreased compared with normal tissues, and it was related to clinicopathologic features and prognosis ESCC patients. The upregulation of SPRY2 expression notably inhibited the proliferation, migration and invasion of Eca-109 cells. In addition, the activity of ERK /AKT signaling was also suppressed by the SPRY2 upregulation in Eca-109 cells. Our study suggests that overexpression of SPRY2 suppress cancer cell proliferation and invasion of by through suppression of the ERK/AKT signaling pathways in ESCC. Therefore, SPRY2 may be a promising prognostic marker and therapeutic target for ESCC.

El cáncer de esófago es uno de los cánceres gastrointestinales más agresivos. La invasión y la metástasis son las principales causas de mal pronóstico del cáncer de esófago. Se ha informado que SPRY2 ejerce efectos promotores en los cánceres humanos, que controla las vías de señales, incluidas PI3K/AKT y MAPK. Sin embargo, la expresión de SPRY2 en el carcinoma de células escamosas de esófago (ESCC) y su mecanismo subyacente aún no están claros. En el presente estudio, nuestro objetivo fue investigar el papel detallado de SPRY2 en la regulación de la proliferación celular, la invasión y la vía de señalización ERK/AKT en ESCC. Se identificó que el nivel de expresión de SPRY2 en ESCC estaba notablemente disminuido en comparación con los tejidos normales, y estaba relacionado con las características clínico-patológicas y el pronóstico de los pacientes con ESCC. La regulación positiva de la expresión de SPRY2 inhibió notablemente la proliferación, migración e invasión de células Eca-109. Además, la actividad de la señalización de ERK/AKT también fue suprimida por la regulación positiva de SPRY2 en las células Eca-109. Nuestro estudio sugiere que la sobreexpresión de SPRY2 suprime la proliferación y la invasión de células cancerosas mediante la supresión de las vías de señalización ERK/AKT en ESCC. Por lo tanto, SPRY2 puede ser un marcador de pronóstico prometedor y un objetivo terapéutico para la ESCC.

KDM4A promotes the migration and invasion of breast cancer cell line MDA-MB-231 by downregulating BMP9 / 中国癌症杂志

Background and purpose:Exogenous bone morphogenetic protein 9(BMP9)inhibits the malignant progression of human breast cancer,but its expression is often abnormally low in breast cancer.In this study,we intended to explore the expression and role of epigenetically-modified histone lysine-specific demethylase 4A(KDM4A)in breast cancer,and to investigate the relationship between KDM4A and BMP9 and its possible regulatory mechanism.Methods:The expression of KDM4A in breast cancer and its relationship with BMP9 were analyzed by bioinformatics and verified by real-time fluorescence quantitative polymerase chain reaction(RTFQ-PCR)and Western blot.Chromatin immunoprecipitation(ChIP)verified the regulatory role of KDM4A on BMP9,and RNA stability experiments and CHX protein stability experiments verified the effect of KDM4A in BMP9 expression.Exogenous recombinant MDA-MB-231 cells transfected with KDM4A small interfering RNA(siKDM4A)or infected with siBMP9 adenovirus(Ad-siBMP9)were constructed using RNA interference technology and adenoviruses knocking down BMP9,and the migratory and invasive abilities of the cells were detected by scratch healing assay and transwell assay,respectively.Results:Bioinformatics analysis showed that the expression of KDM4A was significantly higher in breast cancer than in normal tissues,and there was a negative correlation between the expression of KDM4A and that of BMP9 in breast cancer;RTFQ-PCR and Western blot showed that KDM4A was highly expressed in different breast cancer cell lines,and the knockdown of KDM4A significantly up-regulated BMP9.ChIP experiment confirmed that KDM4A could be significantly enriched in the promoter region of BMP9 gene,reducing its histone lysine 36 position instead of position 4 methyl status,thus silencing the expression of BMP9.RNA stability assay and CHX protein stability assay confirmed that KDM4A had no significant effect on the mRNA of BMP9,but could affect its protein degradation.After knocking down KDM4A,the migration and invasion abilities of breast cancer cells MDA-MB-231 were significantly inhibited,and this effect could be partially reversed by knocking down BMP9.Conclusion:KDM4A is highly expressed in breast cancer and breast cancer cell MDA-MB-231,and can silence its expression by down-regulating the level of histone methylation in the promoter region of the BMP9 gene,as well as affecting the stability of BMP9 at the protein level rather than at the level of mRNA,and promoting the migration and invasion of breast cancer.

Current status of global colorectal cancer prevalence, prevention and control / 中华肿瘤杂志

Objective: This paper provides a brief overview of the epidemiology of colorectal cancer in China and around the world, and discusses how to prevent colorectal cancer to reduce its disease burden. Method: Using the official database of GLOBOCAN 2020, the China Cancer Registry Annual Report compiled by the National Cancer Center, and data from CONCORD-3.Data management was performed by Microsoft Excel 2016 and R 4.2.1 Relevant graphs were generated using the ggplot2 package for result visualization. Result: An estimated 1 931 590 people were diagnosed with colorectal cancer worldwide in 2020 with an age-standardized incidence rate of 19.5 per 100 000. There were about 935 173 deaths caused by colorectal cancer internationally, with an age-standardized mortality rate of 9.0 per 100 000. Overall, colorectal cancer was the fourth most commonly diagnosed cancer and the third leading cause of cancer-related death worldwide in 2020. In China, the age-standardized incidence rate and mortality rate of colorectal cancer was 17.3 per 100 000 and 7.8 per 100 000, respectively. Gender differences in trends were observed, with a decreasing trend in incidence and mortality among females and an increasing trend in incidence and mortality among males. The primary risk factors for colorectal cancer include age, genetic factors, gastrointestinal disorders, dietary habits, and lifestyle et al. Conclusions: Colorectal cancer poses a significant burden globally and in China. The occurrence of colorectal cancer is closely related to physiology, genetics, behavioral habits, lifestyle, and disease factors. To better control the colorectal cancer burden with the lowest cost, specific measures should be taken to reduce exposure to established risk factors. By combining the disease prevention and control strategies of tertiary prevention in China with the characteristic factors of colorectal cancer, the incidence and mortality of colorectal cancer may be effectively controlled.

Current status of global colorectal cancer prevalence, prevention and control / 中华肿瘤杂志

Objective: This paper provides a brief overview of the epidemiology of colorectal cancer in China and around the world, and discusses how to prevent colorectal cancer to reduce its disease burden. Method: Using the official database of GLOBOCAN 2020, the China Cancer Registry Annual Report compiled by the National Cancer Center, and data from CONCORD-3.Data management was performed by Microsoft Excel 2016 and R 4.2.1 Relevant graphs were generated using the ggplot2 package for result visualization. Result: An estimated 1 931 590 people were diagnosed with colorectal cancer worldwide in 2020 with an age-standardized incidence rate of 19.5 per 100 000. There were about 935 173 deaths caused by colorectal cancer internationally, with an age-standardized mortality rate of 9.0 per 100 000. Overall, colorectal cancer was the fourth most commonly diagnosed cancer and the third leading cause of cancer-related death worldwide in 2020. In China, the age-standardized incidence rate and mortality rate of colorectal cancer was 17.3 per 100 000 and 7.8 per 100 000, respectively. Gender differences in trends were observed, with a decreasing trend in incidence and mortality among females and an increasing trend in incidence and mortality among males. The primary risk factors for colorectal cancer include age, genetic factors, gastrointestinal disorders, dietary habits, and lifestyle et al. Conclusions: Colorectal cancer poses a significant burden globally and in China. The occurrence of colorectal cancer is closely related to physiology, genetics, behavioral habits, lifestyle, and disease factors. To better control the colorectal cancer burden with the lowest cost, specific measures should be taken to reduce exposure to established risk factors. By combining the disease prevention and control strategies of tertiary prevention in China with the characteristic factors of colorectal cancer, the incidence and mortality of colorectal cancer may be effectively controlled.

Application effect of discharge preparation service based on theory of goal attainment on patients with cervical spinal cord injury / 中华创伤杂志

Objective:To investigate the application effect of discharge preparation service based on theory of goal attainment on patients with cervical spinal cord injury.Methods:A retrospective cohort study was conducted to analyze the clinic data of 60 patients with cervical spinal cord injury admitted to Zhengzhou Orthopedics Hospital from January 2017 to December 2022, including 49 males and 11 females, aged 23-79 years [(52.2±13.5)years]. Patients were all treated with cervical decompression fusion and internal fixation. Patients admitted from January 2017 to December 2019 were treated with conventional nursing intervention (conventional nursing group, n=30) and patients admitted from January 2020 to December 2022 were treated with discharge preparation service based on theory of goal attainment (discharge preparation service group, n=30). The readiness for hospital discharge of the two groups was compared using the Chinese version of Readiness for Hospital Discharge Scale (RHDS) at 4 hours before discharge. The degree of cervical spinal cord dysfunction of the two groups were compared using Japanese Orthopedic Association (JOA) score before intervention, at discharge and at 6 months after discharge. The complication and unplanned readmission rates of the two groups were compared at 6 months after discharge. Results:All the patients were followed up for 6 months. At 4 hours before discharge, the scores of the three parameters of RHDS containing personal status, adaptability and anticipatory support and the total score of the discharge preparation service group were (20.9±3.5)points, (35.9±2.2)points, (30.4±3.0)points and (87.1±7.8)points respectively, higher than those of the conventional nursing group [(16.2±1.7)points, (32.5±2.2)points, (26.3±2.1)points and (75.0±5.6)points respectively] ( P<0.01). There was no statistically significant difference in the JOA score of the two groups before intervention ( P>0.05). The JOA scores of the discharge preparation service group at discharge and at 6 months after discharge were (11.8±1.7)points and (13.8±1.5)points respectively, higher than those of the conventional nursing group [(10.3±1.8)points and (11.6±1.9)points respectively] ( P<0.01). At 6 months after discharge, the complication rate of the discharge preparation service group was 6.7% (2/30), lower than that of the conventional nursing group [36.7% (11/30)] ( P<0.05). The unplanned readmission rate of the discharge preparation service group was 3.3% (1/30), lower than that of the conventional nursing group [23.3% (7/30)] ( P<0.05). Conclusion:For patients with cervical spinal cord injury, discharge preparation service based on theory of goal attainment can improve the discharge readiness, promote spinal functional recovery and reduce the complication and unplanned readmission rates.

Formate dehydrogenase and its application in biomanufacturing of chiral chemicals / 生物工程学报

The redox biosynthesis system has important applications in green biomanufacturing of chiral compounds. Formate dehydrogenase (FDH) catalyzes the oxidation of formate into carbon dioxide, which is associated with the reduction of NAD(P)+ into NAD(P)H. Due to this property, FDH is used as a crucial enzyme in the redox biosynthesis system for cofactor regeneration. Nevertheless, the application of natural FDH in industrial production is hampered by low catalytic efficiency, poor stability, and inefficient coenzyme utilization. This review summarized the structural characteristics and catalytic mechanism of FDH, as well as the advances in protein engineering of FDHs toward improved enzyme activity, catalytic efficiency, stability and coenzyme preference. The applications of using FDH as a coenzyme regeneration system for green biomanufacturing of chiral compounds were summarized.

Changing trend of incidence and mortality of esophageal cancer during 2010-2016 in Henan Province, China / 中华肿瘤杂志

Objective: To estimate the incidence and mortality of esophageal cancer in 2016 and their changing trend during 2010-2016 according to the cancer registration data in Henan province. Methods: The data quality including completeness, validity, and reliability of local registries which submitted the cancer registration data of 2016 were assessed according to the criteria of Guideline on Cancer Registration in China and IARC/IACR. Esophageal cancer cases (ICD10: C15) were extracted from the database, and the incidence and mortality stratified by gender, age, and areas (urban/rural) were calculated, the incidence and mortality of provincial cancer were estimated combined with provincial population data. China's 2000 census population and Segi's population were used to calculate the age-standardized rate. Joinpoint model was used to estimate the changing trend of age standardized incidence and mortality along with the calendar year. Results: Approximately 40.10 thousand new esophageal cancer cases were diagnosed in Henan in 2016, accounting for 13.46% of all new cancer cases, and it ranked the third among cancer of all sites. The crude incidence of esophageal cancer was 37.21/100 000 with an age-standardized incidence rate by China standard population (ASIRC) of 26.74/100 000 and an age-standardized incidence rate by world standard population (ASIRW) of 27.12/100 000. The incidence of esophageal cancer in males was higher than that in females, with the ASIRC of 34.53/100 000 and 19.19/100 000, respectively. It was higher in rural areas than that in urban areas, with the ASIRC of 28.13/100 000 and 20.90/100 000, respectively. About 29.30 thousand deaths of esophageal cancer in Henan in 2016, accounting for 15.61% of all cancer deaths in Henan, which ranked the third among cancer of all sites. The crude mortality rate was 27.14/100 000 with an age-standardized mortality rate by China standard population (ASMRC) of 18.74/100 000 and an age-standardized mortality rate by world standard population (ASMRW) of 18.78/100 000. The mortality in males was higher than that in females, with the ASMRC of 24.78/100 000 and 13.12/100 000, respectively. It was also higher in rural areas than that in urban areas, with the ASMRC of 19.48/100 000 and 15.73/100 000, respectively. The ASIRC and ASMRC were declining with annual percent change (APC) of 3.12% (APC=-3.12%; 95%CI: -5.30%, -0.90%; P=0.015) and 2.47% (APC=-2.47%; 95%CI: -4.70%, -0.20%; P=0.039) during 2010-2016. However, the significant declining trend was only observed in rural areas in Henan, and the changing trend was same between males and females. Conclusions: The incidence and mortality of esophageal cancer are declining since 2010, however, the disease burden remains large in Henan. Therefore, comprehensive prevention and control efforts should be strengthened according to its epidemic characteristics and risk factors.

Changing trend of incidence and mortality of stomach cancer during 2010-2016 in Henan Province, China / 中华肿瘤杂志

Objective: To estimate stomach cancer incidence and mortality in Henan, 2016 and analyze the trend of stomach cancer incidence and mortality from 2010 to 2016. Methods: Stomach cancer related data in 2016 was extracted from Henan cancer registration and follow-up system. All data were qualified in validity, reliability and completeness according to the Guideline on Cancer Registration in China and International Agency for Research on Cancer (IARC/IACR). The incidence and mortality of stomach cancer were estimated by areas, gender and age based on the quality data and the registered population data of Henan province in 2016. The epidemic trend of stomach cancer was also been evaluated based on the age-standardized incidence and mortality by Chinese population (ASR China) from 2010 to 2016. Results: In 2016, the estimated incident cases of stomach cancer were 44 311. The incidence was 41.07/100 000, ASR China was 30.17/100 000, ASR by world population (ASR world) was 30.36/100 000, and the cumulative incidence rate was 3.84%. The incidences of male and female were 55.65/100 000 and 25.35/100 000, respectively. Meanwhile, 32 927 people died of stomach cancer in Henan. The mortality was 30.52/100 000, ASR China was 21.45/100 000, ASR world was 21.54/100 000, and the cumulative mortality was 2.53%. From 2010 to 2016, both the ASR China for incidence and mortality of stomach cancer in Henan showed a steady downward trend. In rural, the ASR China for incidence and mortality decreased rapidly, while the stable trend was observed in urban. Nevertheless, the incidence and mortality of stomach cancer in rural were still higher than those in urban. Conclusions: The incidence and mortality of stomach cancer in Henan province showed steadily declining trend from 2010 to 2016, and the geographical distribution difference between rural and urban areas was gradually narrowing. However, the disease burden was still high in 2016.

Text Analysis of Enterprise Social Responsibility Report of Pharmaceutic al Industry in China ：Taking Listed Com- panies as an Example / 中国药房

OBJECTIVE：To provide reference for pharmaceutical enterprises to better fulfill their social responsibilities . METHODS：Taking the pharmaceutical enterprises listed in Shanghai and Shenzhen stock markets as the research objects ，the social responsibility texts and environmental protection texts of the included enterprises from 2017 to 2019 were classified and analyzed according to the three dimensions of time ，ownership and drug production type by using ROST Content Mining 6.0 software. The semantic network and node centrality were analyzed. RESULTS ：From the social responsibility texts of the included enterprises，it could be seen that from the perspective of time dimension ，the included enterprises paid more attention to “employees”，“social responsibility ”and“development”in 2017-2019，and the high-frequency words in the three years were relatively stable. From the perspective of ownership ，state-owned holding enterprises paid more attention to “development”and “targeted poverty reduction ”，while private holding enterprises paid more attention to “employees”，and collective holding enterprises paid more attention to “social responsibility ”. From the perspective of drug production type ，the word frequency of “targeted poverty reduction ”of TCM pharmaceutical enterprises was higher. From the environmental protection texts of the included enterprises，it could be seen that from the time dimension ，the included enterprises paid more and more attention to environmental protection in recent years ，and the description of environmental protection in the texts was more detailed. From the perspective of ownership，private holding enterprises paid more attention to environmental protection than collective holding enterprises and state-owned enterprises. From the perspective of drug production type ，due to the different types of drugs ，chemical drugs ，TCM， biological drugs and active pharmaceutical ingredients enterprises had their own characteristics in pollutants and their treatment processes. The results of semantic network and node centrality analysis showed that ，“development”，“protection” and “targeted poverty reduction ”were three high-frequency words with high centrality in the semantic analysis network of social responsibility text ，the middle centrality was 5.555，1.695，1.521 respectively；“emission”and“testing”were two key words with high centrality in the semantic analysis network of environmental protection texts ，the middle centrality was 3.856， 1.914 respectively，which further reflected that the included enterprises paid more attention to the above issues in terms of social responsibility performance and environmental protection. CONCLUSIONS ：The listed pharmaceutical enterprises have a broad consensus on social responsibility in China ，and have well fulfilled their social responsibility ，which is well combined with the current social hot spots. For environmental protection ，as a high pollution industry ，the pharmaceutical enterprises also performs the responsibility of environmental protection.

Development of a fast and sensitive method for direct analysis of insulin aspart in human plasma / 药学学报

A fast and sensitive UPLC-MS/MS method was established for the direct quantification of insulin aspart in human plasma. The plasma samples were extracted by solid phase extraction (SPE), an ESI ion source was used and operated in the positive ion mode with multiple reaction monitoring (MRM). Bovine insulin was chosen as internal standard and the chromatographic separation of insulin aspart was performed on Waters ACQUITY UPLC CSH C18 column (50 mm×2.1 mm, 1.7 μm). A mixture of acetic acid aqueous solution and acetonitrile with acetic acid at a flow rate of 0.6 mL·min-1 in gradient elution mode was employed as mobile phase. We found that the method was validated over the range of 0.200-10.0 ng·mL-1 for insulin aspart and showed excellent linearity. The intra-and inter-assay accuracy and precision were below 14.5% and the recovery was 36.7%-41.7% over the three concentration levels evaluated. The UPLC-MS/MS method was selective, accurate, sensitive and robust, and the method was successfully applied in supporting the pharmacokinetic research of two insulin aspart injections (Test Product and NovoRapid®) in heathy male subjects. This clinical trial was approved according to the Ethics Committee of West China Hospital, Sichuan University (2017 Clinical Trial (Western Medicine) Approval (148)).

The molecular mechanism of fibroblast growth factor 21-inhibited leptin expression in adipocytes / 生理学报

The present study was aimed to clarify the signaling molecular mechanism by which fibroblast growth factor 21 (FGF21) regulates leptin gene expression in adipocytes. Differentiated 3T3-F442A adipocytes were used as study object. The mRNA expression level of leptin was detected by fluorescence quantitative RT-PCR. The phosphorylation levels of proteins of signal transduction pathways were detected by Western blot. The results showed that FGF21 significantly down-regulated the mRNA expression level of leptin in adipocytes, and FGF21 receptor inhibitor BGJ-398 could completely block this effect. FGF21 up-regulated the phosphorylation levels of ERK1/2 and AMPK in adipocytes. Either ERK1/2 inhibitor SCH772984 or AMPK inhibitor Compound C could partially block the inhibitory effect of FGF21, and the combined application of these two inhibitors completely blocked the effect of FGF21. Neither PI3K inhibitor LY294002 nor Akt inhibitor AZD5363 affected the inhibitory effect of FGF21 on leptin gene expression. These results suggest that FGF21 may inhibit leptin gene expression by activating ERK1/2 and AMPK signaling pathways in adipocytes.

Chemical profiling from water extract of Wikstroemia indica by UPLC-Q-TOF-MS/MS / 中国中药杂志

In this study,a method using ultra performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry( UPLC-Q-TOF-MS/MS) was established to identify complicated chemical constituents of Wikstroemia indica. Chromatographic separation was performed on an AcclaimTMRSLC 120-C18 column( 2. 1 mm×100 mm,2. 2 μm) using gradient elution with 0. 2% ammonium formate buffer salt solution( A)-0. 2% ammonium formate buffer salt solution methanol( B) as mobile phase. The column temperature was maintained at 30 ℃. The analytes were determined by positive and negative ion modes with electro-spray ionization source. A total of 52 compounds( including eleven coumarins,thirteen flavonoids,ten lignans,two amides,four phenolic acids,six sesquiterpenes and six other compounds) were identified or tentatively characterized from the water extract of W. indica by comparing their retention times and MS spectra with those of authentic standards or literature datas. Three compounds were found for the first time from W.indica namely isomer of indicanone,β-hydroxypropiovanillone and epiprocurcumenol. Furthermore,the fragmentation rules of some compounds were speculated and summarized. In addition,the cleavage pathways of guaiane sesquiterpenes were described for the first time,which can provide reference for studying the fragmentation pathways of similar compounds. This study provides an easy way to identify chemical constituents of traditional Chinese medicine and a basis for the further study on chemical fundamentals of W. indica.

Facilitative glucose transporters: expression, distribution and the relationship to diseases / 生理学报

Facilitative glucose transporters (GLUT) are proteins that mediate glucose transmembrane transport in the form of facilitated diffusion, which play an important role in regulating cell energy metabolism. There are many breakthroughs in researches of facilitative GLUT in recent years. It has been known that there are 14 subtypes of facilitative GLUT with obvious tissue specificity in distribution and physiological function. In the present review, the tissue and cellular distribution, subcellular localization, expression regulation, physiological function and the relationship to diseases of facilitative GLUT subtypes were summarized, in order to further understand their physiological and pathophysiological significances.

The relationship between inflammatory factors and the risk of breast cancer in Kailuan cohort / 中华疾病控制杂志

Objective To investigate whether elevated baseline levels of high sensitivity C-Reactive Protein (hsCRP) and neutrophil (NE) are associated with an increased risk of breast cancer in Kailuan female cohort. Methods Females from Kailuan cohort (2006-2007) were included in this study. Information on check-up, hsCRP and NE were collected at baseline for all subjects. Multivariable Cox proportional hazards regression models were used to calculate hazard ratios (HR) and 95% confidence intervals (95%CI) of association between baseline hsCRP and NE values and breast cancer risk. Results By December 31, 2015, a total of 18 866 participants were enrolled in this study. During the follow-up, 183 new cases of breast cancer were observed. All participants were divided into three groups according to the level of hsCRP (3 mg/L). The cumulative incidence of breast cancer were 829/105, 1 211/105 and 1 495/105 in these 3 groups, respectively ( 2=12.08， P=0.002). Compared with participants with lower hsCRP levels (3 mg/L) levels had significantly increased risk of breast cancer (HR=1.71,95%CI: 1.18-2.47, P=0.005), howerver, we didn’t find the statistically significant association between NE level (0.05). Conclusions Elevated levels of hsCRP at baseline might increase the risk of breast cancer in females.

Sciatic nerve leachate of cattle causes neuronal differentiation of PC12 cells via ERK1/2 signaling pathway

Previous studies have shown that the sciatic nerve has neurotrophic activity, and nerve regeneration, differentiation, and axon outgrowth can be modulated by different sciatic nerve preparations. However, numerous animals may have to be sacrificed to obtain enough sciatic nerves to make a sciatic nerve preparation. Some studies have demonstrated that the role of sciatic nerve preparations in neural differentiation depends on the neurotrophins that Schwann cells secrete, and these factors are highly conserved among different species. To reduce the use of experimental animals, in this study, we made a leachate by using the sciatic nerve of cattle and explored its effect on neuronal differentiation of rat PC12 cells (a useful model for studying neuronal differentiation). Results showed the neurite outgrowth of PC12 cells treated with the cattle sciatic nerve leachate for 3, 6, and 9 days was significantly improved, and the expressions of β3-tubulin and microtubule-associated protein 2 (two neuron-specific proteins) were increased. Moreover, the ERK1/2 signaling pathway was activated after PC12 cells were incubated with cattle sciatic nerve leachate for 9 days. Thus, a sciatic nerve leachate obtained from cattle can effectively induce neuronal differentiation of rat PC12 cells via ERK1/2 signaling pathway.

Research Methods for Structural Analysis of Lactic Acid Bacteria Induced Exopolysaccharides / 分析化学

Exopolysaccharides (EPSs), produced by lactic acid bacteria (LAB), have been used primarily to improve the quality and taste of food, also possess a variety of unique biological functions, such as immunoregulation and anti-tumor activities. The diversity of molecular structural characteristics of LAB-generated EPSs represents one of the main factors responsible for this plethora of functions. Accordingly, the structural analysis of the EPSs produced by LAB is both a prerequisite and basis for examining its functional and structure-activity relationships. In this article, we summarized the current progress of key methodologies involved in the structural analysis of LAB-generated EPSs, including their isolation, purification, primary structure and advances in structural research. A comprehensive discussion regarding the application of chemical analysis, instrument analysis and computer aided technology in the structure analysis of LAB-generated EPSs was provided. Further, the future development of the research on the structure of LAB-generated EPSs was presented.

Correlation between gene polymorphism of cytotoxic T lymphocyte associated antigen-4 and Graves disease in Qinghai Tibetan / 中华地方病学杂志

Objective To investigate the correlation between gene polymorphism of cytotoxic T lymphocyte associated antigen-4 (CTLA-4) and Graves disease (GD) in Qinghai Tibetan.Methods Using retrospective analysis methods,totally 130 cases of GD were selected randomly from June 2012 to November 2016 in the People's Hospital of Qinghai Province;meanwhile,110 normal control cases were selected randomly from Qinghai Tibetan.Then the genotype and allele of CTLA-4 were detected by the method of polymerase chain reaction restriction fragment length polymorphism (RFLP-PCR).Results The distribution of CTLA-4 genotype frequencies (AA,AG,GG) was different between normal control cases and GD in Qinghai Tibetan [6.2％ (8/130) vs 26.4％ (29/110),50.0％ (65/130)vs 58.2％ (64/110),43.8％ (57/130) vs 15.4％ (17/110),x2 =32.105,P ＜ 0.05].Allele (A,G) frequencies were compared between GD and control,the differences were statistically significant [31.2％(81/260) vs 55.5％ (122/220),68.8％ (179/260) vs 44.5％ (98/220),x2 =28.834,P ＜ 0.05].Conclusion Polymorphisms of CTLA-4 exon 1 (49A/G)genotype and allele are closely correlated with GD in Qinghai Tibetan.

Enterovirus 71 inhibited interferon signaling by downregulating expression of IRF9 / 中国免疫学杂志

Objective:To explore the mechanism of EV71 antagonizing IFN signaling pathway.Methods: RD cells were infected or un-infected with EV71.Then the cells were treated with or without IFN-β.The four groups (the control group,the EV71 group,the IFN-β group,the EV71+IFN-β group) were detected by molecular biology techniques.The expression of interferon stimulated genes (ISGs) were detected by Real-time PCR,while the protein levels of STAT1 and IRF9 were examined by Western blot assay.By preparing the cytosolic and nuclear fractions,the translocation of p-STAT1 was monitored through Western blot assay.Results:Compared with the IFN-β group,the mRNA level of OAS1,MX1 and ISG54 in the EV71+IFN-β group was down regulated by 47%, 50% and 48%,respectively,indicating that EV71 inhibited the expression of ISGs.The results also showed that EV71 did not effect the protein level and phosphorylation of STAT1.Moreover,we found that p-STAT1 was translocated into neuclear in IFN-β group,while p-STAT1 was located in the cytoplasm in the EV71+IFN-β group.And the expression of IRF9 was boviously down regulated in EV71+IFN-β group compared with that in IFN-β group,suggesting that EV71 blocked the expression of IRF9 induced by IFN-β.Conclusion:EV71 inhibited the IFN signaling pathway by downregulating the expression of IRF9 induced by IFN-β.

Analysis of Traditional Chinese Medicine Constitution of Patients with Metabolic Syndrome / 广州中医药大学学报

Objective To investigate the characteristics of traditional Chinese medicine(TCM) constitution of patients with metabolic syndrome(MS), so as to provide individualized therapy for the treatment and rehabilitation of MS. Methods The data of a total of 328 MS patients were collected, covering body height, body mass,body mass index(BMI),waist circumference,blood pressure,blood glucose,blood lipids such as triglyceride(TG),total cholesterol(TC),low-density lipoprotein cholesterol(LDL-C),high-density lipoprotein cholesterol(HDL-C), etc.. TCM constitution types of the MS patients were classified according to Self-rating Scale for Traditional Chinese Medicine Constitution Classification and Discrimination. Results (1)In the 328 cases,the patients with increased TG or having treated with lipid-lowering therapy,the patients complicated with increased blood pressure or having treated with antihypertensive therapy,and the patients with the previous diagnosis of hypertension were dominant,accounted for 41 cases(12.50%).(2)Bias constitution was predominant, showing in 308 cases and accounting for 93.90%. Phlegm-damp constitution was mostly commonly seen in the bias constitution, accounted for 78 cases(23.78%). And then the damp-heat constitution, Qi stagnation constitution, complicated constitution, Qi deficiency constitution, yang deficiency constitution, blood-stasis constitution, yin deficiency constitution and specific constitution came next orderly. Twenty patients had the harmony constitution, accounting for 6.10%. (3)MS patients with phlegm-damp constitution had higher body mass and BMI than the patients with harmony constitution, the difference being significant (P < 0.05). The differences of TG were significant between the phlegm-damp constitution and the specific constitution(P <0.05). Conclusion The included MS patients have typical clinical manifestations, and their TCM constitution types are commonly seen as phlegm-damp constitution,damp-heat constitution,and Qi stagnation constitution. The results of constitution discrimination will supply evidence for the individualized therapy for the treatment and rehabilitation of MS patients.

Content Determination of 7 Constituents in Wikstroemia indica and Its Principal Component and Cluster Analysis / 中国药房

OBJECTIVE:To establish a method for simultaneous determination of 7 constituents in Wikstroemia indica,and to conduct principal component analysis and cluster analysis. METHODS:HPLC method was adopted. The determination was per-formed on Diamonsil Platisil ODS column with mobile phase consisted of acetonitrile-0.15% triethylamine solution(pH adjusted to 6.0 with phosphoric acid,gradient elution)at the flow rate of 1.0 mL/min. The detection wavelength was set at 280 nm,and col-umn temperature was 40 ℃. The sample size was 10 μL. The principal component analysis and cluster analysis were conducted for the results of content determination by SPSS 22.0 statistical software. RESULTS:The linear ranges were 2.688-53.76μg/mL for nar-ingin(r=0.9998),5.052-101.00 μg/mL for myricetin(r=0.9999),2.052-41.04 μg/mL for arctiin(r=0.9999),2.108-42.16 μg/mL(r=0.9999),5.112-102.20 μg/mL(r=0.9999),0.820-16.42 μg/mL(r=0.9999),2.070-41.40 μg/mL(r=0.9999),respec-tively. The limits of quantitation were no higher than 1.0720 μg/mL,the limits of detection were no higher than 0.3318 μg/mL. RSDs of precision,stability and reproducibility tests were all lower than 2.0%. The recoveries were 97.8%-102.5%(RSD=1.8%, n=6),97.2%-102.0%(RSD=2.0%,n=6),95.2%-100.1%(RSD=1.7%,n=6),95.2%-99.3%(RSD=1.6%,n=6), 97.0%-100.8%(RSD=1.3%,n=6),95.5%-98.6%(RSD=1.1%,n=6),95.0%-99.3%(RSD=1.8%,n=6),respectively. Three main components were belong to the samples of 10 batches of medicinal materials. The samples of medicinal materials from 10 pro-ducing area could be divided into 2 categories. The quality of W. indica from Qingyuan Guangdong and Guiyang Guizhou were bet-ter than others. CONCLUSIONS:The method is simple,precise,stable and reproducible,and it can be used for simultaneous de-termination of 7 constituents in medicinal material. The quality of W. indica from different regions are quite different.