Objective To explore the role of neogambogic acid in the characteristics of colorectal cancer stem cells (CRC-CSCs) through the Wnt/β-catenin signaling pathway. Methods The colorectal cells SW480 and HCT166 were divided into control group and neogambogic acid groups (1.5, 3, 6, and 12 μmol/L). The viability of CRC-CSCs was determined by MTT method, and spheroid and clone formation assays were used to assess the capacity of spheroid formation and self-renewal ability of the cells. The effects of neogambogic acid on the apoptosis and cell cycle of CRC-CSCs were evaluated by flow cytometry assays. Real-time quantitative PCR was used to detect the mRNA expression levels of relative markers (CD133, CD44, ALDH1, Oct4, and Nanog) of CRC-CSCs, and the protein expression levels of the self-renewal marker (PCNA), apoptosis markers (cleaved caspase-3 and cleaved caspase-9), and Wnt/β-catenin signaling pathway markers (p-GSK3β, GSK3β, β-catenin, and Wnt) were analyzed using Western blot. Results Compared with the control group, after neogambogic acid treatment, the viability of SW480 and HCT116 cells decreased (P<0.05), the spheroid forming ability and the clone numbers of CRC-CSCs decreased (P<0.001, P<0.01) but the cell apoptosis rate increased (P<0.01), and cell cycle was arrested in G₀/G₁ phase. Moreover, neogambogic acid downregulated the mRNA and protein expression of relative markers of CRC-CSCs (CD133, CD44, ALDH1, Oct4, and Nanog), PCNA, p-GSK3β, β-catenin, and Wnt (P<0.05) and upregulated the expression of cleaved caspase-3, cleaved caspase-9, and GSK3β (P<0.01). Conclusion Neogambogic can inhibit the stem cell properties of colorectal cells via inhibition of Wnt/β-catenin signaling pathway. As a result, neogambogic acid may be an attractive agent against colorectal cancer.

Objective:To observe the effect of warming needle therapy combined with Tuina(Chinese therapeutic massage)on the clinical symptoms and anxiety state of patients with cervical vertigo accompanied by anxiety. Methods:Seventy patients with cervical vertigo accompanied by anxiety state were divided into an observation group and a control group using the random number table method,with 35 patients in each group.In addition to disease education,the observation group was treated with warming needle therapy and Tuina 3 times a week,and the control group was treated with betahistine mesylate tablets orally 3 times a day.Both groups were treated for 4 weeks.The changes in the scores of the evaluation scale for cervical vertigo(ESCV),self-rating anxiety scale(SAS),and Hamilton anxiety rating scale(HAMA)were compared between the two groups after treatment. Results:One case in the observation group dropped out due to failure to cooperate with the treatment during the study.In the treatment group,the total effective rate and the cured plus markedly effective rate were 94.1%and 50.0%,respectively,and 88.6%and 8.6%in the control group,respectively;the differences between the two groups were statistically significant(P<0.05).After treatment,the ESCV,SAS,and HAMA scores of both groups decreased significantly compared with those before treatment(P<0.01),and the scores of the observation group were lower than those of the control group(P<0.01 or P<0.05). Conclusion:Warming needle therapy combined with Tuina can significantly improve the clinical symptoms of patients with cervical vertigo accompanied by anxiety state.

Objective:To summarize the publications in the field of neurofibroma and malignant peripheral nerve sheath tumor in the past 25 years, to illuminate the cooperation among countries, academic institutions and researchers, and to compare the research topics between Chinese and English databases, and to predict the research trend.Methods:Web of Science and CNKI databases were searched, and literature about neurofibroma and malignant peripheral nerve sheath tumor(MPNST) resection from January 1st, 1997 to September 2nd, 2021 were included. Excel 2017 was used for statistical analysis. VOSviewer 1.6.16 software was used for clustering and visualization of countries, institutions, authors, and keywords.Results:A total of 9 620 English articles and 957 Chinese articles were included. Over the past 25 years, Chinese and English publications have been increasing quickly. In terms of countries and institutions, most of the countries with a large number of documents published in English literature (top 10) are developed countries and the institutional cooperation between them is relatively close. In terms of keywords clustering, the result of English literature are more comprehensive, including the pathogenesis, clinical manifestations, diagnosis and treatment of complications. The clustering result suggest that the potential research hotspots in the future may be the impact of tumor microenvironment on the development of neurofibroma and MPNST, molecular typing of neurofibroma, drug resistance mechanism of neurofibroma and so on. The clustering of Chinese literature can be summarized as the auxiliary diagnosis of neurofibroma and the treatment of complications.Conclusions:In the future, researchers may pay more attention to the role of tumor microenvironment in the development of neurofibroma, driving factors of neurofibroma and MPNST, molecular typing and drug resistance mechanism of neurofibroma.

Objective:To summarize the publications in the field of neurofibroma and malignant peripheral nerve sheath tumor in the past 25 years, to illuminate the cooperation among countries, academic institutions and researchers, and to compare the research topics between Chinese and English databases, and to predict the research trend.Methods:Web of Science and CNKI databases were searched, and literature about neurofibroma and malignant peripheral nerve sheath tumor(MPNST) resection from January 1st, 1997 to September 2nd, 2021 were included. Excel 2017 was used for statistical analysis. VOSviewer 1.6.16 software was used for clustering and visualization of countries, institutions, authors, and keywords.Results:A total of 9 620 English articles and 957 Chinese articles were included. Over the past 25 years, Chinese and English publications have been increasing quickly. In terms of countries and institutions, most of the countries with a large number of documents published in English literature (top 10) are developed countries and the institutional cooperation between them is relatively close. In terms of keywords clustering, the result of English literature are more comprehensive, including the pathogenesis, clinical manifestations, diagnosis and treatment of complications. The clustering result suggest that the potential research hotspots in the future may be the impact of tumor microenvironment on the development of neurofibroma and MPNST, molecular typing of neurofibroma, drug resistance mechanism of neurofibroma and so on. The clustering of Chinese literature can be summarized as the auxiliary diagnosis of neurofibroma and the treatment of complications.Conclusions:In the future, researchers may pay more attention to the role of tumor microenvironment in the development of neurofibroma, driving factors of neurofibroma and MPNST, molecular typing and drug resistance mechanism of neurofibroma.

Objective:To explore the correlation of the expression of lymphocyte immunoglobulin-mucin domain 3 (Tim-3) on T lymphocytes and natural killer (NK) cells with hepatic inflammation and hepatic fibrosis in patients with chronic hepatitis B virus (HBV) infection.Methods:A total of 320 patients of chronic HBV infection who visited the Infectious Diseases Department in the Third Affiliated Hospital of Sun Yat-sen University from June 2016 to June 2018 were enrolled. The patients were divided into four groups: immune tolerant group (IT, n=31), immune active group (IA, n=184), inactive carriers group (IC, n=48), and gray zone group (GZ, n=57). And 17 healthy controls (HC group) were included at the same time. Peripheral blood mononuclear cells were separated and the frequency and mean fluorescence intensity (MFI) of Tim-3 on T cells (CD3 + , CD4 + and CD8 + T cells) and NK cells (NK, NK-bright and NK-dim cells) were detected by flow cytometry. The clinical data of patients were collected and aspartate aminotransferase-to-platelet ratio index (APRI) score was calculated. Kruskal-Wallis H test was used for comparing the data of non-normal distribution among groups, and Mann Whitney U test was used for the comparison between two groups. Enumeration data were expressed as cases (percentage) and compared by the Chi-square test. Spearman rank correlation was used for correlation analysis. Receiver operating characteristic curve (ROC) was used to analyze the predictive value of Tim-3 expression on T cells and NK cells in evaluating liver fibrosis in patients with chronic HBV infection. P<0.05 was considered statistically significant. Results:Significant differences were found in the age, aspartate aminotransferase (AST), alanine aminotransferase(ALT), albumin (Alb), total bilirubin (TBil) and liver stiffness measurement (LSM) among IT, IA, IC, GZ and HC groups ( H=12.40, 169.70, 210.70, 25.17, 24.21 and 86.5, all P<0.05). And the differences in APRI score, proportion of HBeAg-positive patients, HBsAg and HBV-DNA among the IT group, IA group, IC group, GZ group were also significant ( H=89.45, 118.00 and 14.81, χ2=148.20, all P<0.05). The frequency and MFI of Tim-3 on CD3 + , CD4 + and CD8 + T cells, NK cells, NK-bright and NK-dim cells among the IT group, IA group, IC group, GZ group and the HC group were significantly different( H=13.57, 51.55, 8.58, 44.25, 20.32, 47.96 and 12.45, 33.69, 4.96, 32.47, 10.63, 30.46, all P<0.05). Both of the frequency and MFI of Tim-3 on CD3 + , CD4 + and CD8 + T cells were positively correlated with ALT and AST levels in patients with chronic HBV infection ( r=0.2134, 0.4733, 0.2090, 0.4333, 0.1771, 0.4417, 0.1780, 0.3956, 0.2618, 0.4671, 0.2614 and 0.4326, all P<0.05). While the frequency and MFI of Tim-3 on CD8 + T cells and MFI on CD3 + and CD4 + T cells were also positively correlated with TBil levels ( r=0.1342, 0.2635, 0.2739 and 0.2526, all P< 0.05). The frequency and MFI of Tim-3 on NK and NK-dim cells were negatively correlated with the levels of ALT, AST and TBil ( r=-0.2671, -0.4093, -0.2451, -0.4099, -0.1807, -0.1823, -0.2733, -0.4224, -0.2576, -0.4206, -0.1798 and -0.1946, all P<0.05). The MFI of Tim-3 on NK-bright cells was also negatively correlated with ALT, AST and TBil ( r=-0.3775, -0.3562 and -0.1633, all P<0.05). Both of the frequency and MFI of Tim-3 on CD3 + , CD4 + and CD8 + T cells were positively correlated with liver fibrosis( r=0.1789, 0.3896, 0.1518, 0.3521, 0.2117 and 0.3579, all P<0.05). Both of the frequency and MFI of Tim-3 on CD4 + and CD8 + T cells and the MFI of Tim-3 on CD3 + T cells were positively correlated with APRI score ( r=0.1487, 0.2604, 0.2296, 0.4858 and 0.2853, all P<0.05). The expression frequency and MFI of Tim-3 on NK and NK-dim cells and MFI of Tim-3 on NK-bright cells were negatively correlated with LSM ( r=-0.2686, -0.3975, -0.2852, -0.3991 and -0.3531, all P<0.05). The expression frequency and MFI of Tim-3 on NK and NK-dim cells and MFI of Tim-3 on NK-bright were negatively correlated with APRI score ( r=-0.3589, -0.4158, -0.3591, -0.4108 and -0.3966, all P<0.05). The ratio of Tim-3 expression on CD3 + T cells to that on NK cells was shown to be able to predict liver fibrosis in chronic HBV infected patients and the area under the ROC curve was 0.783 (95% CI: 0.723~0.843, P< 0.05), and when the cut-off value was 0.612, the sensitivity was 61.9%, and the specificity was 99.3%. Conclusion:The relationship of Tim-3 expression on T cells with liver inflammation and fibrosis is opposite to that on NK cells in patients with chronic HBV infection, indicating that the ratio of Tim-3 expression on T cells to that on NK cells may be valuable in evaluating liver fibrosis in patients.

Objective: To analyze the changes and significance of serum soluble programmed death-1 (sPD-1) in patients with chronic hepatitis C (CHC) and study its role in the progression of CHC. Methods: Serum levels of sPD-1 in CHC patients and healthy controls (HC) were measured using ELISA and compared. Correlations of serum sPD-1 with peripheral hepatitis C virus (HCV) RNA, alanine aminotransferase (ALT), aspartate aminotransferase (AST) and liver fibrosis (indicated by APRI) were analyzed. Results: The serum sPD-1 level in the CHC group was significantly higher than that in the HC group (P<0.05), and positively correlated with peripheral HCV RNA, ALT and AST (P<0.05). In addition, the serum sPD-1 levels in patients with APRI greater than 1.2 (indicating severe liver fibrosis) were higher than those in patients without or with mild liver fibrosis (P<0.05). Conclusions sPD-1 might be involve in the progression of CHC. Measuring serum sPD-1 in CHC patients would assist the prediction of disease progression and help to make the correct diagnosis and appropriate clinical therapy decision.

Objective To compare the clinical effect of two kinds of internal fixation of locking plate and anatomical plate in the treatment of Pilon fracture.Methods 64 patients with Pilon fracture undergoing elective surgery were randomly divided into study group and control group by digital table method,32 cases in each group.The study group received locking plate internal fixation for the treatment,the control group was treated with anatomical plate internal fixation.The clinical efficacy and safety of two kinds of internal fixation methods were compared.Results After treatment,the excellent rate of ankle joint function in the study group(96.88%)was significantly higher than that in the control group (81.25%),and the difference had statistical ssignificance (χ2 =4.01,P <0.05 ).The operative time,hospitalization time and fracture healing time[(57.32 ±10.98)min,(13.72 ±591 )d,(15.72 ± 2.82)weeks]in the study group were shorter than those in the control group[(68.11 ±11.85 )min,(16.62 ± 5.21)d,(17.81 ±3.42)weeks],and the differences had statistical significance(t =3.98,3.11,321,all P <0.05). The amount of bleeding (153.32 ±21.36)mL in the study group was less than that in the control group [(188.65 ± 26.24)mL](t =5.46,P <0.05).The incidence rate of delayed fracture healing of the study group (3.13%)was significantly lower than that in the control group(28.13%),the difference had statistical significance(χ2 =7.59,P <0.05).There were no significant differences in the incidence of other complications between the two groups(χ2 =1.95,102,102,0.35,all P >0.05).Conclusion Locking plate fixation in the treatment of Pilon fractures could significantly promote the recovery of joint function,and has the advantages of short operation time,fast fracture healing,less complications and other advantages,it is better than anatomical plate internal fixation.

Stroke is responsible for increasingly high rates of mortality and disability worldwide. Approximately two million people suffer from stroke for the first time in China each year. The high incidence (50%) of post-stroke disability brings a heavy burden to patients and their caregivers. Acupuncture has been widely used in the communities for post-stroke rehabilitation in China. The objective of this trial is to apply our acupuncture research achievement to treatment and evaluation of post-stroke hemiplegic patients in community.

A cidithiobacillus ferrooxidans is a chemolithotrophic microorganism capable of using ferrous ions and sulphides as energy sources. This microorganism has an important role in the bioleaching of minerals. During this process, the bacteria are normally subjected to several stressing conditions, such as temperature changes, lack of nutrients or pH changes, which may affect the efficiency of the bacterial action. SELDI is a recent technology that allows for high-throughput proteomics studies. The Protein Chip SELDI technology was used to generate comparative protein profiles of A c idithiobacillus ferrooxidans grown under phosphate starvation or normal condition additionally adding Fe2+ as energy resource. There were 13 significantly differential expressed protein's peaks found by using SELDI Protein Chip technologies, which made a solid foundation for further isolation these low molecular proteins by adopting technologies such as HPLC etc.

Aim To establish a HPLC method for determination of the content of Huperzine A in Huperzia serrata and compare it in different parts of the plants. Method DiamonsilTm C_ 18 column(200 mm?4.6 mm,5 ?m)was used; the mobile phase was methanol ∶0.08 mol?L-1ammonium acetate buffer(40 ∶60,pH=6.0),the detection wavelength was 308 nm, with flow rate at 1.0 ml?min-1;the column temperature was room temperature.Results There is a good linear relationship within the range of 5～100 mg?L-1(r=0.9997).The average recovery was 99.10%,with RSD 2.44%(n=5).Regarding the content of Huperzine A in different parts of Huperzia serrata, it was found that the quantity of Huperzine A in leaf was higher than that in stem and it was the lowest in root. Conclusion The method is simple, accurate and reliable etition. Huperzine A can be accumulated in the aboveground part of the plant.