Objective To investigate the effects of Echinococcus multilocularis infection on levels of memory T (Tm) cells and their subsets in lymph nodes of mice at different stages of infection, so as to provide new insights into immunotherapy for alveolarechinococcosis. MethodsTwenty-four C57BL/6J mice aged 6 to 9 weeks were randomly divided into the infection group and the control group, of 12 mice in each group. Mice in the infection group were administered with 3 000 E. multilocularis protoscoleces via portal venous injection, while animals in the control group were administered with an equal volume of physiological saline. Three mice from each group were sacrificed 4, 12 weeks and 24 weeks post-infection, and lymph nodes were sampled and stained with hematoxylin and eosin (HE) to investigate the histopathological changes of mouse lymph nodes in the infection group. The expression and localization of T lymphocyte surface markers CD3, CD4, and CD8 were observed in mouse lymph nodes using immunohistochemical staining. In addition, lymphocyte suspensions were prepared from mouse lymph nodes in both groups at different time points post-infection, and the levels of Tm cell subsets and their secreted cytokines were detected using flow cytometry. Results HE staining showed diffuse structural alterations in the subcapsular cortical and paracortical regions of mouse lymph nodes in the infection group 4 weeks post-infection with E. multilocularis. Immunohistochemical staining detected CD3, CD4 and CD8 expression in mouse lymph nodes in both groups. Flow cytometry revealed higher proportions of CD4⁺ Tm cells [(55.3 ± 4.8)% vs. (38.8 ± 6.1)%; t = -4.259, P < 0.05] and CD4⁺ tissue-resident Tm (Trm) cells [(57.7 ± 3.7)% vs. (34.1 ± 11.2)%; t = -3.990, P < 0.05] in mouse lymph nodes in the infection group than in the control group 4 weeks post-infection, and higher proportions of CD4⁺ Tm cells [(34.6 ± 3.2)% vs. (23.3 ± 7.5)%; t = -2.764, P < 0.05] and CD4⁺ Trm cells [(44.0 ± 1.9)% vs. (31.2 ± 1.5)%; t = -4.039, P < 0.05] in mouse lymph nodes in the infection group than in the control group 24 weeks post-infection. The proportions of CD8⁺ Tm cells were higher in the infection group than in the control group 4 weeks [(56.8 ± 2.7)% vs. (43.9 ± 5.2)%; t = -4.416, P < 0.01] and 12 weeks post-infection [(25.4 ± 2.7)% vs. (12.0 ± 2.6)%; t = -2.552, P < 0.05], while the proportions of tumor necrosis factor (TNF)-α⁺ CD4⁺ T cells [(15.7 ± 5.0)% vs. (49.4 ± 6.4)%; t = 7.150, P < 0.01], TNF-α⁺CD8⁺ T cells [(20.7 ± 5.5)% vs. (57.5 ± 8.4)%; t = -6.694, P < 0.01], and TNF-α⁺ CD8⁺ Tm cells [7.0% (1.0%) vs. 31.0% (11.0%); Z = -2.236, P < 0.05] were lower in the infection group than in the control group 24 weeks post-infection. Conclusions Tm cells levels are consistently increased in lymph nodes of mice at different stages of E. multilocularis infection, with Trm cells as the predominantly elevated subset. The impaired capacity of CD8⁺ Tm cells to secrete the effector molecule TNF-α in mouse lymph nodes at the late-stage infection may facilitate chronic parasitism of E. multilocularis.

Objective To investigate the effect of LAG-3 deficiency (LAG3^-/-) on natural killer (NK) cell function and hepatic fibrosis in mice infected with Echinococcus multilocularis. Methods C57BL/6 mice, each weighing (20 ± 2) g, were divided into the LAG3^-/- and wild type (WT) groups, and each mouse in both groups was inoculated with 3 000 E. multilocularis protoscoleces via the hepatic portal vein. Mouse liver and spleen specimens were collected 12 weeks post-infection, sectioned and stained with sirius red, and the hepatic lesions and fibrosis were observed. Mouse hepatic and splenic lymphocytes were isolated, and flow cytometry was performed to detect the proportions of hepatic and splenic NK cells, the expression of CD44, CD25 and CD69 molecules on NK cell surface, and the secretion of interferon γ (IFN-γ), tumor necrosis factor α (TNF-α), interleukin (IL)-4, IL-10 and IL-17A. Results Sirius red staining showed widening of inflammatory cell bands and hyperplasia of fibrotic connective tissues around mouse hepatic lesions, as well as increased deposition of collagen fibers in the LAG3-/-group relative to the WT group. Flow cytometry revealed lower proportions of mouse hepatic (6.29% ± 1.06% vs. 11.91% ± 1.85%, P < 0.000 1) and splenic NK cells (4.44% ± 1.22% vs. 5.85% ± 1.10%, P > 0.05) in the LAG3^-/- group than in the WT group, and the mean fluorescence intensity of CD44 was higher on the surface of mouse hepatic NK cells in the LAG3^-/- group than in the WT group (t = −3.234, P < 0.01), while no significant differences were found in the mean fluorescence intensity of CD25 or CD69 on the surface of mouse hepaticNK cells between the LAG3^-/- and WT groups (both P values > 0.05). There were significant differences between the LAG3^-/- and WT groups in terms of the percentages of IFN-γ (t = −0.723, P > 0.05), TNF-α (t = −0.659, P > 0.05), IL-4 (t = −0.263, P > 0.05), IL-10 (t = −0.455, P > 0.05) or IL-17A secreted by mouse hepatic NK cells (t = 0.091, P > 0.05), and the percentage of IFN-γ secreted by mouse splenic NK cells was higher in the LAG3^-/- group than in the WT group (58.40% ± 1.64% vs. 50.40% ± 4.13%; t = −4.042, P < 0.01); however, there were no significant differences between the two groups in terms of the proportions of TNF-α (t = −1.902, P > 0.05), IL-4 (t = −1.333, P > 0.05), IL-10 (t = −1.356, P > 0.05) or IL-17A secreted by mouse splenic NK cells (t = 0.529, P > 0.05). Conclusions During the course of E. multilocularis infections, LAG3^-/- promotes high-level secretion of IFN-γ by splenic NK cells, which may participate in the reversal the immune function of NK cells, resulting in aggravation of hepatic fibrosis.

ObjectiveUltra performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry（UPLC-Q-Exactive Orbitrap MS） was used to analyze the differences in chemical components between raw products and wine-processed products of Polygonatum cyrtonema rhizome. MethodUPLC-Q-Exactive Orbitrap MS was used to analyze the chemical compositions of P. cyrtonema rhizome before and after processing， and the effective response ions were extracted after raw data processing， and the differential compounds before and after processing were screened combined with multivariate statistical analysis and according to the conditions of variable importance in the projection（VIP） value>1， P<0.05， fold change（FC）>2 or FC<0.5， based on the retention time， quasi-molecular ions， fragment ions and other information， the components were identified in combination with the control products and the literature， and the significant difference compounds were identified by clustering thermal analysis and relative quantitative analyzed， in order to clarify the change rule of the main components in P. cyrtonema rhizome before and after processing. ResultA total of 72 differential constituents between raw products and wine-processed products were identified， including 15 alkaloids， 12 organic acids， 12 amino acids， 6 flavonoids， 4 saccharides and 23 others. There were a total of 18 significantly different components， among which 13 compounds， including L-malic acid， lactic acid and 9，12，13-trihydroxy-10-octadecenoic acid， showed an increasing trend in content after wine processing， 5 compounds such as trans-3-indoleacrylic acid， L-arginine， D-tryptophan， showed a decreasing trend after processing. ConclusionThe chemical components of P. cyrtonema rhizome are significantly different before and after processing， mainly organic acids， saccharides， amino acids， flavonoids and alkaloids， which can lay the foundation for the in-depth study of the processing mechanism of Polygonati Rhizoma.

Objective:To investigate the immune cell infiltration width on lesion microenvironment (LME) based on different hepatic alveolar echinococcosis lesion features to underlie referential sampling range for experimental and control tissues aiming at avoiding false negativity in basic researches.Methods:Using prospective research methods, from January 2017 to December 2019, patients with hepatic alveolar echinococcosis who were diagnosed and treated surgically at the First Affiliated Hospital of Xinjiang Medical University and met the multi-site sampling method (MSS method) were investigated. A total of 26 cases were included, aged 34 (15, 65) years old, with a gender ratio of 12/14. Lesions were classified into six groups based on heterogenic scales of calcification and liquefaction: A. non-calcified and non-liquefied; B. obvious calcified and non-liquefied; C. partial calcified and partial liquefied; D. obvious calcified and partial liquefied; E. partial calcified and subtotal liquefied; F. obvious calcified and subtotal liquefied. Liver specimens were acquired with 5 mm interval off the lesion shore in LME area using MSS method. Performed immunohistochemical staining of CD3, CD19, CD68, and Masson staining of fibrous tissue, and after pathological evaluation, the layer with a sharp decrease in immune cell infiltration abundance was determined as the maximum infiltration range (width, W value). The experimental group conservatively estimated the maximum sampling range for the integer value of Q1 of W value. The control group conservatively estimated the minimum sampling range for the integer value after Q3 rounding plus 5 mm of W value. Results were analyzed using Kruskal-Wallis H and Mann-Whitney U tests, referential ranges were concluded. Results:Median W values (interquartile) for each group were 20.0 (12.5, 22.5), 15.0 (10.0, 15.0), 10.0 (10.0, 15.0), 5.0 (5.0, 10.0), 12.5 (6.3, 15.0), and 5.0 (5.0, 10.0) mm, among which A > D, A > F, C > D, and C > F ( P≤0.05); from the perspective of calcification, A > C + E, A > B + D + F ( P < 0.05), while A + B > C + D, A + B > E + F ( P < 0.05) from the perspective of liquefaction. In these groups, the experimental group conservatively estimated the maximum distance for sampling to be 12.0, 10.0, 10.0, 5.0, 6.0, and 5.0 mm, while the control group conservatively estimated the minimum distance for sampling to be 28.0, 20.0, 20.0, 15.0, 20.0, and 15.0 mm. Conclusions:Less calcification and liquefaction implicates wider immune cell infiltration range in those lesions. Tissue sampling should be individualized based on different lesion features in basic research to avoid false negativity.

Objective:To investigate characteristics of the 18F-flurodeoxyglucose ( 18F-FDG) uptake intensity and ranges in distinct hepatic alveolar echinococcosis lesions. Methods:The clinical data of 39 patients with position emission tomography during Jan 2017 to Dec 2019 in the First Affiliated Hospital of Xinjiang Medical University were enrolled. Among them, there were 17 males and 22 females, aging from 15 to 65 years (median 34 years). Lesions were classified into six groups based on heterogenic scales of calcification and liquefaction: A. non-calcified and non-liquefied ( n=7); B. obvious calcified and non-liquefied ( n=7); C. partial calcified and partial liquefied( n=10); D. obvious calcified and partial liquefied ( n=5); E. partial calcified and subtotal liquefied ( n=5); F. obvious calcified and subtotal liquefied ( n=5). Tumor to background ratio (TBR) and width (W) of lesion infiltrative boundary were measured and calculated. Statistical comparison using Mann-Whitney U test as well as correlation analysis was performed. Results:TBR values [ M( Q1, Q3)] for each group were 4.40(3.66, 7.03), 2.55(1.69, 3.60), 3.73(3.37, 5.21), 2.90(2.75, 3.60), 3.80(3.49, 6.36), 2.49(2.21, 3.97), among which A>B, A>D, A>F, C>B, E>B ( U=3.0, 4.0, 4.5, 11.0, 5.0, all P<0.05); From the perspective of the calcification in each group, it was found that the lighter the calcification was, the greater the TBR value was. W values [ M( Q1, Q3)] for each group were [12.5(10.0, 19.5), 11.2(10.5, 12.5), 12.2(10.9, 13.2), 7.8(7.3, 9.3), 10.0(7.3, 13.4), 7.3(6.8, 7.6)] mm, among which A>D, A>F, B>D, B>F, C>D, C>F (all U=0, all P<0.05); According to the degree of calcification and liquefaction of lesions in each group, the lighter the calcification was, the greater the W value was; The heavier the liquefaction was, the smaller the W value was. A mild strength linear correlation has been observed between the TBR value and W value ( r=0.4136, P<0.05). Conclusions:Less calcification and liquefaction implicated higher 18F-FDG uptake intensity and wider range. Radical resection margins and tissue sampling should be individualized based on different lesion features in surgical treatment.

OBJECTIVE：To establish the m ethod for simultaneous determination of 8 kinds of ginsenosides in Panax quinquefolium broken pieces. METHODS ：HPLC-DAD method was used to determine the contents of ginsenoside Rg 1，Re，Rb1， Rc，Ro，Rb2，Rb3，Rd in P. quinquefolium broken pieces. The determination was performed on Agilent 5 TC-C18 column with mobile phase consisted of acetonitrile- 0.2％ phosphoric acid water solution （gradient elution ）at the flow rate of 1.0 mL/min. The column temperature was set at 30 ℃. The detection wavelength was set at 203 nm，and sample size was 10 μL. Ginsenoside Re and ginsenoside Rb 2 were used as control ，liner calibration with two-reference substances correction was used to predict the retention time of other 6 components，and was compared with the relative retention time method. Using ginsenoside Re as control ， above components were quantified by the relative correction factor method ，and the results were compared with the external standard method. RESULTS ：The contents of ginsenoside Rg 1，Re，Rb1，Rc，Ro，Rb2，Rb3，Rd were 10.59-12.78，2.160-2.768， 27.492-38.880，3.154-4.018，3.368-4.080，0.343-0.755，0.961-1.415，5.857-6.923 mg/g. The accuracy of two-reference substances linear correction method to predict the retention time of components was higher ，and the absolute deviation of the predicted retention time was lower than that of the relative retention time method. There was no significant difference between the relative correction factor method and the external standard method ，and relative error was ＜3％ . CONCLUSIONS ：Established two-reference substances for determination of multiple components can be used for qualitative and quantitative analysis of 8 kinds of ginsensides in P. quinquefolium broken pieces simultaneously and accurately.

Human alveolar echinococcosis is a chronic infectious disease caused by Echinococcus multilocularis infection. It predominantly injuries the liver and grows like the malignant tumor. The therapeutic options and prognosis depend on types of human alveolar echinococcosis, clinical stages, biological activity, vascular invasion, pathological characteristics, and patient's immune status. However, despite of multiple classification methods, there are still lacking of comprehensive typing schemes. which leads to inappropriate diagnosis and therapy. This research systematically reviewed the recent studies on human alveolar echinococcosis at home and abroad and analyzed the classifications based on ultrasound, computer tomography, magnetic resonance imaging, positron emission computed tomography, serology and pathology, and some novel technologies and summarized the individual advantage and disadvantage for each classification Relationships and their advantages plus disadvantages have been assessed comprehensively. Meanwhile, the possible reference factors or theoretical basis for optimized future classification are proposed, in order to establish a unified classification system to provide guidance for clinical diagnosis and treatment.

OBJECTIVE：To establish the content determination me thod of 3 mono/disaccharides in 3 kinds of medicinal Dendrobii Caulis. METHODS ：HPLC-CAD method was established. The determination was performed on Shodex Asahipak NH2P-50 4E column with mobile phase consisted of acetonitrile-water （75 ∶ 25，V/V）at the flow rate of 1.0 mL/min. The column temperature was set at 30 ℃，and sample size was 10 µL. CAD detection condition included that data acquisition frequency was 5 Hz，filter constant was 5 s，atomization temperature was 35 ℃ ，gas source was nitrogen with pressure of 4.012 × 105 Pa. RESULTS：The linear range of fructose ，D-anhydrous glucose and sucrose were 0.156 2-1.873 8 mg/mL（r＝0.999 5），0.012 7- 0.152 4 mg/mL（r＝0.999 7），0.277 6-3.331 2 mg/mL（r＝0.999 8），respectively. The limits of quantification were 0.002 61，0.004 24 and 0.005 12 mg/mL，and the limits of detection were 0.000 78，0.001 27 and 0.001 54 mg/mL，respectively. RSDs of precision ， stability，reproducibility and durability tests were all lower than 3％. The recoveries were 95.98％-98.15％（RSD＝0.83％，n＝6）， 95.64％-98.62％（RSD＝1.10％，n＝6）and 97.53％-98.94％（RSD＝0.53％，n＝6）. The contents of them were 0.28％-1.12％， 0.02％-0.13％，0.76％-2.67％，respectively. The total content was 1.38％～3.10％. The order of saccharide content in 3 kinds of Dendrobii Caulis was sucrose ＞fructose＞D-anhydrous glucose ；the order of sucrose content and total content were Dendrobium huoshanense＞D. moniliforme ＞D. officinale ；the order of D-anhydrous glucose content was D. huoshanense ＞D. officinale ＞D. moniliforme； the order of fructose content was D. moniliforme ＞D. officinale ＞D. huoshanense . CONCLUSIONS ：Established method is sensitive ，reproducible and simple in operation ，and can be used for content determination of 3 saccharides in 3 kinds of medicinal Dendrobii Caulis. There are differences in the contents of saccharide s among 3 kinds of Dendrobii Caulis.

Objective To compare the clinical efficacy and postoperative liver function in patients with primary hepatic carcinoma treated by transcatheter arterial chemoembolization(TACE) or TACE combined with portal vein chemoembolization.Methods 48 patients with primary hepatic carcinoma, randomly divided into 2 groups (hepatic artery group in 25 cases and dual interventional group in 23 cases),underwent interventional treatment.The hepatic artery group underwent conventional hepatic artery interventional therapy, while the dual interventional group underwent hepatic artery and portal vein interventional treatment.The postoperative clinical efficiency, liver volume and liver function between the two groups'' patients were compared.Results To the endpoint of observation,the clinical efficacy and tumor reduction degree of dual interventional group were better than that of hepatic artery group.Compared with hepatic artery group, the postoperative ALT, AST and TBIL of dual interventional group were higher on the first and third days.On the seventh and fourteenth days, the statistical difference was not significant.The volume of non-embolization part in dual interventional group was larger than that in preoperative volume to different degrees.The most obvious change of liver volume happened in the 4th weeks after treatment.There was no treatment-related death or severe adverse reaction in two groups.Conclusion The treatment of TACE combined with portal vein chemoembolization is a safe and effective method, which may effectively inhibit the growth and reduce the volume of tumor, and result in compensatory hypertrophy of non-embolization part.

Objective:To detect the expression and clinical significance of β-tubulin Ⅲ in cancer tissue of the patients with colon adenocarcinoma,and to explore its clinical significance.Methods:A total of 111 colon adenocarcinoma tissue samples were obtained.According to the location of β-tubulin Ⅲ positive cells, all patients were divided into front group (n=72) and non-front group (n=39).The positive expression rate of β-tubulin Ⅲ in the patients with colon adenocarcinoma was detected with immunohistochemistry.The correlations among the expression of β-tubulin Ⅲ and gender,age,tumor differentiation, clinical stage, lymph node metastasis, recurrence and death were analyzed.Results: The expression levels of β-tubulin Ⅲ had no significant differences between the patients with different gerder,age,lymph node metastasis,clinical stages,death and recurrence.The positive expression rates of β-tubulin in cancer tissue of the patients had significant difference between front and non-front groups (χ2=8.76, P=0.01).Lowly-to-moderately differentiated tissue was more common in front group, and highly-differentiated tissue was more common in non-front group(χ2=6.88, P=0.03).There were significant differences in the expression levels of β-tubulin Ⅲ between cancer tissues with different differentiation degrees (χ2=5.74, P=0.04).In non-front group, lymph node metastasis was closely correlated with the expression of β-tubulin Ⅲ (χ2=6.02,P=0.05).The results of immunohistochemical staining showed that the β-tubulin Ⅲ positive-expressing cells were colored brown-yellow.The number of cells with positive β-tubulin Ⅲ expression was significantly increased in highly differentiated tissue compared with low-differentiated tissue.Conclusion:The expression of β-tubulin Ⅲ is closely related to tumor differentiation in colon adenocarcinoma tissue.The highly differentiated colon adenocarcinoma tissue is more common in non-front group in which the expression of β-tubulin Ⅲ is related to lymph node metastasis.