ObjectiveTo explore effect of Xianlian Jiedu prescription on the recurrence of colorectal cancer （CRC） and investigate the related mechanisms. MethodsA postoperative recurrence model was established in 25 Balb/c mice by injecting CT26 cells subcutaneously into the armpit， followed by surgical removal of 99% of the subcutaneous tumor. The mice were randomly divided into model group， low-dose Xianlian Jiedu prescription （XLJDP-L） group （6.45 g·kg^-1·d^-1）， medium-dose Xianlian Jiedu prescription （XLJDP-M） group （12.9 g·kg^-1·d^-1）， high-dose Xianlian Jiedu prescription （XLJDP-H） group （25.8 g·kg^-1·d^-1）， and 5-fluorouracil （5-FU） group （1×10^-3 g·kg^-1·d^-1）. The mice were euthanized after 14 days of continuous intervention， and recurrent tumor tissue was harvested. Hematoxylin and eosin （HE） staining was used to observe pathological and morphological changes in the recurrent tumor tissue. Immunohistochemistry （IHC） was employed to assess the expression of proliferating cell nuclear antigen （Ki67）， vascular endothelial growth factor （VEGF）， and platelet-endothelial cell adhesion molecule （CD31） in recurrent tumor tissue. The Western blot was used to detect the protein expression levels of angiopoietin-2 （ANG-2）， VEGF， phosphorylated-protein kinase B （p-Akt）， protein kinase B （Akt）， phosphorylated-phosphatidylinositol 3-kinase （p-PI3K）， and phosphatidylinositol 3-kinase （PI3K） in recurrent tumor tissue. ResultsBefore treatment， there were no statistical differences in tumor volume， tumor weight， and body mass among the XLJDP-L， XLJDP-M， and XLJDP-H groups and the 5-FU group compared to the model group， indicating model stability. After treatment， compared with those in the model group， the tumor volume and tumor weight in the XLJDP-L， XLJDP-M， and XLJDP-H groups and the 5-FU group were significantly reduced （P<0.01）， showing dose dependency. Meanwhile， there were no significant differences in body weight among the XLJDP-L， XLJDP-M， and XLJDP-H groups and the 5-FU group compared to the model group. HE staining showed that compared with that in the model group， tumor tissue in the XLJDP-L， XLJDP-M， and XLJDP-H groups and the 5-FU group had loosely arranged cells， increased intercellular spaces， small and shriveled nuclei， light staining， fewer mitotic figures and atypical nuclei， and increased necrotic areas. IHC showed that compared with those of the model group， the positive rates of Ki67， VEGF， and CD31 in the recurrent tumor tissue of the XLJDP-L， XLJDP-M， and XLJDP-H groups and the 5-FU group were significantly reduced （P<0.01） in a dose-dependent manner. Western blot results showed that compared with those of the model group， the protein expression levels of ANG-2 and VEGF in the recurrent tumor tissue of the XLJDP-L， XLJDP-M， and XLJDP-H groups and the 5-FU group were significantly downregulated （P<0.05， P<0.01）， and the p-Akt/Akt and p-PI3K/PI3K ratios were significantly decreased in a dose-dependent manner （P<0.05， P<0.01）. ConclusionXianlian Jiedu prescription significantly inhibits the recurrence of CRC in mice after subcutaneous tumor surgery. The mechanism may involve regulating the PI3K/Akt pathway and downregulating key angiogenic proteins such as ANG-2， VEGF， and CD31.

Colorectal tumorigenesis generally progresses from adenoma to adenocarcinoma, accompanied by dynamic changes in the tumor microenvironment (TME). A randomized controlled trial has confirmed the efficacy and safety of Shen-Bai-Jie-Du decoction (SBJDD) in preventing colorectal tumorigenesis. However, the mechanism remains unclear. In this study, we employed single-cell RNA sequencing (scRNA-seq) to investigate the dynamic evolution of the TME and validated cell infiltration with multiplex immunohistochemistry and flow cytometry. Bulk RNA sequencing was utilized to assess the underlying mechanisms. Our results constructed the mutually verifiable single-cell transcriptomic atlases in Apc ^Min/+ mice and clinical patients. There was a marked accumulation of CCL22⁺ dendritic cells (DCs) and an enhanced immunosuppressive action, which SBJDD and berberine reversed. Combined treatment with cholesterol and lipopolysaccharide induced characteristic gene expression of CCL22⁺ DCs, which may represent "exhausted DCs". Intraperitoneal injection of these DCs after SBJDD treatment eliminated its therapeutic effects. TMEM131 derived CCL22⁺ DCs generation by TNF signaling pathway and may be a potential target of berberine in retarding colorectal tumorigenesis. These findings emphasize the role of exhausted DCs and the regulatory mechanisms of SBJDD and berberine in colorectal cancer (CRC), suggesting that the multi-component properties of SBJDD may help restore TME homeostasis and offer novel cancer therapy.

ObjectiveTo investigate the mechanism by which Shenbai Jiedu prescription （SBJDF） inhibits the proliferation of colorectal cancer （CRC） HCT116 cells. MethodAfter 48 h treatment of HCT116 cells with SBJDF （0， 0.25， 0.5， 1， 2， 4 g·L^-1）， the viability of HCT116 cells were determined by methyl thiazolyl tetrazolium （MTT） colorimetry. Following the classification of cells into blank control group and SBJDF （1， 2， 4 g·L^-1） groups， the effect of SBJDF on HCT116 cell morphology was observed under an inverted microscope. The effects of SBJDF on the proliferation of HCT116 cells and mitochondrial membrane potential （Δψm） were detected by colony formation assay and JC-1 probe， respectively. The flow cytometry was then performed for determining cell cycle distribution and apoptosis. The effects of SBJDF on cell cycle-， apoptosis-， and nuclear factor kappa-B （NF-κB） signaling pathway-related proteins were determined by Western blot. ResultSBJDF effectively inhibited the vitality of HCT116 cells and changed their morphology in a concentration-dependent manner. Compared with the blank control group， SBJDF at 1， 2， 4 g·L^-1 significantly reduced cell colony formation （P<0.05， P<0.01），and SBJDF at 2 and 4 g·L^-1 arrested the HCT116 cell cycle at G₀/G₁ phase （P<0.05， P<0.01）. Compared with the blank control group， SBJDF at 1， 2， 4 g·L^-1 remarkably down-regulated the protein expression of CyclinD₁ （P<0.05， P<0.01）. SBJDF at 2 and 4 g·L^-1 lowered the CyclinA₂ and cyclin-dependent kinase 4 （CDK4） （P<0.05， P<0.01）. SBJDF at 4 g·L^-1 reduced the cyclin-dependent kinase 1 （CDK1） （P<0.01）. Compared with the blank control group， SBJDF at 2 and 4 g·L^-1 induced HCT116 cell apoptosis， down-regulated the protein expression of anti-apoptosis-related proteins Bcl-2 and Bcl-xl as well as the NF-κB signaling pathway-related proteins IκB kinase α （IKKα），inhibitor α of NF-κB （IκBα），and phospho-NF-κB p65 （p-p65） （P<0.05， P<0.01）， and diminished the mitochondrial membrane potential of HCT116 cells. ConclusionSBJDF inhibits the proliferation of HCT116 cells， which may be related to its inhibition of the activation of NF-κB signaling pathway and the induction of cell cycle arrest and apoptosis.

ObjectiveTo study the mechanism of Shenbai Jiedu prescription inhibiting the proliferation of HCT116 colorectal cancer （CRC） cells by regulating the phosphatase and tensin homolog deleted on chromosome ten （PTEN）/phosphatidylinositol 3-kinase （PI3K）/ protein kinase B （Akt） signaling pathway. MethodShenbai Jiedu prescription was extracted by water extraction and alcohol precipitation to prepare freeze-dried powder. HCT116 cells were cultured in vitro， and treated with different concentrations of Shenbai Jiedu prescription （2， 4， 8， 16 g·L^-1）. The inhibitory effect of Shenbai Jiedu prescription on the proliferation of HCT116 cells was tested by methyl thiazolyl tetrazolium （MTT）. Real-time quantitative PCR was used to detect the mRNA expression levels of PTEN， PI3K， Akt， glycogen synthase kinase-3β （GSK-3β）， c-Myc， survivin and Cyclin D₁. Western blot was employed to measure the protein expression levels of PTEN， phosphorylated PTEN （p-PTEN）， PI3K， Akt， phosphorylated Akt （p-Akt）， GSK-3β， phosphorylated GSK-3β （p-GSK-3β）， c-Myc， survivin and Cyclin D₁， β-catenin nuclear import was explored by immunofluorescence assay. ResultCompared with the control group， Shenbai Jiedu prescription inhibited the proliferation of HCT116 cells in a dose-dependent manner （P<0.01）. Compared with the control group， the mRNA expression levels of PTEN and GSK-3β were up-regulated whereas those of PI3K， Akt， c-Myc， survivin and CyclinD₁ were down-regulated after treatment with Shenbai Jiedu prescription （P<0.01）. The protein expression levels of PTEN， p-PTEN and GSK-3β were up-regulated whereas those of PI3K， Akt， p-Akt， GSK-3β， p-GSK-3β， c-Myc， survivin and CyclinD₁ were down-regulated （P<0.05， P<0.01）. Immunofluorescence assay showed that Shenbai Jiedu prescription suppressed β-catenin nuclear import in HCT116 cells. ConclusionShenbai Jiedu prescription inhibited the proliferation of HCT116 cells via the mechanism of regulating the PTEN/PI3K/Akt signaling pathway.

Objective To analyze the infection rate of hepatitis B virus (HBV) in aggressive B-cell non-Hodgkin lymphoma (B-NHL), indolent B-NHL and multiple myeloma (MM) and its relationship with clinicopathological features. Methods The clinical data of 293 aggressive B-NHL, 181 indolent B-NHL and 261 MM patients in Tianjin Medical University Cancer Institute and Hospital from January 2009 to April 2017 were retrospectively analyzed. The difference of HBV infection was compared among three groups. Serum samples from all patients were tested for HBV markers, including hepatitis B surface antigen (HBsAg), hepatitis B surface antibody (HBsAb), hepatitis B e antigen (HBeAg), hepatitis B e antibody (HBeAb) and hepatitis B core antibody (HBcAb) by using chemiluminescence immunoassay. Results The positive rate of HBsAg was 9.2% (27/293), 5.5% (10/181) and 3.8% (10/261), respectively in the aggressive B-NHL group, indolent B-NHL group and MM group. The positive rate of HBsAg in the aggressive B-NHL group was higher than that in the indolent B-NHL group and MM group (χ2＝6.987, P＝0.030), and there was no statistical difference of HBsAg positive rate between the indolent B-NHL group and MM group (P ＞ 0.05). The positive rate of HBsAg, HBeAg, HBcAb in the aggressive B-NHL group was higher than that in the indolent B-NHL group and MM group [4.1% (12/293), 0, 0.8% (2/261); χ2＝ 14.976, P＝ 0.001], and there was no significant difference in the positive rate of HBsAg, HBeAg and HBcAb between the indolent B-NHL group and MM group (P ＞ 0.05). Compared with HBsAg negative aggressive B-NHL patients, HBsAg positive aggressive B-NHL patients showed, higher ratio of stage Ⅲ-Ⅳ [70.4% (19/27) vs. 49.2% (131/266), χ 2 ＝ 4.377, P＝0.036], more frequent involvement of spleen [51.9% (14/27) vs. 23.7% (63/266), χ 2＝ 10.039, P＝ 0.002], more frequent of B symptom [55.6% (15/27) vs. 32.0% (85/266), χ 2 ＝ 6.073, P＝ 0.014], more frequent of elevated total bilirubin [29.6% (8/27) vs. 14.3% (38/266), χ 2 ＝ 4.360, P ＝ 0.037] and more frequent of reduced albumin [55.6% (15/27) vs. 35.7% (95/266), χ 2＝ 4.115, P＝ 0.042]. Conclusions The infection rate of HBV in aggressive B-NHL patients is higher than that in the indolent B-NHL and MM patients. HBsAg positive aggressive B-NHL patients are associated with adverse clinical characteristics.

Objective To compare the effectiveness and mechanical differences in the dynamic hip screw (DHS), proximal femoral nail antirotation (PFNA), and proximal femoral internal fixator (PFI) for fixing complex unstable proximal intertrochanteric fractures by biomechanical testing. Methods Eighteen Synbones of the proximal femur were made to simulate complex unstable femoral intertrochanteric fracture models (Evans-Jensen TypeⅢ), which were fixed by DHS, PFNA, and PFI, respectively. The models were tested using a biomechanical testing machine, in order to compare their differences and advantages for fixing fractures. Results Under the compressive loads of 300, 600, and 1 200 N, the fracture displacement of the DHS was the maximum, with a significant difference compared with PFNA and PFI (P<0.05). There were no significant differences between PFNA and PFI (P>0.05). Under torsional loads of 300, 600, and 1 200 N, the torsional displacements of DHS and PFNA at the fracture ends were the maximum, with no significant difference (P>0.05). There were significant differences between PFI and PFNA as well as PFI and DHS (P<0.05). Conclusions For complex unstable proximal intertrochanteric fractures, the stability of the compression resistance of the PFI system is similar to that of the PFNA system. However, the torsional resistance of PFI is stronger than that of PFNA. The DHS system shows the least resistance with respect to compression and torsion.

Objective To study the nasal mucosa absorption of paeonol ofXingbi microemulsion-based thermo-sensitive gel in rats; To evaluate the nasal mucosa absorption of medicine and the rationality of preparation form.Methods According to the nasal administration method, the blood medicine concentration of paeonol was determined at different time points.Results The nasal administration bioavailability of paeonol in Xingbi microemulsion-based thermo-sensitive gel was 78.68%, the plasma concentration reached the maximum (0.3404 μg/mL) after 3 min of administration, and the average residence time was 9.23 h. After reaching Cmax, The plasma concentration changed similar to intravenous administration of paeonol. The concentration data of paeonol in plasma was consistent with a two-compartment model, with a weight of 1.Conclusion Xingbi microemulsion-based thermo- sensitive gel has a higher bioavailability, and the preparation form was reasonable.

Objective To establish the quality standards of Gualou Guizhi Dropping Pills; To investigate the vitro dissolution. Methods HPLC was used to determine the contents of paeoniflorin, albiflorin, cinnamic acid, 6-gingerol, liquiritin, glycyrrhizic acid and liquiritigenin in the dropping pills. The vitro dissolution rate of dropping pills was determined by rotating basket method. Results The calibration curves of paeoniflorin and albiflorin were in a good linearity in the range of 0.690–6.900 μg, 0.300–2.996 μg, respectively, and the average recoveries were 101.12% and 98.52%, respectively, with RSD of 2.24%, 1.37%, respectively. Cinnamic acid was linear in the range of 0.023–0.348 μg and the average recovery was 98.21% with RSD of 2.00%. 6-Gingerol was linear in the range of 0.025–0.382 μg, and the average recovery was 99.19% with RSD of 2.18%. The calibration curves of liquiritin, glycyrrhizic acid and liquiritigenin were in a good linearity in the ranges of 0.120–2.498 μg, 0.150–2.253 μg, and 0.010–0.147 μg, respectively, and the average recoveries were 99.80%, 100.38%, and 100.62%, respectively with RSD of 2.10%, 1.91%, 1.66%, respectively. The accumulated dissolution rates of seven kinds of elements in the dropping pills all reached more than 98% in 15 min. Conclusion The method is simple and accurate, with repeatability, which can be applied to the quality control of Gualou Guizhi Dropping Pills. The vitro dissolution rates meet relevant standards.

Objective To investigate the effects of Gualou Guizhi Granules on excitatory toxic damage of PC12 induced by glutamate; To primarily explore the involved protective mechanism of Gualou Guizhi Granules. Methods Excitatory toxic damage of PC12 induced by glutamate was used to establish models. Cells were divided into normal group, glutamate group, and Gualou Guizhi Granules low- (200 μg/mL), medium- (400 μg/mL) and high-dose (800 μg/mL) groups. MTT and LDH assay methods were used to detect PC12 activity; Caspase-3 activity detection method and Annexine V/PI double staining method were used to detect cell apoptosis; Western blot and RT-PCR were used to detect Bcl-2, Bax protein and mRNA expression. Results Compared with glutamate group, MTT showed that all Gualou Guizhi Granules groups could improve PC12 activity, and LDH showed that cell activity in all Gualou Guizhi Granules groups decreased; Annexine V/PI double staining method showed that all Gualou Guizhi Granules groups could decrease the PC12 apoptosis; Caspase-3 activity detection method showed that all Gualou Guizhi Granules groups could decrease the activity of Caspase-3; Western blot and RT-PCR showed that all Gualou Guizhi Granules groups could reduce Bax expression and increase Bcl-2 expression. Conclusion Gualou Guizhi Granules have certain protective effects on excitatory toxic damage of PC12 induced by glutamate, which may be related to its anti-apoptotic activity.

Objective T o establish a query table of IB S critical value and identification pow er for the detection system s w ith different num bers of ST R loci under different false judgm ent standards. Methods Sam ples of 267 pairs of full siblings and 360 pairs of unrelated individuals w ere collected and 19 auto-som al ST R loci w ere genotyped by G oldeneyeTM 20A system . T he full siblings w ere determ ined using IB S scoring m ethod according to the 'R egulation for biological full sibling testing'. T he critical values and identification pow er for the detection system s w ith different num bers of ST R loci under different false judgm ent standards w ere calculated by theoretical m ethods. Results A ccording to the form al IB S scoring criteria, the identification pow er of full siblings and unrelated individuals w as 0.7640 and the rate of false judgm ent w as 0. T he results of theoretical calculation w ere consistent w ith that of sam ple observation. T he query table of IB S critical value for identification of full sibling detection system s w ith different num bers of ST R loci w as successfully established. Conclusion T he IB S scoring m ethod defined by the regulation has high detection efficiency and low false judgm ent rate, w hich provides a relatively conservative result. T he query table of IB S critical value for identification of full sibling detection sys-tem s w ith different num bers of ST R loci provides an im portant reference data for the result judgm ent of full sibling testing and ow ns a considerable practical value.