ObjectiveTo investigate the molecular mechanisms by which salvianolic acid B （SalB） inhibits epithelial-mesenchymal transition （EMT） in non-small cell lung cancer （NSCLC） by downregulating phosphoribosylaminoimidazole carboxylase and phosphoribosylaminoimidazole succinocarboxamide synthetase （PAICS） expression. MethodsNSCLC A549 cells and normal bronchial epithelial cells （bronchial epithelium transformed with Ad12-SV40 2B， BEAS-2B） were used as models. Cell viability was assessed using the cell counting kit-8 （CCK-8） assay after treatment with SalB （0， 50， 100， 200， 300， 400， 500 μmol·L^-1 for 24 or 48 h to determine effective and safe intervention concentrations. Cell proliferation， cell cycle distribution， and apoptosis were evaluated by 5-ethynyl-2′-deoxyuridine （EdU） staining and flow cytometry， respectively. Wound healing and Transwell invasion assays were performed to assess cell migration and invasion. RNA sequencing combined with bioinformatic analysis was conducted to identify differentially expressed genes and functional enrichment. Molecular docking was used to predict the binding ability between SalB and PAICS， and the cellular thermal shift assay （CETSA） was performed to evaluate the effect of SalB on the thermal stability of the PAICS protein. Western blot （WB） was used to detect the effects of SalB on PAICS and EMT-related proteins （E-cadherin， N-cadherin， Vimentin， Snail， and Slug）. A functional rescue assay was conducted by PAICS overexpression via plasmid transfection. ResultsCompared with the control group， SalB inhibited A549 cell viability in a dose-dependent manner （P<0.05）， and the effective concentrations （≤300 μmol·L^-1） showed no significant cytotoxicity in BEAS-2B cells. Within this concentration range， SalB significantly inhibited A549 cell proliferation， migration， and invasion， and induced G₀/G₁ phase arrest and apoptosis （P<0.05）. Transcriptomic analysis showed that SalB significantly downregulated PAICS expression， and its functions were enriched in cell proliferation and EMT. Bioinformatic analysis indicated that PAICS is highly expressed in lung adenocarcinoma and is associated with poor prognosis （P<0.01）. Molecular docking showed that SalB has strong binding ability to PAICS （binding energy -9.1 kcal·mol^-1. CETSA results showed that SalB significantly increased the thermal stability of the PAICS protein （P<0.05）. WB results showed that， compared with the control group， SalB dose-dependently downregulated PAICS expression， upregulated E-cadherin， and downregulated N-cadherin， Vimentin， Snail， and Slug （P<0.05）. Functional rescue experiments showed that， compared with the empty vector group， PAICS overexpression significantly enhanced A549 cell proliferation， migration， and invasion， promoted cell cycle progression， and inhibited apoptosis （P<0.05）. Meanwhile， compared with the empty vector + SalB-H group， PAICS overexpression partially reversed the inhibitory effects of SalB on malignant phenotypes and EMT-related proteins （N-cadherin， Vimentin， Snail， and Slug）， and downregulated E-cadherin expression （P<0.05，P<0.01）， indicating that PAICS is a key functional target mediating the antitumor effects of SalB. ConclusionSalB effectively inhibits EMT progression and cell cycle progression in A549 cells by downregulating PAICS expression， thereby exerting anti-NSCLC effects. This study not only reveals that PAICS is a key functional target through which SalB regulates EMT， but also provides experimental evidence supporting SalB as a potential candidate drug for inhibiting NSCLC metastasis.

ObjectiveTo investigate the molecular mechanisms by which salvianolic acid B （SalB） inhibits epithelial-mesenchymal transition （EMT） in non-small cell lung cancer （NSCLC） by downregulating phosphoribosylaminoimidazole carboxylase and phosphoribosylaminoimidazole succinocarboxamide synthetase （PAICS） expression. MethodsNSCLC A549 cells and normal bronchial epithelial cells （bronchial epithelium transformed with Ad12-SV40 2B， BEAS-2B） were used as models. Cell viability was assessed using the cell counting kit-8 （CCK-8） assay after treatment with SalB （0， 50， 100， 200， 300， 400， 500 μmol·L^-1 for 24 or 48 h to determine effective and safe intervention concentrations. Cell proliferation， cell cycle distribution， and apoptosis were evaluated by 5-ethynyl-2′-deoxyuridine （EdU） staining and flow cytometry， respectively. Wound healing and Transwell invasion assays were performed to assess cell migration and invasion. RNA sequencing combined with bioinformatic analysis was conducted to identify differentially expressed genes and functional enrichment. Molecular docking was used to predict the binding ability between SalB and PAICS， and the cellular thermal shift assay （CETSA） was performed to evaluate the effect of SalB on the thermal stability of the PAICS protein. Western blot （WB） was used to detect the effects of SalB on PAICS and EMT-related proteins （E-cadherin， N-cadherin， Vimentin， Snail， and Slug）. A functional rescue assay was conducted by PAICS overexpression via plasmid transfection. ResultsCompared with the control group， SalB inhibited A549 cell viability in a dose-dependent manner （P<0.05）， and the effective concentrations （≤300 μmol·L^-1） showed no significant cytotoxicity in BEAS-2B cells. Within this concentration range， SalB significantly inhibited A549 cell proliferation， migration， and invasion， and induced G₀/G₁ phase arrest and apoptosis （P<0.05）. Transcriptomic analysis showed that SalB significantly downregulated PAICS expression， and its functions were enriched in cell proliferation and EMT. Bioinformatic analysis indicated that PAICS is highly expressed in lung adenocarcinoma and is associated with poor prognosis （P<0.01）. Molecular docking showed that SalB has strong binding ability to PAICS （binding energy -9.1 kcal·mol^-1. CETSA results showed that SalB significantly increased the thermal stability of the PAICS protein （P<0.05）. WB results showed that， compared with the control group， SalB dose-dependently downregulated PAICS expression， upregulated E-cadherin， and downregulated N-cadherin， Vimentin， Snail， and Slug （P<0.05）. Functional rescue experiments showed that， compared with the empty vector group， PAICS overexpression significantly enhanced A549 cell proliferation， migration， and invasion， promoted cell cycle progression， and inhibited apoptosis （P<0.05）. Meanwhile， compared with the empty vector + SalB-H group， PAICS overexpression partially reversed the inhibitory effects of SalB on malignant phenotypes and EMT-related proteins （N-cadherin， Vimentin， Snail， and Slug）， and downregulated E-cadherin expression （P<0.05，P<0.01）， indicating that PAICS is a key functional target mediating the antitumor effects of SalB. ConclusionSalB effectively inhibits EMT progression and cell cycle progression in A549 cells by downregulating PAICS expression， thereby exerting anti-NSCLC effects. This study not only reveals that PAICS is a key functional target through which SalB regulates EMT， but also provides experimental evidence supporting SalB as a potential candidate drug for inhibiting NSCLC metastasis.

OBJECTIVE To optimize the forming process of Kaixin granules and establish its physical fingerprint.METHODS Using particle forming rate， hygroscopicity rate， dissolution rate， and angle of repose as indices， the key process parameters of the granule forming process were determined by Plackett-Burman design. The analytic hierarchy process （AHP） combined with the entropy weight method was used to comprehensively weight the indices， and the forming process of Kaixin granules was optimized and validated by Box-Behnken response surface methodology. A physical fingerprint of Kaixin granules was established based on the following physical indices： angle of repose， Hausner ratio， bulk density， tapped density， moisture content， hygroscopicity ratio， and relative uniformity index， with similarity evaluation was performed. Cluster analysis and partial least squares discriminant analysis （PLS-DA） were performed using SPSS 20.0 and the SPSSPRO online analysis platform， respectively.RESULTS The key process parameters were ethanol volume fraction， ethanol amount， and the drug-excipient ratio. The optimal forming conditions were as follows： ethanol volume fraction of 87%， ethanol amount of 27%， and a drug-excipient ratio of 1∶1. Validation showed that the average comprehensive scores of three batches were 97.82， with an RSD of 0.19%. The similarity of the physical fingerprints of ten batches of Kaixin granules prepared under the optimal forming process were greater than 0.98. The results of cluster analysis showed that the ten batches were divided into two groups： S4， S5， S7， S8 and S10 in one cluster， and S1-S3， S6 and S9 in another. The variable importance in projection values of Hausner ratio， relative uniformity index， bulk density， and moisture content were greater than 1.CONCLUSIONS The optimized forming process is stable and feasible. The established physical fingerprint can effectively characterize the overall physical properties of Kaixin granules. Hausner ratio， relative uniformity index， bulk density， and moisture content are the key physical indices.

OBJECTIVE To optimize the water extraction technology for Kaixin granules. METHODS UPLC-MS/MS method was established for the simultaneous determination of ginsenoside Rg1， ginsenoside Re， ginsenoside Rb1， tenuifolin， polygalaxanthone Ⅲ and 3， 6′-disinapoyl-sucrose. An orthogonal test was designed with extraction times， extraction duration， and the volume of added water as factors. Using the contents of the aforementioned six indicator components and the extract yield as evaluation indexes， analytic hierarchy process-entropy weight method was employed to determine the combined weights of each indicator. Subsequently， process optimization and validation were conducted by integrating grey relational analysis （GRA） and back propagation （BP） neural network. RESULTS The water extraction technology optimized by the orthogonal test and GRA was 10- fold water for the first decoction and 8-fold water for the subsequent two， extracting 3 times，extracting for 1 h each time； the average comprehensive score of the validation experiment was 91.10 （RSD＝0.31%， n＝3）. The water extraction technology optimized by BP neural network was extracting 3 times with 10-fold water added each time， extracting for 1.5 h each time； the average comprehensive score of the validation experiment was 95.89 （RSD＝0.73%， n＝3）. Considering practical production requirements， the optimal water extraction technology was extraction performed three times， with 10-fold water for the first decoction and 8-fold water for the subsequent two extractions， with an extraction time of 1 h each. CONCLUSIONS The optimized water extraction technology for Kaixin granules is stable and feasible.

Objective:To evaluate the role of heme oxygenase-1 (HO-1) in hemopexin (HPX)-induced reduction of cerebral ischemia-reperfusion (I/R) injury in rats.Methods:One hundred and eighty male Sprague-Dawley rats, aged 7-8 weeks, weighing 250-280 g, were divided into 5 groups ( n=36 each) using a random number table method: sham operation group (group SH), I/R group, vehicle group (group V), HPX group, and HPX plus HO-1 specific inhibitor ZnPPIX(HZ group). Local cerebral I/R was produced by middle cerebral artery occlusion for 120 min followed by reperfusion in anesthetized rats.In I/R, V, HPX and HZ groups, 0.9% normal saline 10 μl, 0.1% NaN 3 10 μl, 1.86 g/L HPX (diluted to 10 μl in 0.1% NaN 3 solution), and 1.86 g/L+ ZnPPIX 20 μmol/L (diluted to 10 μl in 0.1% NaN 3 solution) were injected through the lateral ventricle, respectively, immediately after onset of reperfusion.Morris water maze test was used to detect the cognitive function on day 1 before ischemia and day 2 of reperfusion.Rats were sacrificed, brains were removed and brain tissues were obtained for determination of the permeability ratio of Evans blue (EB), brain water content, expression of VE-cadherin in ischemic penumbra (by Western blot), and expression of angiopoietin-1 (Ang1) mRNA and Ang2 mRNA (by real-time polymerase chain reaction). Ang1 mRNA/Ang2 mRNA ratio was calculated.CD31/vWF double labeling immunofluorescence was used to detect the density of neovascularization in hippocampal tissues in the ischemic penumbra. Results:Compared with SH group, the escape latency was significantly prolonged at 2-7 days of reperfusion, the percentage of time of staying at the target quadrant was decreased, the frequency of crossing the platform was decreased, the permeability ratio of EB in brain tissues was increased at day 7 of reperfusion, and the brain water content, Ang1 mRNA/Ang2 mRNA ratio, expression of VE-cadherin and density of neovascularization were decreased in I/R, V, HPX and HZ groups ( P<0.05). Compared with I/R group, the escape latency was significantly shortened at 2-7 days of reperfusion, the percentage of time of staying at the target quadrant was increased, the frequency of crossing the platform was increased, the permeability ratio of EB in brain tissues was decreased at day 7 of reperfusion, and the brain water content, Ang1 mRNA/Ang2 mRNA ratio, expression of VE-cadherin and density of neovascularization were increased in HPX group ( P<0.05), and no significant change was found in the parameters mentioned above in V and HZ groups ( P>0.05). Compared with HPX group, the escape latency was significantly prolonged at 2-7 days of reperfusion, the percentage of time of staying at the target quadrant was decreased, the frequency of crossing the platform was decreased, the permeability ratio of EB in brain tissues was increased at day 7 of reperfusion, and the brain water content, Ang1 mRNA/Ang2 mRNA ratio, expression of VE-cadherin and density of neovascularization were decreased in HZ group ( P<0.05). Conclusion:HO-1 is involved in HPX-induced reduction of cerebral I/R injury in rats.

Intraoperative radiotherapy (IORT) is a radiotherapy method that uses a radiotherapy device to provide a brachytherapy with single high-dose radiation to a target tissue (primary tumor beds,residual tumors,and lymphatic drainage areas) during surgery,which includes intraoperative electron beam radiotherapy and high-dose-rate intraluminal brachytherapy.Nowadays,the implementation of precise radiation dose is an important part of multidisciplinary,individualized treatment of tumors.IORT achieves precise treatment by maximizing the radiation dose to the target volume and protecting surrounding normal tissues as much as possible.This article describes the recent research on IORT.

Objective To preliminarily explore the delineation of clinical target volume in intensity-modulated radiotherapy for para-aortic lymph node metastases in patients with gynecological malignancies.Methods A retrospective analysis was performed on 56 gynecological tumor patients with para-aortic lymph node metastases who were admitted to our department from January 2010 to September 2016.The number and distribution of metastatic para-aortic lymph nodes were determined by imaging method.Results A total of 108 positive para-aortic lymph nodes were found in the 56 patients,with 1-4(mean,2) positive para-aortic lymph nodes per patient.The mean diameter of positive para-aortic lymph nodes was 2.3 cm (1.2-4.0 cm).A total of 20 metastatic lymph nodes (19%) were located at the L4 level,38(35%) at the L3 level,44(41%) at the L2 level,and 6(5%) at the L1 level.There were 71 metastatic lymph nodes (66%) at the left side of the para-aortic region,20 metastatic lymph nodes (19%) between the abdominal aorta and the vena cava,and 17 metastatic lymph nodes (15%) at the right side of the inferior vena cava.Conclusions For patients with gynecological malignancies,nodal contouring for the para-aortic region should not be defined by a fixed circumferential margin around the vessels.The left side of the para-aortic region should be covered adequately;the upper target should be extended up to the renal artery,and needs to be further extended for patients who have nodal involvement near the renal arteries and veins.

Objective:To investigate the effect of antlerbase on the radiation carcinogenesis by observing the effect of antlerbase on the expressions of Notch2 gene and immune function of the mice with thymic lymphomas induced by ionizing radiation.Methods:Ninety BALB/c mice were randomly divided into control group,irradiation group and irradiation combined with drug group (combined group)(n=30).The thymic lymphoma models of mice were made by X-ray irradiation in irradiation group and combined group;after establishing the animal models,the mice in combined group were fed with chow containing antlerbase superfine powder.After 6 months,the whole blood and thymic tissue were taken,then RT-PCR and Western blotting methods were used to detect the mRNA and protein expressions levels of Notch2 gene in the thymic lymphoma tissue;ELISA method was used to detect the serum immunoglobulin (IgG,IgA,IgM)levels and SOD kit was used to detect the serum SOD activities of the tumor mice.Results:The incidences of thymic lymphoma in irradiation group and combined group were 53.33%(16/30)and 36.67% (11/30),respectively.Compared with control group,the expression levels of mRNA and protein of Notch2 gene in thymic lymphoma of the mice in irradiation group were significantly increased (P <0.05);compared with irradiation group,the expression levels of mRNA and protein of Notch2 gene in combined group were decreased (P <0.05);compared with control group,the levels of IgG,IgA,and IgM in serum of the mice in irradiation group were decreased (P < 0.05 ); compared with irradiation group, the levels of IgG, IgM, hemoglobin and the activity of SOD in serum of the mice in combined group were increased (P < 0.05 ). Conclusion:Ionizing radiation can activate the high expression of Notch2 gene and protein and decrease the immune function of mice,it might be one of the mechanisms for the occurrence of radiation carcinogenesis;antlerbase can decrease the incidence of radiation carcinogenesis by inhibiting the expression of Notch2 gene and increasing the immune function of mice,which might play an role in restraining tumor.

Objective To explore the potential effects of endothelial progenitor cells (EPCs)-angiogenesis on mechanism of alleviating cognitive dysfunction in rats subjected to cerebral ischemia-reperfusion (I/R) injury. Methods A total of 121 male Sprague–Dawley (SD) rats were randomly divided into four groups:Sham group (n=31), focal I/R(MCAO, 0.9%saline 10μL, n=30) group, MCAO+Vehicle (sodium azide, 0.1%Vehicle 10μL, n=30) group and MCAO+HPX (1.86 g/L HPX 10μL, n=30) group. The modified neurological severity scores (mNSS) was carried out to determine neurological function deficit after I/R. Morris water maze (MWM) was carried out to assess learning and memory abilities after I/R. The circulating EPCs after I/R were counted by flowcytometry (FCM) combined with double-immunofluorescence staining of CD34 and CD133. Angiogenesis in rat penumbra cortex after I/R was assessed by immunohistochemical technique combined with immunofluorescent chromogenic detection of CD31 and vWF. Results Compared with sham group, the mNSS scores, the escape latency and the circulating EPCs count were increased after I/R, the time percentage spent in target quadrant was reduced, and the new vessel density in penumbra cortex was increased after I/R in MCAO group (P < 0.05 respectively). There were no significant differences in mNSS score, the escape latency, the time percentage spent in target quadrant, the circulating EPCs count and the new vessel density in penumbra cortex between MCAO group and MCAO+Vehicle group ( P>0.05). The mNSS score and the escape latency were significantly decreased, the circulating EPCs count and new vessel density in penumbra cortex were significantly increased after I/R in MCAO+HPX group compared with those of MCAO+Vehicle and MCAO group (P<0.05). Conclusion EPCs-angiogenesis signaling plays positive effects on HPX alleviating cognitive dysfunction in rats subjected to focal cerebral ischemia reperfusion injury.

OBJECTIVE: To observe the curative effects of haemocoagulase on patients with reduced coagulation after the transurethraal plasmakentic vaporization of prostrate(TUPKVP).METHODS: 132 cases were randomly divided into two groups,one treated with haemocoagulase,and the other as control group.Then two groups were compared for the time of operation,quantity of blood transfusion during the operation and the time difference in continuous bladder-washing after the operation.RESULTS: There was an obvious difference(P