Objective To analyze the influence of usual processing steps on insoluble particles in Chinese medicine injection,with Shuanghuanglian injection as an example.Methods Light block method was adopted to analyze the influence of the different conditions of sterilization,ultra filtration and PH value on insoluble particles in Chinese medicine injection.Results Sterilization,ultra filtration and PH value caused the change of insoluble particles in Chinese medicine injection.Conclusion The comprehensive use of sterilization,ultra filtration,PH value can reduce the quantity of insoluble particles and clinical risk.

Objective:To investigate the effects of microRNA (miR)-513a-3p regulating hexokinase 2 (HK2) on proliferation and glycometabolism in colorectal cancer cell.Methods:From May 2019 to February 2020, the miR-513a-3p simulant, miR-513a-3p inhibitor and miR control were transfected into colorectal cancer SW480 cell respectively. Real-time quantitative polymerase chain reaction was used to detect the expression levels of miR-513a-3p in colorectal cancer SW480 cell, normal colorectal cell and all transfected colorectal cancer SW480 cell. The effect of miR-513a-3p on cell proliferation was detected by CCK-8 assay. Brd/PI incorporation assay was used to detect the effect of miR-513a-3p on glycometabolism. Western blot was used to detect the expression of HK2.Results:The expression level of miR-513a-3p in colorectal cancer SW480 cell was significantly lower than that in normal colorectal cell (0.43 ± 0.06 vs. 1.00 ± 0.02), and there was statistical difference ( t = 7.024, P = 0.003). The expression level of miR-513a-3p in colorectal cancer SW480 cell transfected with miR-513a-3p simulant was significantly higher than that in colorectal cancer SW480 cell transfected with miR control and transfected with miR-513a-3p inhibitor (1.18 ± 0.24 vs. 0.45 ± 0.04 and 0.22 ± 0.03), the expression level of miR-513a-3p in colorectal cancer SW480 cell transfected with miR control was significantly higher than that in colorectal cancer SW480 cell transfected with miR-513a-3p inhibitor, and there were statistical differences ( P<0.05). The proliferation ability in colorectal cancer SW480 cell transfected with miR-513a-3p simulant at 24, 48, 72 and 96 h was significantly lower than that in colorectal cancer SW480 cell transfected with miR-513a-3p control group, the proliferation ability in colorectal cancer SW480 cell transfected with miR-513a-3p inhibitor at 24, 48, 72 and 96 h was significantly higher than that in colorectal cancer SW480 cell transfected with miR-513a-3p control, and there were statistical differences ( P<0.05). The glucose intake, lactate and thioredoxin-interacting protein (TXNIP) expression levels in colorectal cancer SW480 cell transfected with stimulant were significantly lower than those in colorectal cancer SW480 cell transfected with miR control (1.02 ± 0.04 vs. 1.90 ± 0.06, 0.88 ± 0.03 vs. 1.45 ± 0.04 and 0.16 ± 0.02 vs. 0.86 ± 0.06), the indexes in colorectal cancer SW480 cell transfected with miR-513a-3p inhibitor were significantly higher than those in colorectal cancer SW480 cell transfected with miR control (2.35 ± 0.09 vs. 1.90 ± 0.06, 1.67 ± 0.08 vs. 1.45 ± 0.04 and 2.01 ± 0.15 vs. 0.86 ± 0.06), and there were statistical differences ( P<0.05). The expression level of HK2 in colorectal cancer SW480 cell transfected with miR-513a-3p stimulant was significantly lower than that in colorectal cancer SW480 cell transfected with miR control (0.20 ± 0.01 vs. 1.02 ± 0.04), and there was statistical difference ( t = 8.367, P<0.05); the expression level of HK2 in colorectal cancer SW480 cell transfected with miR-513a-3p inhibitor was significantly higher than that in colorectal cancer SW480 cell transfected with miR control (1.91 ± 0.07 vs. 1.02 ± 0.04), and there was statistical difference ( t = 4.279, P<0.05). Conclusions:MiR-513a-3p can significantly inhibit the proliferation and glycometabolism of colorectal cancer cell, and its regulatory mechanism is related to the inhibition of the HK2 protein expression in the cell by miR-513a-3p.

Objective:To investigate the relationship between the expression of inhibin subunit Beta A (INHBA) and the clinicopathological data and its effect on proliferation, invasion and metastasis of gastric cancer cells and its possible mechanisms.Methods:Using Gene Expression Profiling Interactive Analysis (GEPIA) public database to verify the expression of INHBA mRNA in gastric cancer and adjacent tissues and its relationship with pathological stage and prognosis; the relationship between INHBA and clinical prognosis was analyzed using the Kaplan-Meier Plotter online database. Immunohistochemistry was used to confirm the correlation between protein expression level of INHBA in gastric cancer and adjacent tissues and clinical pathological staging. In vitro, the cell proliferation was detected by tetrazolium salt (MTT) method; the cell migration ability was detected by scratch test, and cell invasion and metastasis ability was detected by transwell chamber assay. The expression of INHBA and epithelial-mesenchymal transition (EMT) related proteins was detected by Western blot. Results:The GEPIA database and Kaplan-Meier Plotter online database analysis showed that INHBA mRNA was highly expressed in various cancer tissues and significantly higher in gastric cancer tissues than normal tissues ( P<0.05). The high expression of INHBA mRNA was associated with clinical stage and poor prognosis of gastric cancer ( P<0.05). The results of immunohistochemistry showed that the staining score of INHBA in gastric cancer tissues was significantly higher than that in adjacent tissues ( P<0.05), and its expression level was correlated with clinical tumor node metastasis (TNM) stage ( P<0.05). The results of MTT, scratch test and transwell chamber showed that INHBA overexpression could promote the proliferation, migration, invasion and metastasis of gastric cancer cells, while interference with INHBA expression could inhibit the proliferation, migration, invasion and metastasis of gastric cancer cells; Western blot results showed that the expression of CDH1 was down regulated and the expression of CDH2 was up-regulated after INHBA overexpression. The expression of CDH1 was up-regulated and the expression of CDH2 was down-regulated after INHBA overexpression was inhibited. Conclusions:INHBA is highly expressed in gastric cancer tissues compared with adjacent tissues. The expression level of INHBA is related to tumor progression and poor prognosis. INHBA can promote the proliferation, migration and invasion of gastric cancer MGC-803 cell line and its mechanism may be related to INHBA promoting cell EMT.

Objective:To investigate the expression and clinical significance of growth differentiation factor 15 (GDF-15) in papillary thyroid carcinoma (PTC).Methods:The tumor tissues and metastatic lymph node tissues of 3 PTC patients who underwent radical surgery of thyroid cancer in the First Hospital of Hunan University of Chinese Medicine from January to February 2019 were collected, and the differential expressed genes were screened by high-throughput sequencing; 20 cases of primary tumor tissues and metastatic lymph node tissues were collected to verify the sequencing results. Another 20 cases of primary PTC tissues and adjacent tissues (>2 cm away from the tumor edge) were collected to verify the expression of target genes in tumor tissues and adjacent tissues. Sixty-four pathological specimens of PTC patients who underwent radical surgery of thyroid cancer in the First Hospital of Hunan University of Chinese Medicine from January to December 2014 were collected, of which 31 patients had lymph node metastasis. The real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of GDF-15 and verify the sequencing results; immunohistochemistry was used to detect the expression of GDF-15 protein in the primary PTC tissues, adjacent tissues and metastatic lymph node tissues. According to the expression of GDF-15 protein in the primary tumor tissues of PTC patients, the patients were divided into high-expression group (35 cases) and low-expression group (29 cases), and the relationship between GDF-15 expression level and clinicopathological characteristics of patients was analyzed; Kaplan-Meier method was used to analyze the 5-year tumor-free survival rate of the two groups.Results:The mRNA high-throughput sequencing results of 3 cases of PTC primary and metastatic tissues showed that the top 10 differential expressed genes were CDH2, CDF15, DKK1, GLIPR1, PCDH7, ID3, FBN1, MYPN, UBASH3B and CCDC80. The expression of GDF-15 mRNA in 20 cases of PTC primary tumor tissues and adjacent tissues were 4.1±0.5 and 2.8±0.3, and the difference was statistically significant ( t = 2.220, P = 0.032). The expression of GDF-15 mRNA in another 20 cases of PTC primary tumor tissues and metastatic lymph node tissues were 3.1±0.4 and 5.8±0.7, and the difference was statistically significant ( t = 3.556, P = 0.001). The results of immunohistochemistry showed that GDF-15 had the immunohistochemical scores of (4.0±0.3) points, (6.1±0.3) points and (9.0±0.4) points in PTC adjacent tissues, primary tumor tissues and metastasis tissues. The expressions of GDF-15 protein between PTC primary tumor tissues and adjacent tissues, metastatic tissues and adjacent tissues, and metastatic tissues and primary tumor tissues were significantly different (all P < 0.01). The differences in composition ratios of tumor long-axis diameter, tumor T stage and N stage between GDF-15 high-expression group and low-expression group were statistically significant (all P<0.05). The 5-year tumor-free survival rates in GDF-15 high-expression group and low-expression group were 60% and 83%, and the difference was statistically significant ( P = 0.033). Conclusions:The expressions of GDF-15 in PTC adjacent tissues, tumor tissues and metastatic lymph node tissues gradually increase, and its expression level is related to tumor progression, recurrence and metastasis. It can be used as a potential clinical prognostic warning molecule and therapeutic target.

Objective To establish a rabbit model of radiation-induced skeletal muscle injury in order to study the ultrastructural pathological changes and underlying mechanism.Methods 28 New Zealand rabbits were randomly divided into 2 groups with 16 rabbits in experimental group and 12 rabbits in control group.The experimental rabbits were irradiated on hip with a single dose of 80 Gy of 9 MeV electrons from a linear accelerator.1 month and 6 months after irradiation the pathological changes were respectively observed under light microscope and electron microscope.Results One month after irradiation,the morphologic changes including degeneration,necrosis of muscle cells,and hemorrhage between the muscle cells were observed under light microscope and the swelling of myofibrillae,blurring of light and shade band,vacuolar degeneration of mitochondria and amorphous areas of necrosis were observed under electron microscope.Six months after irradiation,the morphologic changes of nucleolus chips,fibrous connective tissue,thickening of vascular wall and vascular congestion between the muscle cells and the amorphous areas of necrosis in the experimental group were much more serious than those of 1 month after irradiation.In addition,the myofilaments were lost in degeneration areas and the sarcomere became shorten.Observation with electron microscope showed that the mitochondrial size and its morphological changes were varied and the amounts of collagen between myofibrillaes were increased 6 months after irradiation.Conclusions A rabbit model of high-dose irradiated skeleton muscle injury was successfully established with a single dose of 80 Gy of 9 MeV electrons from a linear accelerator.The degeneration and necrosis of muscle cells may be promoted by mitochondrial and vascular injury,degeneration of vessel and nerve fiber.

Objective To prospectively study the dynamic variation of vascular endothelial growth factor (VEGF),the short-term efficiency and the tolerability of the esophageal cancer patients treated by radiotherapy combined with thalidomide.Methods The serum samples of 86 esophageal cancer patients were collected before,during and after the radiotherapy.The VEGF levels were assayed by enzyme-linked immunosorbent assay (ELISA).3 patients interrupted the treatment because of intolerance radiotherapy.Based on the changes of VEGF level,32 esophageal cancer cases whose VEGF levels increased or remained were assigned to the group to which thalidomide was given during the whole course of radiotherapy.The rest 51 patients whose VEGF level decreased received radiotherapy without thalidomide during the whole course.In addition,30 healthy cases were included in control group.Then the efficiency and safety of the introduction of thalidomide in radiotherapy were investigated. Results The VEGF levels of 86 esophageal cancer cases were significantly higher than the 30 healthy control cases ( t =5.07,P ＜ 0.01 ),which were also correlated with the primary tumor size ( t =4.55,P ＜ 0.01 ),lymph node metastasis ( t =7.50,P ＜0.01 ),histological type( F =3.40,P ＜ 0.01 ) and clinical stage ( t =2.52,P ＜ 0.0 l ).However,it was irrelevant to the lesion site,distant metastasis,X-ray pathologic type,gender or age ( P ＞ 0.05).For the thalidomide involved group, the VEGF level after radiotherapy was significantly lower than during radiotherapy (t =2.37,P ＜0.05 ) with an effective rate of 71.88％.For the rest 51 cases without using thalidomide,the effective rate was 78.43％ yet there was no significant difference between the VEGF levels during and after radiotherapy ( t =0.18,P ＞ 0.05 ).62.50％ patients reported symptoms of dizzy and burnout after using thalidomide,while this incidence was 15.69％ for the rest patients (x2 =19.28,P =0.000).For the groups with or without thalidomide combination,the sleepiness incidences were 18.75％and 1.96％,respectively (x2 =5.168,P =0.023 ); the Ⅲ - Ⅳ grade esophagitis incidences were 12.50％ and 11.76％,respectively (x2 =0.061,P =0.806) ; the Ⅱ - Ⅳ grade leukocyte decrease incidences were 6.25％ and 9.80％,respectively (x2 =0.026,P =0.872) ; the Ⅲ - Ⅳ grade platelet descend incidences were 3.13％ and 5.88％,respectively (x2 =0.002,P =0.965 ) ; the Ⅲ - Ⅳ grade nausea and vomiting incidences were 9.38％ and 27.45％,respectively (x2 =2.913,P =0.088 ). No anaphylaxis was observed. Conclusions Thalidomide can decrease the VEGF expression level of esophageal cancer patients.Patients treated with thalidomide show good tolerance and compliance.

Objective To investigate the changes and clinical value of serum vascular endothelial growth factor (VEGF) level before,during and after radiotherapy in patients with esophageal carcinoma.Methods The sera of 67 esophageal carcinoma patients and 30 healthy control cases were collected.The VEGF level in serum samples were measured with enzyme-linked immunosorbent assay (ELISA) method.The relations among VEGF level changes,clinical stages and radiotherapy effect were analyzed.Results The VEGF levels of patients with esophagus cancer before,during and after radiotherapy were significantly higher than those in control group ( F =11.65,P ＜ 0.01 ).The VEGF level after radiotherapy was significant lower than that before radiotherapy ( F =10.72,P ＜ 0.01 ).The average VEGF level of patients with T3 and T4 was significantly higher than that of control group ( F =14.10,P ＜ 0.01 ).The average VEGF level of patients with N1 and N2 was significantly higher than that of control group( F =8.64,P ＜0.01).In 62 patients,the serum VEGF level increased in 21 cases but decreased in 41 cases after radiotherapy.With difference in radiotherapy efficiency of 61.90％ and 90.24％,respectively(x2 =6.08,P＜ 0.05).The average VEGF level during and after radiotherapy for 50 cases of CR + PR were significantly lower than that before radiotherapy( F =7.98,P ＜ 0.01 ).Conclusions Monitoring the serum VEGF level of patients with esophagus cancer can help evaluate the radiosensitivity,which has a significance in predicting the prognosis of radiotherapy.

Objective To explore the expression of COX-2,PD-ECGF and VEGF in gastric carcinoma tissue,and analyze and identify the relationship between the expression and tumor angiogenesis together with biological behaviour in gastric carcinoma.Methods ElivisionTM immunohistochemical method was performed to detect the expression of COX-2,PD-ECGF,VEGF and CD34,and evaluate the value of MVD in surgically resected gastric carcinoma specimens and biopsy tissues under gastroscope,including normal gastric mucosa tissues, metaplasia tissues and gastric mucosal atypical hyperplasia tissues. Clinicopathologic data of patients were analyzed retrospectively. Results The positive expression of COX-2, PD-ECGF,VEGF and MVD in gastric carcinoma were significantly higher than those in normal gastric mucosa, metaplasia or gastric mucosal atypical hyperplasia,and also their expression and MVD were closely related to lymph node metastasis and depth of invasion. The MVD in the groups of COX-2,PD-ECGF,VEGF positive-expression were significantly higher than those in negative-expression groups respectively. There were positive correlations among the expression of COX-2,PD-ECGF and VEGF in gastric carcinoma. MVD increased significantly when COX-2, PD-ECGF and VEGF coexpressed. Conclusion The expression of COX-2,PD-ECGF,VEGF and tumor angiogenesis participated in the genesis,invasion and metastasis of human gastric carcinoma.COX-2,PD-ECGF and VEGF participated in the tumor angiogenesis of gastric carcinoma. In the process of tumor angiogenesis and expression in gastric carcinoma, COX-2, PD-ECGF and VEGF could strengthen the effectiveness each other.They could strengthen the effective ness of tumor angiogenesis when they coexpressed. They could be selected as targets for tumor anti-angiogenesis treatment.

Objective:To observe the adverse reactions of killer cytokine-induced (CIK) in the treatment of malignant tumor and to analyze the possible mechanism ,and to develop the targeted prevention and treatment measures .Methods: The clinical data, including various adverse reactions , laboratory tests and the corresponding preventive measures against adverse reactions .In 1 240 patients with malignant tumor after treated with CIK cells from May 2013 to September 2015 were retrospectively analyzed .Results:The main adverse reactions after the first infusion of CIK cells were weak (10%),fever(7.25%),shiver (4%),arthralgia (3%),systemic in flammatory response syndrome reaction ( 3%) , digestive tract discomfort ( 0.96%) , acute allergic reaction ( 0.08%) , rash (0.08%),angina pectoris (0.08%),tumor lysis syndrome(0%),infection(0%).With the increase of the treatment ,the incidence of adverse reactions increased and the fever was the main performance ,after the fourth course into the platform .The combination of blood pressure increased or decreased and severe allergic reaction and systemic inflammatory response syndrome was needed to be treated .The CIK cells were pretreated before treatment could reduce the incidence of these reactions .Conclusion:CIK cells therapy is a safe and effective adoptive immunotherapy for malignant tumor and its adverse reactions can be treated expectantly , but rare adverse reactions may have potential risks .

Objective To investigate the applicability of zebra fish thrombosis model in antithrombotic activity screening of Chinese materia medica.Methods The living zebra fish thrombosis model was induced by adrenaline hydrochloride. Zebra fish were randomly divided into blank control group, model group, positive medicine group and medication group. Each group was given the corresponding medicine or embryo culture water. O-anisidine staining solution was used to stain and calculate the staining intensity of erythrocytes in zebra fish heart, and quantitative analysis was carried out. The platelet aggregation of transgenic zebra fish was observed and under qualitative analysis. Results Compared with the model group, 100μg/mL salvianolic acid B, 300, 900μg/mL aqueous extract of Salvia miltiorrhiza, 45μg/mL 95% ethanol extract and 400, 1200μg/mL hypothalamus could significantly inhibited the formation of zebra fish thrombosis (P<0.01).ConclusionZebra fish thrombosis model has good applicability in antithrombotic activity screening of Chinese materia medica.