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 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 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 evaluate the clinical outcomes of lumbopelvic fixation of vertically unstable Tile type C pelvic fractures combined with complex sacral fractures.Methods A prospective study was made on 21 cases of vertically unstable type C pelvic fractures combined with complex sacral fractures admitted between May 2014 and December 2015.There were 10 males and 11 females, with a mean age of 35.9 years (range, 14-59 years).Tile classification of pelvic fractures was type C1 in five cases, type C2 in four and type C3 in 12.Denis classification of sacral fractures was zone I in five cases, zone Ⅱ in seven and zone Ⅲ in nine.Twelve cases had neurological deficits.Operation time, intraoperative blood loss, bone healing time, pelvis vertical displacement and postoperative complications were recorded.Vertical displacement and functional outcome were assessed by Matta method and Majeed score respectively.Results All cases were followed up for (12.5±5.4)months (range, 7-26 months).Operation time was (108.0±49.4) min (range, 64-225 min).Intraoperative blood loss was 150-3 000 ml[400(225-500)ml].All fractures were healed at (19.0±4.6) weeks (range, 15-32 weeks).Vertical displacement of the pelvis was (8.76±5.46)mm (2.54-21.80 mm) before operation and (4.20±3.22)mm (0-12.57 mm) after lumbopelvic fixation (P<0.05), showing the reduction distance of-4.45-17.86 mm [4.09(1.74-5.58)mm].According to the Matta method, the results were excellent in 13 cases, good in six, and fair in two, with the excellent and good rate of 90%.Majeed score evaluation at last follow-up was 51-98 points and the results were excellent in nine cases, good in nine, fair in two and poor in one, with the excellent and good rate of 86%.Deep infection occurred in two cases who were cured by debridement and antibiotics, and the internal instruments of the two cases were removed after fracture healing.Screw loosening occurred in five cases with no evidence of screw breakage.Four cases complained of foreign body sensation and discomfort in sacral area due to the prominence of the iliac hardware.Four cases had limited range of motion of the lumbar spine, especially in anterior flexion range, and lumbar range of motion was recovered in three of them after removal of the internal fixator.Conclusion Lumbopelvic fixation can restore vertical stability of the pelvis, but removal of the internal fixator is suggested after fracture healing due to the high rate of screw loosening.

Objective To investigate the effect on incidence of ventilator associated pneumonia(VAP),the cost of hospitalization with closed endotracheal suctioning and vital sign as well in postoperative cardiac patients.Methods 304 postoperative cardiac patients supporting by ventilation were enrolled in this cohort study during January,2012-November,2013 in The Second affiliated Hospital& Yuying Children Hospital of Wenzhou Medical University.All the subjects were randomly divided into observational group and control group by coin side.Closed endotracheal suctioning system was applied in observation group and opened mode was applied in control group.Compare the vital sign(heart rate,blood pressure,saturation) at the moment of aspiration,suction time,incidence of VAP,duration of ventilation,mortality,the cost of suction,hospital stays and hospitalization expense.Results The baseline is no significant difference between two groups.The fluctuation of blood pressure and heart rate is lower in observational group at 30 second since completed the suction(P ＜0.05),but saturation is higher at 30 second and 60 second since completed the suction respectively(P ＜ 0.05).There is no significant difference of incidence of unexpected tube displacement and pneumothorax between two groups.Average time of each suction of experimental groups is shorter than Control groups[(156 ± 6) s vs (225 ± 8) s,t =-84.86,P ＜ 0.01].VAP incidence is lower in experimental group (12.0％ vs.18.6％,x2 =4.37,P ＜ 0.05).Duration of ventilation is lower in experimental group[(72 ± 33) h vs.(98 ± 38) h,t =-6.35,P ＜ 0.05].The cost of suction is higher in observational group [(346 ± 15) RMB vs.(178 ± 26) RMB,t =69.00,P ＜ 0.01],but the hospitalization expense is lower in experimental group [(32 011 ± 2 525) yuan vs.(35 264 ± 3 846)yuan,t =-8.72,P ＜ 0.05].There is no significant difference in mortality between two groups (x2 =0.08,P ＞ 0.05).Conclusion Application of closed endotracheal suction system can result in reduction vital sign fluctuation and incidence of cross infection and reducing the workload of nurses and decreasing the complication of suction,shorting the duration of ventilation and hospitalization and saving the expense of hospitalization in postoperative cardiac patients comparing with open mode.It is worthy to be populized in cardiac care unit.

Objective:To observe the expression of Ki-67,P53 and LAT1 in tissues of esophageal squamous cell carcinoma and precancerous lesions and to investigate its clinical significance.Methods: Immunohistochemical method was used to detect the expressions of Ki-67,P53 and LAT1 in tissues from 20 cases of normal esophageal mucosa,68 cases of precancerous lesions including 21 cases of mild atypical hyperplasia,22 cases of atypical hyperplasia,25 cases of severe atypical hyperplasia and cancer tissues from 34 esophageal cancer patients.The relationship between the expression of the three in esophageal carcinoma was analysed.Results: Re-spectively,the positive expression rate of Ki-67 in normal esophageal mucosa, mild atypical hyperplasia, moderate dysplasia, severe dysplasia and carcinoma was 0%( 0/20 ) , 23.8%( 5/21 ) , 40.9%( 9/22 ) , 76.0%( 19/25 ) , 82.4%( 28/34 ) and the positive expression rate of P53 was 0%(0/20),14.3%(3/21),31.8%(7/22),48.0%(12/25),67.6%(23/34) and the positive expression rate of LAT1 was 0%( 0/20 ) , 19.0%( 4/21 ) , 36.4%( 8/22 ) , 52.0%( 13/25 ) , 76.5%( 26/34 ).The rank correlation analysis showed that the positive expression of Ki-67,P53 and LAT1 were significantly correlated with histological grade(r=0.626,0.427, 0.586,P<0.01) and the expression of Ki-67,P53 was positively correlated with LAT1 in esophageal carcinoma tissues(r=0.428, 0.596,P<0.01).Conclusion:Abnormal expression of Ki-67,P53,LAT1 protein was significant related to carcinogenesis of esophageal cancer and the combined detection of the three has important clinical significance.