OBJECTIVE To evaluate the inhibitory effect and possibility inhibitory mechanisms ofendostatin on laryngeal carcinoma cells (Hep-2) and human umbilical vein endothelial cells (HUVEC). METHODS ①Using the MTT assay , the effect of different concentrations of endostatin (10 ?g/ml～50 ?g/ml) and endostatin (30 ?g/ml) with different contact times (24～72h) on the livability of Hep-2 and HUVEC cells were determined; ②Changes in the ultrastructures of Hep-2 and HUVEC cells were examined by electron microscopy; ③Survivin mRNA content was determined in Hep-2 cells exposed to endostatin (30 ?g/ml) by RT-PCR test; ④The effect of endostatin (30 ?g/ml) on the ectogenetic artificial blood vessel models was observed by light microscopy. RESULTS ① The growth of Hep-2 and HUVEC cells was significantly inhibited (P

Objective To explore the influence of alpha-fetoprotein (AFP) on the expression of drug-resistance gene MDR1 and chemotherapeutic sensitivity in hepatocellular carcinoma (HCC) cells.Methods A HCC cell line SMMC-7721/AFP, which was stably transfected with AFP gene, was established.mRNA and protein expressions of AFP and MDR1 were detected by real-time PCR and Western Blot,respectively.The sensitivity of SMMC-7721/AFP and SMMC-7721/EGFP cells with or without MDR1 silencing by siRNA to doxorubicin was tested by MTT assay.Immunohistochemistry was used to detect the expression of MDR1 genes-coded protein Pgp in 60 cases of HCC tissues, and the relationship between Pgp expression and serum AFP levels was analyzed.Results AFP mRNA and protein could be detected in SMMC-7721/AFP cells, but not in control cells, indicating that the AFP stably transfected cell line was successfully established.MDR1 mRNA and protein levels were higher in SMMC-7721/AFP cells than those in SMMC-7721/EGFP cells.MDR1 mRNA level in SMMC-7721/AFP cells was (52.7 ± 1.5) times as high as that in SMMC-7721/EGFP cells (P ＜ 0.05).The resistance to doxorubicin was increased by (12.8 ± 1.1) times after AFP transfection (P ＜ 0.05).The chemosensitivity to doxorubicin was increased after the expression of MDR1 was knocked down by siRNA.The expression of Pgp in HCC tissues was positively correlated with the serum AFP levels.Conclusion AFP could induce drug-resistance to doxorubicin in HCC cells by increasing the expression of MDR1.

Objective To explore the clinical features, surgical techniques and treatment efficacy of blood blis?ter-like aneurysms (BBA) of the internal carotid artery (ICA). Method We retrospectively reviewed 45 BBAs of the ICA treated surgically at our department between Jan. 2006 and Jan. 2016. The data was analyzed. Results Direct clipping of BBA was performed in 38 patients, clipping over wrapping in 3 patients and trapping of ICA in 4 patients. Intraoperative rupture occurred in 12 patients (27%). Postoperative cerebral infarction developed in 10 cases (22%). Seven patients died (16%). During the follow-up time of 47 months (range, 6~126 months), Good results were achieved in 34 patients (76%) with a modified Rankin Scale (mRS) 0~2 and 4 patients were in mRS 3~5. Follow-up angiography revealed a re?currence of BBA in one patient, which was retreated with endovascular coiling. No rebleeding was found in all fol?lowed-up patients. Conclusions BBAs of the ICA are difficult to treat surgically with high risk and complications. Thoughtful pre-operative evaluation of BBA and application of various surgical strategies during operation can signifi?cantly reduce the surgical risk and achieve good outcomes.

Child ; Epiphyses ; Fracture Fixation, Internal ; Humans ; Lower Extremity

In order to investigate the prevalence and track genetic and antigenic evolutions of infectious bronchitis virus (IBV) and their prevalence in Guangxi, China since 1985, gene amplification and sequencing and virus neutralization (VN) test on chicken embryo tracheal organ cultures were used in genotyping and serotyping of 28 IBV isolates during 2009-2011 in Guangxi. The results of N gene sequencing and comparison showed that the 28 isolates and reference strains were classified into three groups, and most isolates belonged to group Ill, while the isolates in 1985-2008 belonged to groups IV and II. The data of VN test indicated that the 28 isolates belonged to 6 serotypes; among them, 71. 4% belonged to serotypes 1, 2, and 3, and 11 (39.3%) shared the same serotype with the current vaccine strains. Given the data of our previous study, it is found that prevalent serotypes and their proportions varied in different areas of Guangxi and during different periods. These data lay a good foundation for developing an oil-emulsified inactivated polyvalent vaccine containing local dominant serotypes for the effective prevention and control of infectious bronchitis.
Animals ; Antibodies, Viral ; immunology ; Chick Embryo ; Chickens ; China ; epidemiology ; Coronavirus Infections ; epidemiology ; immunology ; veterinary ; virology ; Infectious bronchitis virus ; classification ; genetics ; immunology ; isolation & purification ; Molecular Sequence Data ; Phylogeny ; Poultry Diseases ; epidemiology ; immunology ; virology

To explore the influence of matrix materials in complicate ingredients on traditional Chinese medicine and investigate the excipients selection model based on balanced release characteristics of multicomponents, the influence of HPMC (K4M, K15M, K100M) and Carbomer (934P, 971P, 974P) was illustrated by testing in vitro release of ginsenoside-Rg1, ginsenoside-Rb1 and notoginsenoside-R1 in Panax notoginseng saponins (model drug, PNS). According to in vitro release results of PNS matrix tablets in water and artificial intestinal juice, the release curves were analyzed with Peppas equation and simulating factor (f). Significant differences in k value and n value among ginsenoside-Rg1, ginsenoside-Rb1 and notoginsenoside-R1 existed in various formulations. The release behaviors from various excipients could be described with Non-Fickian transport or super Case II transport pattern. The f2 values for ginsenoside-Rg1, ginsenoside-Rb1 and notoginsenoside-R1 in 971P matrix tablet containing 30% Carbomer 971P were 74.91, 53.45, 57.89 in water and 79.35, 55.51, 51.89 in artificial intestinal juice, respectively. The release profiles fit for the regulation of FDA. The result revealed that the balanced release rates of Rg1, Rb1 and R1 in 971P matrix tablet were obtained.
Acrylates ; chemistry ; Acrylic Resins ; chemistry ; Delayed-Action Preparations ; Excipients ; chemistry ; Ginsenosides ; isolation & purification ; pharmacokinetics ; Lactose ; analogs & derivatives ; chemistry ; Methylcellulose ; analogs & derivatives ; chemistry ; Panax notoginseng ; chemistry ; Plants, Medicinal ; chemistry ; Saponins ; administration & dosage ; isolation & purification ; pharmacokinetics ; Tablets

Objective To prepare recombinant adenovirus pAd-gal-9 containing murine galectin-9 and explore galectin-9's pro-apoptotic effect on T lymphocytes. Methods The recombinant adenovirus plas-mid pAd/CMV/V5-DEST-gal-9 was prepared by conventional molecular cloning and LR reaction. The pAd/ CMV/V5-DEST-gal-9 linearlized by Pac I was transfected into 293A cells with Lipofectin 2000. Eight days after transfection, the 293A cells were subjected to freeze/thraw circle for three times and the supernatant was collected after centrifugation. Higer titer pAd-gal-9 was produced by large-scale infection of 293A cells with the supernatant containing pAd-gal-9. The supernatant was condensed to get purified pAd-gal-9 by CsCl density gradient centrifugation. After titer determination with gradient dilution of harvested pAd-gal-9 infec-tion in 293A-seeded 96-wells, pAd-gal-9 was used to infect the CHO cell line. Immunohistological assay, Western blot and flow cytometry were employed to ascertain the subcellular location expression of galectin-9. We added solid-phase transgenic CHO cells or freshly-cultured supernatant to medium containing activated T cells to detect the pro-apoptotic effect of galectin-9. Results The pAd-gal-9 was prepared successful. Im-munohistochemical staining of CHO infected with pAd-gal-9 confirmed that galectin-9 was expressed in the cytosol. Intercellular staining indicated that mean fluorescence intensity of galectin-9 was significantly higher in pAd-gal-9-infected CHO group than control group. Supernatant from pAd-gal-9-infected CHO promoted the apoptosis of T cells. The percent of apoptotic T cells was higher than the Tim-3 positive T cells. Conclu-sion CHO infected with pAd-gal-9 can secret galectin-9 to promote the apoptosis of activated T cells via Tim-3-independent mechanisms.

Objective: To explore the clinical characteristics, therapeutic methods and effects of blood blister-like aneurysms (BBA) of the supraclinoid segment of the internal carotid artery (ICA). Methods: A total of 40 patients with BBAs of the ICA treated at Department of Neurosurgery of West China Hospital between January 2010 and January 2016 were retrospectively reviewed. There were 14 males and 26 females with the mean age of 44.7 years (ranging from 25 to 64 years). There were 38 patients presented with subarachnoid hemorrhage (SAH), and 2 patients were found incidentally with unruptured aneurysms in regular examination. Surgical treatment was performed in 22 patients, including direct clipping, clipping over wrapping and trapping. Endovascular treatment with single or multiple stents and coiling was performed in 18 patients. Clinical outcomes were assessed with modified Rankin Scale. The patients were followed up with CT angiography. Results: Direct clipping of BBA was performed in 18 patients, clipping over wrapping in 3 patients and trapping of ICA in 1 patient. Intraoperative rupture occurred in 5 patients. Postoperative cerebral infarction developed in 6 cases, 4 patients died. Single stent-assisted coiling was performed in 8 cases, and multiple stents with coiling in 10 patients. Procedure-related hemorrhage occurred in 2 patients, postoperative massive cerebral infarction developed in 1 patient with stent-assisted coiling. The follow-up time was 26.8 months (ranging from 6 to 72 months), among the patients with surgery, good results (modified Rankin Scale 0-2) were achieved in 16 patients; one patient was score 3, and the other was 4. In endovascular group, 15 patients had good results and one with mRS 4. Follow-up angiography revealed recurrence of BBA in 2 patients treated with stent and coiling, which was retreated with coiling. No recurrence was found in surgical group. Conclusions BBA of the supraclinoid ICA is a specific and complicated intracranial aneurysm with high risk and complications no matter the treatment is surgical or endovascular, take proper operation methods could achieve satisfactory results.

Objective To investigate the effects of different extracts of pericarpium citri reticulatae (PCR) and pogostemon cablin benth (PCB) on the contraction of gastrointestinal smooth muscle and the level of gastrointestinal hormones in rat model of gastrointestinal motility disorder. Methods Seventy Wistar rats were randomly divided into groups of control, model, water extractive of PCR, hesperidin, water extractive of PCB, water extractive of PCB plus volatile oil and patchouli alcohol, ten rats in each group. Except the control group, the rest groups were established gastrointestinal motor disorder model via limb ischemia-reperfusion (LIR). After modeling rats of groups were intervened with corresponding extracts. The effects of different extracts on contraction amplitude of corpora ventriculi and small intestine smooth muscle were observed. The levels of gastrointestinal hormones including motilin (MOT), gastrin (GAS), cholecystokinin (CCK) and somatostatin (SS) were detected by radioimmunoassay. Results The contraction amplitudes of corpora ventriculi and small intestine smooth muscle were decreased (P<0.05), the serum level of GAS and plasma level of MOT were also significantly decreased, while CCK and SS levels in the gastric antrum were significantly increased in model group than those of the control group (P < 0.05). Water extractive of PCR, hesperidin, water extractive of PCB and water extractive of PCB +patchouli oil can increase the contraction amplitudes of corpora ventriculi and small intestine smooth muscle, increase the serum level of GAS and reduce levels of CCK and SS in the gastric antrum (P<0.05), whereas showed no influence in the plasma level of MOT (P>0.05)]. Compared with model group, patchouli alcohol showed no influence in the contraction of gastrointestinal smooth muscle and levels of MOT, GAS, CCK and SS (P>0.05). In the aspect of regulating the contraction of gastrointestinal smooth muscle and the level of GAS, CCK and SS, the pharmacological effect of PCR water extract was better than that of hesperidin (P<0.05), while water extractive of PCB+volatile oil was better than that of water extractive of PCB (P < 0.05). Conclusion The active ingredients of PCR and PCB have variant regulative effects on the contraction of gastrointestinal smooth muscle and the serum level of GAS, CCK and SS in the gastric antrum in rat model of gastrointestinal motility disorder.

OBJECTIVETo study Twist expression in thyroid papillary carcinoma (PTC) by immunohistochemistry and to assess its usefulness as marker in the differential diagnosis of PTC, follicular adenomas (FA) and benign papillary lesions (BPL).

METHODSFifty cases of PTC, 48 cases of FA and 47 cases of BPL were evaluated using manual tissue chip and SP immunohistochemical stain to detect the expression of Twist and HBME-1, and comparing the staining to that of cytokeratin 19 (CK19).

RESULTSIn PTC, positive rates of Twist, HBME-1 and CK19 were 100% (48/48), 94.0% (47/50) and 78.0% (39/ 50) respectively; in FA, positive rates were 0, 6.7% (3/45) and 0 respectively; in BPL, positive rates were 7.0% (3/34), 2.1% (1/47) and 0, respectively. The differences between PTC and FA and between PTC and BPL were both statistically significant (P = 0. 000). The sensitivity of Twist, HBME-1 and CK19 was 100%, 94.0% and 78.0%; the specifity was 96.4%, 95.7% and 100%; overall accurary was 97.7%, 95.1% and 91.9%, respectively.

CONCLUSIONSPositive rates of Twist is higher than the other markers in PTC. Immunohistochemical staining of Twist has important significance in the differential diagnosis of thyroid lesions. Twist immunohistochemistry maybe helpful in diagnosis and differential diagnosis of PTC.

Adenocarcinoma, Follicular ; metabolism ; Adenocarcinoma, Papillary ; pathology ; Adenoma ; diagnosis ; metabolism ; Biomarkers, Tumor ; immunology ; Carcinoma, Papillary ; diagnosis ; metabolism ; pathology ; Carcinoma, Papillary, Follicular ; metabolism ; Diagnosis, Differential ; Galectin 3 ; genetics ; metabolism ; Immunohistochemistry ; Keratin-19 ; genetics ; Keratins ; genetics ; metabolism ; Nuclear Proteins ; genetics ; metabolism ; Thyroid Neoplasms ; diagnosis ; metabolism ; pathology ; Thyroid Nodule ; pathology ; Twist-Related Protein 1 ; genetics ; metabolism