Objective To explore clinical application of single utility port or single port video-assisted thoracoscopic lobectomy.Methods Conducting a prospective study to 191 patients with lung disease who underwent complete video-assisted thoracoscopic surgery (VATS) lobectomy in Department of Thoracic Surgery of The People's Hospital of Wuhan University from June 2013 to Dec 2014.Results Of the 191 patients, 35 underwent left upper lobectomy, 42 underwent left lower lobectomy, 43 underwent right upper lobectomy, 11 underwent right middle lobectomy, 47 underwent right lower lobectomy, and 13 underwent bilobectomy.Operations were successful in all patients with 3 patients transferred for open thoracotomy for severe adhesion or bleeding.A total of 7 of 78 upper lobectomy and 8 of 113 middle or lower lobectomy was done with adding another incision for severe adhesion.The mean operative time was 50 ～ 190 (80.3 ±43.2) min.The mean blood losing was 90 ～ 350 (145.4 ± 56.2) ml.Thirty nine patients underwent upper lobectomy, who were placed two chest tubes, respectively.The upper chest drainage duration was (1.5 ±0.8) d, and the lower chest drainage duration was (4.2 ± 1.3) d.Forty eight patients underwent lower lobectomy, middle lobectomy, or bilobectomy, who were placed one chest tube, respectively.The chest drainage duration was(4.0 ± 1.7)d.The mean recovery time after operation was (6.1 ± 2.5) d.Seventy six patients were diagnosed with lung cancer, and the average number of dissected lymph nodes from each patient was (14.7 ±6.9).The lung cancer was classified as tumor node metastasis (TNM) stage Ⅰ A, Ⅰ B,ⅡA, ⅡB and ⅢA in 67, 61, 34, 22 and 7 cases, respectively.No serious complications, such as bronchopleural fistula or death, occurred in perioperation.Conclusions In consideration of placing two chest tubes after upper lobectomy and placing one chest tube after lower lobectomy, middle lobectomy, or middle and lower lobectomy, we think single utility port-VATS (2-port) for upper lobectomy and single port-VATS lobectomy for lower lobectomy, middle lobectomy, or middle and lower lobectomy are technically safe and have the advantages of drainage, lessening pain, rapid postoperative recovery, and have no significant difference in operation time, the incidence of complications and the number of removed lymph nodes, compared to traditional 3-port-VATS.

An HPLC method for the determination of 18alpha-glycyrrhetinic acid and 18beta-glycyrrhetinic acid in rat plasma was established, which was used subsequently to determine the pharmacokinetic profiles of both epimers of glycyrrhetinic acid in rats. alpha-glycyrrhetinic acid, beta-glycyrrhetinic acid, and a mixture of alpha-glycyrrhetinic and beta-glycyrrhetinic acids were administered to rats via gastric infusion. Blood samples were collected at different time intervals and extracted by liquid-liquid extraction. Separation was achieved by using a Kromasil C18 column (150 mm x 4.6 mm, 5 microm) with the mobile phase composed of acetonitrile--4 mmol x L(-1) ammonium acetate solution (46 : 54, v/v) at a flow rate of 1.0 mL x min(-1), and the detection wavelength was set at 250 nm. The pharmacokinetic parameters were calculated using the software DAS 2.0. In a combined administration, the main pharmacokinetic parameters of beta-glycyrrhetinic acid are significantly different from that of alpha-glycyrrhetinic acid (P < 0.05), while no significant difference was obtained when administrated individually. Compared to the single administration, significant differences (P < 0.05) on the values of AUC(0-t) and AUC(0-infinity) of beta-glycyrrhetinic acid were observed when this chemical was administrated together with alpha-glycyrrhetinic acid. In contrast, the pharmacokinetic parameters of alpha-glycyrrhetinic acid were not affected even under the co-administration. Here, a sensitive, specific, rapid and reproducible HPLC method was developed for the pharmacokinetic studies of alpha-glycyrrhetinic acid and beta-glycyrrhetinic acid in rat plasma.

Objective （ ） To explore the application value of bone suppression imaging BSI in the diagnosis of occupational （ pneumoconiosis） Methods - pneumoconiosis hereinafter referred to as " " . A total of 330 chest films of high kV digital （ ） radiograph DR of patients with suspected pneumoconiosis were selected by convenient sampling method. BSI is applied to the ， ， ， ， chest films and the differences of small opacity shape small opacity aggregation the number of large opacity lung areas small （ ）， opacity profusion and diagnostic stage of pneumoconiosis were analyzed by simple DR reading DR group simple BSI reading （ ） （ ） Results BSI group and DR and BSI combined reading combined group . There was no significant difference in the distribution of small shadows and the detection rate of small shadows aggregation and large shadows in pneumoconiosis among （ P ） ， the three film reading methods all >0.05 . For the concentration distribution of each lung area there was statistically （P< ）， significant difference between the DR group and the BSI group 0.05 but there was no statistically significant difference ， （ P ） between the DR group and the combined group and between the BSI group and the combined group all >0.05 . The results of ， consistency analysis showed that the DR group and the BSI group and the DR group and the combined group had high （ ， P< consistency in the judgment of small shadow intensity in the lung region both weighted Kappa coefficient were 0.75 all ） 0.01 . There was a high consistency between BSI group and DR group and combined group and DR group in the diagnosis of （ ， ， P< ） ， pneumoconiosis stage weighted Kappa coefficient were 0.77 0.79 all 0.01 . Compared with the DR group the diagnostic ， rate of pneumoconiosis stage Ⅰwas significantly reduced and the diagnostic rate of pneumoconiosis stage Ⅱ was significantly （ P< ） ， increased in the BSI group and the combined group all 0.01 . However there was no significant difference in the diagnosticrate of pneumoconiosis stage Ⅲ >0.05 . Both the BSI reading and DR and BSI combined reading can improve ， the display of pneumoconiosis lesions to varying degrees and therefore can improve the diagnosis of pneumoconiosis. In ， addition the identification and diagnosis of pneumoconiosis lesions in the BSI reading is comparable to that in the combined ， group which has a good application value in the diagnosis of pneumoconiosis.

Objective To explore the effects of coriafia lactone (CL)-activated astrocytes (Ast) conditioned medium (ACM) on the expressions of glutamate (Glu) and GluR2 in the brain of rat. Methods Asts of hippocampus were cultured according to the McCarthy and DeVellis's method, and then the ACM was collected. Forty-eight male adult Sprague-Dawley (SD) rats were randomly divided into the control group (n=16) and the CL group (n=32). Rats in the control group were administered 10 μL ACM I. C. V., which was not added any stimulating substance. Rats of the CL group were injected I. C. V. 10 μL CL-activated ACM. The rats in both groups were subdivided into post-injection 2,4,8,12h subgroups, 4 in each subgroup in the control group and 8 in each subgroup in the CL group. The behaviors of the rats were observed and the expressions of Glu and GluR2 in the cerebral cortex and hippocampus were detected with immunohistochemistry and immunofluorescence. The content of GluR2 was tested with Western blot. Results The rats injected with CL-activated ACM showed seizure activities, whereas the rats of the control group showed no seizure activities. The expression of Glu in cerebral cortex and hippocampus in the brains injected with CL-activated ACM was increased compared with the control group at 4h (P<0.05), but the expression of GluR2 was attenuated compared with the control group at 4h(P<0.05). The results of GluR2 in the cerebral cortex and hippocampus detected with Western blot were different significantly with control group (P<0.05). Conclusion CL-activated ACM can enhance the expression of Glu and reduce the expression of GluR2 in the brain of rat, resulting in the activation of AMPA pathway and the Ca2+ influx, and then induce seizure activities.

Animals ; Biphenyl Compounds ; pharmacology ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Genetic Therapy ; Hepatocyte Growth Factor ; pharmacology ; Hepatocytes ; drug effects ; Humans ; Liver Cirrhosis ; pathology ; therapy ; Plants, Medicinal ; chemistry ; Ursodeoxycholic Acid ; pharmacology

This study reported the obtainment of the full-length cDNA of Salvia miltiorrhiza hairy roots (Abbr: SmHDR, GenBank number: JX233817), via extracting Salvia miltiorrhiza hairy roots total RNA, designing specific primers according to the transcriptome data and using the RACE strategy, and then analyzed it with bioinformatics approaches. On this basis, using the real-time PCR to detect SmHDR gene expression after Ag+ induction, and testing tanshinones contents of corresponding samples by UPLC. SmHDR has 1 647 nucleotides, and an open reading frame (ORF) encoding a protein of 463 amino acid residues. The deduced protein has isoelectric point (pI) of 5.72 and a calculated molecular weight about 51.88 kD. In the secondary structure, the percentage of alpha helix, beta turn and random coil were 35.64%, 20.30% and 44.06%, respectively. Sequence alignment and phylogenetic analysis demonstrated that SmHDR had relative close relationship to the HDR of Picrorhiza kurrooa, similar to HDR from other species of plants. Real time PCR results indicated that elicitor of Ag+ stimulated the increase of mRNA expression of SmHDR. At the same time, results of ultra performance liquid chromatography (UPLC), used to examine the accumulation of diterpenoid tanshinones in hairy roots, showed that the contents of diterpenoid tanshinones in hairy roots of Salvia miltiorrhiza were increased dramatically at 12 h after treated with Ag+, and then decreased significantly. This result showed a positive correlation between the levels of mRNA expression and tanshinones accumulation in Salvia miltiorrhiza stimulated by Ag+. The content of tanshinones was gradually raised, and it had an obvious increase at 120 h. The bioinformatics analysis and gene expression indicated that SmHDR might be involved in tanshinones biosynthesis, which laid the foundation for further study of secondary metabolic regulation mechanism of tanshinones.
Amino Acid Sequence ; Cloning, Molecular ; DNA, Complementary ; genetics ; Diterpenes, Abietane ; biosynthesis ; metabolism ; Gene Expression Regulation, Plant ; Open Reading Frames ; Oxidoreductases ; chemistry ; genetics ; metabolism ; Phylogeny ; Plant Roots ; genetics ; metabolism ; Plants, Medicinal ; genetics ; metabolism ; Protein Structure, Secondary ; RNA, Messenger ; metabolism ; Salvia miltiorrhiza ; genetics ; metabolism ; Sequence Alignment ; Silver Nitrate ; pharmacology ; Synthetic Biology

OBJECTIVETo determine the effectiveness of surgical correction for post-traumatic kyphosis of thoracolumbar spine.

METHODSFrom 1996 to 2003, 33 consecutive patients with post-traumatic kyphosis of thoracolumbar spine were corrected surgically. The mean age was 40.3 years (range, 13 - 65 years). The mean time between the initial injury and surgical correction was 36.0 months (range, 6 - 220 months). The kyphotic deformity averaged 40.8 degrees (range, 20 degrees - 82 degrees ). All the patients had neurological deficits. Twelve patients had obvious back pain. Seven patients lost sphincter function completely and nineteen patients lost the function partly. Twenty-three patients had ever undergone laminectomy and/or instrumentation. The treatment procedure consisted of anterior release and posterior spinal osteotomy with instrumentation (15 patients), posterior closing wedge osteotomy with instrumentation (12 patients), anterior release and instrumentation (6 patients).

RESULTSKyphosis was corrected from an average of 40.8 degrees to an average of 5.7 degrees, the corrective rate was 86.0% (40.8 degrees - 5.7 degrees /40.8 degrees). There were no severe complications. The average follow-up period was 24.6 months (range, 6 - 84 months). There was no loss of correction at follow-up. Ten of these patients showed an improvement in neural function by one or two levels according to the classification. Sphincter function recovered partly in ten patients. Back pain was relieved significantly in all of twelve patients with back pain preoperatively. Bony fusion was achieved in thirty-two patients. One patient had nonunion and achieved bony fusion after revision.

CONCLUSIONPosterior closing wedge osteotomy was suitable to kyphosis less than 40 degrees. Anterior release and posterior spinal osteotomy was effective, especially to the patients with severe kyphosis deformity or with operation history. Patients with incomplete neurological deficits and/or severe back pain could get benefit from osteotomy of spine, even if their medical history was long.

Adolescent ; Bone Transplantation ; Child ; Female ; Humans ; Kyphosis ; etiology ; surgery ; Lumbar Vertebrae ; injuries ; surgery ; Male ; Middle Aged ; Osteotomy ; methods ; Retrospective Studies ; Spinal Fractures ; complications ; surgery ; Spinal Fusion ; Thoracic Vertebrae ; injuries ; surgery ; Treatment Outcome

OBJECTIVETo evaluate the efficacy of allogenic strut bone graft and instrumentation for anterior cervical fusion following subtotal corpectomy and decompression in cervical myelopathy.

METHODSThirty-five patients with cervical myelopathy were treated by the procedure of allogenic strut bone graft and instrumentation for anterior cervical fusion following subtotal corpectomy and decompression. The preoperative average JOA scale score was 8.7 point (Range 4-15).

RESULTSSixty-nine vertebral were corpectomized and 104 levels were decompressed and fused with an average of 3 levels. Among the cases, 1 vertebrae was corpectomized in 7 cases, 2 vertebra in 22 cases, 3 vertebra in 6 cases. There were no surgery-related complications. The patients were followed up from 11-37 months, with an average of 17.4 months. No plate breakage, screw loose, graft infection, lysis and absorption was discovered. The fusion rate was 100%, the average time of fusion was 9.3 months (range from 6-15 months). The postoperative average JOA scale score was 14.8 point (range 7-17), the recovery ratio was 73.5% and the excellent and good results was 82.8%.

CONCLUSIONSThe use of allogenic strut bone graft and instrumentation for anterior cervical fusion following subtotal corpectomy and decompression in cervical myelopathy may not only simplify surgical procedure and decrease injuries and complications, but also the fusion is satisfactory and reliable.

Bone Transplantation ; Cervical Vertebrae ; surgery ; Female ; Follow-Up Studies ; Humans ; Laminectomy ; Male ; Spinal Fusion ; Spinal Osteophytosis ; surgery ; Transplantation, Homologous ; Treatment Outcome

Objective: To explore the effect of impaired cardiac microvascular endothelial cells on rat's embryo cardiomyocyte apoptosis and the impact of Tongxinluo in such process. Methods: 3 kinds conditioned medium of cardiac microvascular endothelial cells were prepared. ① normal rat's cardiac microvascular endothelial cell (RCMECs) conditioned medium, ②homocysteine induced impaired RCMECs conditioned medium and ③ Tongxinluo treated RCMECs conditioned medium. Rat's embryo H9c2 cardiomyocytes were divided into 4 groups:① Control group, cells were cultured in DMEM without serum, ② N-CM group, cells were cultured in RCMECs conditioned medium, ③ H-CM group, cells were cultured in homocysteine induced impaired RCMECs conditioned medium and ④ T-CM group, cells were cultured in Tongxinluo treated RCMECs conditioned medium. Cell mitochondrial membrane potential (MMP) was detected by JC-1 agent, ROS level was examined by flow cytometry, cardiomyocytes apoptosis was measured by live cell imaging system, expression levels of cytochrome C (Cyt-C), cysteinyl aspartate specific proteinase-3 (Caspase-3), Bcl-2 associated X protein (Bax) and B-cell leukemia/lymphoma 2 protein (Bcl-2) were detected by Western blot analysis. Results: MMP, expression levels of ROS, Cyt-C and cardiomyocyte apoptosis were similar between Control group and N-CM group, P>0.05. Compared with N-CM group, H-CM group had decreased MMP, increased ROS and cardiomyocyte apoptosis, up-regulated expressions of Cyt-C, Caspase-3 and Bax, all P<0.01. Compared with H-CM group, T-CM group had increased MMP, decreased ROS and cardiomyocyte apoptosis, down regulated expressions of Cyt-C, Caspase-3 and Bax, all P<0.05; while up-regulated expression of Bcl-2, P<0.01. Conclusion: Conditioned medium of impaired cardiac microvascular endothelial cells may induce cardiomyocyte apoptosis and such process could be inhibited by Tongxinluo.

Objective：To investigate the correct expression of dengue 2 virus 43 strain NS1 gene in transfected BHK-21 cell. Methods：The D2-43 DNA fragment coding for signal peptide plus NS1 protein was cloned between KpnⅠ site and EcoR Ⅰ site of expression plamid pcDNA3.1. The obtained recombinant vector pcDNA-NS1 was transfected into BHK-21 cells with electroporation technique. After selection by G418, resistant clones were screened by RT-PCR and Western blotting test. Results：The RT-PCR results of four in five randomly selected cell clones were positive. Western blotting test showed that NS1 gene could be expressed in BHK-21 cells. Conclusions：NS1 protein was capable of being expressed and appropriately processed in pcDNA-NS1 transfected BHK-21 cells. The present results suggest the feasibility of NS1-based DNA immunization.