Objective To evaluate improved effect for deep venous valve function after superficial vein surgery of lower extremity in the intermediate stage.Methods Totally 43 patients (55 limbs) with varicose veins of lower extremity were enrolled to accept surgical management of vein systems in our department from March 2006 to October 2006.All patients were respectively followed up after 6 months and 4 years about the changes of deep venous valve function with color Doppler ultrasonography.Results Thirty-nine patients’ deep venous valve function kept well up to now,and there was no significant difference between the two results.Four patients without proximal saphenous vein ligation recurred,and there was reflux in deep venous.Conclusion Endovenous laser treatment and ablation of varicose veins of lower extremity with deep venous insufficiency could improve deep venous valve function effectively.Proximal great saphenous vein ligation is important for successful operation.

Objective To evaluate thoracic endovascular aortic repair for type B aortic dissection.Methods Chnical data were reviewed on 126 cases with type B aortic dissection undergoing endovascular aortic repair in our hospital from January 2006 to April 2011.There were 86 male patients and 40 female patients,age from 32 to 82 years.The stent- grafts were introduced via femoral arteriotomy.Stent-grafts were implanted to blockade tears of aortic dissection under the guidance of DSA.Postoperatively patients were followed-up by angiography and imiging for endoleak,stent migration,and fracture of stent-graft.Results In all cases,the aortic dissection tears were closed,true lumens were opened,and organ function was restored.There were not complications such as internal hemorrhage due to trauma or stent displacement on CTA from 3 to 63 months after endovascular therapy.Procedure was successful in all 126 cases,157 stents were released,2 cases died in the perioperative period,1 case died during the followed-up.A breach was found at the end of the stent in 12 cases,endovascular aortic repair redone successfully with a retrograde type A dissection found during follow-up.Conclusions Endovascular aortic repair is safe and effective for patients with Stanford type B aortic dissection with a favorable outcomes.

Objective To establish the paclitaxel-resistant gastric cancer cell(HGC-27/PTX) and to investigate the changes of characteristics before and after resistance,as well as the possible resistant mechanisms.Methods The paclitaxel-resistant gastric cancer cell HGC-27/PTX was established by increasing paclitaxel dose gradually and intermittently.The IC50 (50% inhibitory concentration) and cell cycle were determined by CCK-8 assay and flow cytometry,respectively.The differentially expressed genes (DEGs) and signaling pathways were analyzed using RNAseq.Results The establishment of HGC-27/PTX cells lasted 9 months,and the sensitivity of paclitaxel of HGC-27/PTX cells was significantly lower than parental cells (P<0.05).Compared to parental cells,the morphology of HGC-27/PTX cells was slightly different,and the proportion of S and G2/M phase was obviously increased (P<0.01).A total of 274 DEGs were identified between the resistant and parental cells with 130 genes up-regulated and 144 genes down-regulated.DEGs were significantly enriched in extracellular matrix (ECM)-receptor interaction(P<0.001) and PI3K-Akt signaling pathways (P<0.05),which could provide evidences for reversing paclitaxel resistance.Conclusions The paclitaxel-resistant gastric cancer cells HGC-27/PTX was established with stable culturein vitro,which provides an ideal model for future study on the mechanism of drug resistance.

Objective:To investigate the perfection and improvement of the execution of integrative medicine therapy in severe tetanus therapy, to successfully control tetanus severe spasms, autonomic dysfunction and prevent lethal side-effect of prolong and high-dosage sedative-muscle-relaxant therapy, resulted in significant reduction of mortality of tetanus.Methods:Symptoms, treatments and outcome of tetanus patients admitted to Peking University Third Hospital from 1965 to 2020 were reviewed. Patients were classified with Ablett classification. The cases of Ablett grade Ⅲ and Ⅳ were severe tetanus. The patients were divided into two groups according to whether they were treated together with traditional Chinese medicine (TCM) simultaneously during the standard tetanus treatment; the patients in the TCM group were divided into the tetanus TCM medication group and the non tetanus TCM medication group according to the medicine provided whether was in accord with the conventional tetanus TCM prescriptions. The mortality of each group was calculated. In addition, one survived and one deceased case with severe convulsion, autonomic nerve dysfunction (Ablett grade Ⅳ) were selected, combined with the treatment methods and curative effects, the types, use methods and outcomes of Chinese and Western medicine were analyzed.Results:The 46 tetanus cases were treated with Western medicine. Twenty-two of them, TCM were applied. Fifteen of the 22 cases took the TCM prescription which was accord with the conventional tetanus prescription. The mortality of the 46 cases was 21.7% (10/46). The number of non-TCM group was 24 cases, with mortality of 20.8% (5/24); 1 case was Ablett Ⅱ, 1 was Ablett Ⅲ and 3 were Ablett Ⅳ. The number of the TCM group was 22 cases, with mortality of 22.7% (5/22), 2 cases were Ablett Ⅲ, 3 were Ablett Ⅳ. The TCM prescription of these 5 deceased cases was not directed towards tetanus. The tetanus TCM medication group was 15 cases, with no mortality. Case analyses: case 1 was intubated because of severe spasms. Autonomic dysfunction occurred on the 8th day after admission. Esmolol with increasing the dosage of the sedatives and muscle relaxant, was not effective. Tetanus TCM was applied after 2 days of autonomic dysfunction happened. Autonomic dysfunction was then under controlled on the 2nd day post-TCM. She was recovery and discharged after 4 weeks. Case 2, also was intubated because of severe spasms. Autonomic dysfunction happened on the 3rd day after admission, and failed to be controlled by large-dose sedatives, muscle relaxant, and Esmolol. After 8 days of persistent autonomic dysfunction, tetanus TCM was applied and autonomic dysfunction was under controlled on the 2nd day post-TCM administration. Large dosage of muscle-relaxant was applied continuously. After 5 days' administration of TCM, the TCM was withdrew. One day after the withdrawal of TCM, respiratory and cardiac arrest happened because of the diffused bronchiole obstruction with pulmonary secretions loading.Conclusion:Based on the precise and real-time diagnosis of the state of the disease, integrative medicine therapy with an overall analysis tetanus TCM prescription, is the key of declining tetanus mortality.

Amino Acids ; blood ; Blood Proteins ; analysis ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Infant, Newborn ; Intestinal Obstruction ; surgery ; Intestinal Perforation ; surgery ; Male ; Parenteral Nutrition ; Postoperative Care

OBJECTIVETo investigate the genetic polymorphism of human cytomegalovirus (HCMV) glycoprotein H (gH) from infantile clinical isolates, to analyze the genotypic distribution of gH in different diseases of HCMV infection and try to find the correlations between the diseases and genotypes.

METHODFresh urine specimens were collected from the hospitalized children with different diseases whose blood HCMV-IgM and HCMV-IgG were positive. Virus was isolated from these specimens. Glycoprotein H of harvest clinical isolates was genotyped by nested-PCR combined with restriction fragment length polymorphism (RFLP), the purified PCR products were digested by restriction endonuclease HhaI. The digested products were genotyped by polyacrylamide gel electrophoresis and silver staining. Classification and results of sequencing were compared.

RESULTTotally 102 HCMV clinical isolates were obtained. Glycoprotein H gene of these clinical isolates (43 cases had infantile hepatitis syndrome, 38 cases had anicteric hepatitis, 13 pneumonia, 7 thrombocytopenic purpura, and 1 congenital CMV infection) were positive by nested-PCR, whose positive rate was 100%. The results showed that 62 strains were gH1 genotypes (60.8%), while 40 strains were gH2 (39.2%), mixed type or new genotype was not observed. In infantile hepatitis syndrome (26 clinical isolates were gH1 genotypes, 17 clinical isolates were gH2 genotypes), anicteric hepatitis (25 were gH1, 13 were gH2) and pneumonia (9 were gH1, 4 were gH2), the distribution of HCMV gH genotypes of infantile clinical isolates was consistent with the overall trend (χ(2) = 0.357, P > 0.05). However , the gH2 was more common than gH1 in the clinical isolates of patients with thrombocytopenic purpura (6 were gH2, 1 were gH2, χ(2) = 6.083, P < 0.05).

CONCLUSIONGenotype 1 was the dominant genotype of glycoprotein H in HCMV clinical isolates from our hospital infants. There was no significant difference between the distribution of gH genotypes in infantile hepatitis syndrome, anicteric hepatitis and pneumonia. However, gH2 was the dominant genotype in thrombocytopenic purpura. These findings suggested that there may be a certain relevance between gH genotype and different clinical manifestations.

Amino Acid Sequence ; Base Sequence ; Child, Preschool ; Cytomegalovirus ; classification ; genetics ; isolation & purification ; Cytomegalovirus Infections ; virology ; DNA Primers ; DNA, Viral ; genetics ; Female ; Genotype ; Hepatitis ; virology ; Humans ; Infant ; Infant, Newborn ; Male ; Pneumonia, Viral ; virology ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length ; Urine ; virology ; Viral Envelope Proteins ; genetics

Objective To investigate the effect of nicotine on proliferation and chemosensitivity of A549 cells in vitro. Methods A549 cells was assessed by MTT assay to measure cell proliferation and was assessed by RT-PCR tomeasure chemosensitivity. Results 0.01~100μmol/L nicotine could promote the proliferation of A549 cells, the most marked proliferation at 1μmol/L, compared with the control group, the activity of A549 cells was increased by 1.85 times (P<0.01). When the concentration of nicotine above 1μmol/L, the proliferation of A549 cells had an decreasing tendency. When the concentration above 1000μmol/L, the proliferation of A549 cells can be inhibited. Nicotine can also reduce chemosensitivity of A549 cells to 5-FU, with the addition of nicotine, A549 cells survival rate increased significantly, the most marked at 1μmol/L, compared with the control group, the inhibitory rate of A549 cells was 9 % (P< 0.01). Nicotine significantly increased the expression level of α7 nAChR in A549 cells and decreased the expression of PTEN , in a concentration dependent manner. Compared with the control group, 1μmol/L of nicotine could increase the expression levels of α7 nAChR by 3.4 fold, and decrease the expression levels of PTEN by 60.36 % (P< 0.01). Conclusion Nicotinecan promote the growth of A549 cells and reduce chemosensitivity of A549 cells to 5-FU.

Lysine decarboxylase is a key enzyme involved in the upstream biosynthesis of lycopodium alkaloids (LAs) such as huperzine A, contributing to the decarboxylation of lysine to 1,5-pentanediamine (cadaverine). Three lysine decarboxylase genes (HsLDC-L1, HsLDC-L2, HsLDC-L3) were successfully cloned from Huperzia serrata using transcriptomic sequence data mining strategy combined with reverse transcription PCR. The physicochemical properties, secondary and tertiary structures, amino acid identities, and evolutionary relationship of the three LDCs were analyzed by online bioinformatics analysis platforms and DNAMAN, MEGA 7.0 software, revealing that all of these proteins had the conserved PLP binding domain and active site residues were completely conserved in LDCs. Phylogenetic analysis showed that these LDCs were located in the same branch as other known LDCs from LA-producing plants. Accordingly, the ORFs of these three HsLDCs were inserted into different expression plasmids for further expression in E. coli. However, only HsLDC-L1 was successfully expressed in E. coli BL21 (DE3) by inserting into a pCold TF vector. The recombinant protein was purified by Ni²⁺ affinity chromatography purification. HsLDC-L1 contains 469 amino acid residues, with a calculated molecular weight of 50.50 kDa. HsLDC-L1 expectedly catalyzed the decarboxylation of lysine to produce cadaverine. In addition, HsLDC-L1 can also catalyze the generation of putrescine from ornithine. However, it cannot catalyze the decarboxylation of tyrosine, phenylalanine, tryptophan and histidine. The results not only provide insight into the biosynthesis of LAs including huperzine A, but also provide a critical genetic element for the overproduction of Δ¹-piperideine and pelletierine, the essential biosynthetic precursors of LAs, using synthetic biology strategies.

BACKGROUNDThe production of neural stem cells (NSCs) derived from embryonic stem (ES) cells was usually very low according to previous studies, which was a major obstacle for meeting the needs of clinical application. This study aimed at investigating whether astrocytes could promote production of NSCs derived from ES cells in vitro.

METHODSMouse ES cells line-D3 was used to differentiate into NSCs with astrocytes as inducing stromal cells by means of three-stage differentiation procedure. Another group without astrocytes served as control. The totipotency of ES cells was identified by observation of cells' morphology and formation of teratoma in severe combined immunodeficiency disease (SCID) mice. The quantity and purity of NSCs derived from ES cells were analyzed using clonogenic assay, immunohistochemical staining and flow cytometry assay. The plasticity of NSCs was detected by differentiating test. Octamer-binding transcription factor 4 (Oct-4) and nestin, the specific marker genes of ES cells and NSCs respectively, were detected continuously using reverse transcription-polymerase chain reaction (RT-PCR) method to monitor the process of cell differentiation.

RESULTSThe ES cells of D3 line could maintain the ability of differentiating into cellular derivations of all three primary germ layers after continuous passage culture. At the end of two-stage of inducing process, 23.2 +/- 3.5 neurospheres per plate formed in astrocyte-induced group and only 0.8 +/- 0.3 per plate in the control group (clonogenic assay, P < 0.01), and the ratio of nestin positive cells was (50.2 +/- 2.8)% in astrocyte-induced group and only (1.4 +/- 0.5)% in the control group (flow cytometry, P < 0.01). With the induction undergoing, the expression of Oct-4 gradually decreased and then disappeared, while the expression of nestin was increased step by step, and the ratio of nestin positive cells was up to 91.4% by the three-stage differentiation. The nestin positive cells could be further induced into neurons, astrocytes, and oligodendrocytes in differentiating medium supplemented with fetal calf serum. The results of differentiating test showed that the ratio of NF-200 and NSE positive cells was (42.7 +/- 2.6)% in astrocyte-induced group and only (11.2 +/- 1.8)% in the control group (P < 0.01).

CONCLUSIONSAstrocytes can not only increase the production of NSCs derived from ES cells but also promote the differentiation of NSCs toward neuronal lineage.

Animals ; Astrocytes ; physiology ; Cell Differentiation ; Cell Lineage ; Cells, Cultured ; Embryo, Mammalian ; cytology ; Mice ; Neurons ; cytology ; Stem Cells ; cytology

Objective To discuss the application of double parallel stent for preserving arch branch vessels during thoracic endovascular aortic repair.Methods During the period from June 2015 to June 2016,double parallel stents were carried out in 9 cases at department of vascular surgery,the People's Hospital of Xinjiang Uygur Autonomous Region.Results One patient died of an acute ischemic stroke and subsequently resulted in multiple organ failure after thoracic endovascular aortic repair.Eight patients received an aorta CT angiography during follow up at 1,3,6 months.During follow up,the patency rate for double parallel stents was 100％.There were no endoleak in 8 cases.Conclusion Double parallel stents are suited to patients with emergency situation and unsuited to open surgery in complicated aortic arch pathologies.Surgical approach is determined by preoperative evaluation.Prevention of endoleak and stroke is important in all cases.