Acupuncture Therapy ; Adolescent ; Adult ; Aged ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Male ; Middle Aged ; Myasthenia Gravis ; drug therapy ; therapy ; Young Adult

Objective To investigate the clinical efficacy of laparoscopic hepatectomy.Methods The clinical data of 258 patients who received laparoscopic hepatectomy at the First Affiliated Hospital of Fujian Medical University from March 2010 to January 2013 were retrospectively analyzed.There were 196 patients with primary hepatic cancer,45 with hepatic hemangioma,13 with hepatic focal nodular hyperplasia,2 with hepatic metastatic cancer,1 with carcinoma of gallbladder and 1 with hepatic hamartoma.All patients were followed up via phone call or out-patient examination till March 2013.Results A total of 142 patients received single hepatic segmentectomy,98 received multiple hepatic segmentectomy,18 received multiple lesions resection.Fifty-one patients received hepatic tumorectomy + cholecystectomy.All the operations were successfully done under laparoscope without conversion to the open surgery.The mean tumor diameter and the operation time were (5 ± 3) cm (range,1.0-11.5 cm) and (113 ± 56) minutes (range,50-310 minutes),respectively.Intraoperative hepatic portal occlusion was performed on 122 patients,and the time for hepatic portal occlusion was (15 ± 7)minutes.The volume of intraoperative blood loss was (211 ± 195)mL (range,10-650 mL),and blood transfusion was not needed.The capsule of the tumor was complete.The distance between the resection margin and the malignant tumor was above 1.5 cm,and there was no residual tumor in the resection margin.The hepatic function was back to the normal level in 1 week after the operation,and no patient had hepatic failure.The duration of postoperative hospital stay was (7.2 ± 1.3)days (range,5-10 days).One patient was complicated with bile leakage,6 with slight peritoneal effusion,and other patients had no postoperative complications.The rate of follow-up was 91.47％ (236/258),and the time of follow-up was (16 ± 10) months.A total of 199 patients with malignant hepatic tumors were followed up.During the follow-up,180 patients had tumor-free survival; 18 patients had postoperative tumor recurrence; 1 patient had omental metastasis and received surgical resection.Thirty-seven patients with benign hepatic tumor survived without complication during the follow-up.Conclusion Laparoscopic hepatectomy is effective for the treatment of hepatic tumors.Multiple hepatic inflow occlusion under laparoscope in a short time may improve the safety of surgery,without prolonging the recovery time of patients.

Adolescent ; Adult ; Anticoagulants ; poisoning ; Coagulation Protein Disorders ; chemically induced ; diagnosis ; therapy ; Female ; Humans ; Male ; Rodenticides ; poisoning

The culture medium and cultural conditions of solid-state fermentation of Streptomyces Menmyco-93-63 were tested in this study. The suitable medium which contains rice, sorghum, millet bran, and rice hull with the proportion of 2:2:3:3 was developed for the spore production of Streptomyces Men-myco-93-63 using single substrate screening, mixture substrate screening and orthogonal experiments, and the sporulation was up to 2.52?109 CFU/g. And then, initial charge, initial ratio of water to solid, inoculating quantity, and culture temperature impact to sporulation of Streptomyces Men-myco-93-63 were tested. The favorite cultural conditions are developed as the following: the initial charge is 15 g in 500 mL Erlenmeyer flask; initial ratio of water to solid is 1.7:1.0 (V/W, rice hull excluding), inoculating quantity is 7 mL, culture temperature is 28℃.

Aim:To investigate the effect of Hemangiopoietin (HAPO) on the hematopoiesis reconstitution in sub-lethally irradiated Balb/c mice.Methods: Balb/c mice were underwent total body irradiation at 700 cGy 137Cs ? radiation and were treated with HAPO or recombinant human granulocyte colony stimulating factor (rhG-CSF) after irradiation. The hematopoiesis reconstitution of mice were detected. Cells from bone marrow of Balb/c mice were cultured with HAPO or rhG-CSF for 24 hours or 72 hours before or after the cells were irradiated. The viability of cells were assessed and the ability of in vitro hematopoiesis reconstitution were also detected. Result: rhG-CSF and HAPO treated mice both showed increased survival rate and increased colony forming units. The peripheral WBC number increased greatly. The HAPO group was most quickest compared with rhG-CSF group and PBS control group. The number of bone marrow cells at day 14 of rhG-CSF group was higher than that in HAPO group, but the number of bone marrow cells at day 32 of rhG-CSF was lower than that in HAPO group. The number of bone marrow cells at day 42 of rhG-CSF was below normal. The number of bone marrow cells at day 42 of HAPO group was nearly normal. The number of CFU-GEMM in HAPO group was most compared with that in rhG-CSF group and PBS control group at day 7, 14 and 21 after radiation. The survival rate of cells after radiation in HAPO group was markedly higher than that in PBS control group, but the survival rate of cells after radiation in rhG-CSF group was no notable difference compared with that in PBS control group. In MTT assay, both HAPO and rhG-CSF incubation stimulated proliferation of bone marrow cell at 72 hours after radiation. Bone marrow cells formed Hematopoietic islands in HAPO group after radiation and were positive for sca-1 and CD31. CD31 positive endothelial cells increased around the Hematopoietic islands. There was no Hematopoietic islands formation, few CD31 positive endothelial cells and no sca-1 positive cells in PBS control group. Conclusion: HAPO can promote hematopoiesis reconstitution in sub-lethally irradiated Balb/c mice. It can increase the survival rate of mice and stimulate the proliferation of hematopoietic stem cells.

Adult ; Case-Control Studies ; Female ; Humans ; Male ; Middle Aged ; Orthopedic Procedures ; Popliteal Cyst ; surgery ; Suture Techniques

OBJECTIVETo study the expression of Smo protein and the downstream transcription factor Gli1 protein in Sonic hedgehog signal transduction pathway in gastric carcinoma.

METHODSA tissue microarray was constructed using 85 gastric carcinoma and 25 normal gastric mucosa specimens. The expression of Smo and Gli1 proteins were detected immunohistochemically and the correlation between their expression in gastric carcinoma was analyzed.

RESULTSOnly weak expression, if any, of Smo and Gli1 proteins was detected in normal gastric mucosa, but in papillary adenocarcinoma, tubular adenocarcinoma and poorly differentiated adenocarcinoma, their expressions were significant increased as the differentiation degree was lowered. Smo protein expression in gastric carcinoma was significantly correlated with that of Gli1 protein with correlation coefficient of 0.989 (P<0.001).

CONCLUSIONThe abnormal activity of Sonic hedgehog signal transduction pathway may play an important role in the occurrence of papillary adenocarcinoma, tubular adenocarcinoma and poorly differentiated adenocarcinoma, and this abnormality is associated with Smo protein overexpression, which upregulates the expression of the downstream transcription factor Gli1 protein.

Adenocarcinoma ; metabolism ; pathology ; physiopathology ; Adult ; Aged ; Female ; Hedgehog Proteins ; physiology ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Receptors, G-Protein-Coupled ; biosynthesis ; Signal Transduction ; Smoothened Receptor ; Stomach Neoplasms ; metabolism ; pathology ; physiopathology ; Transcription Factors ; biosynthesis ; Zinc Finger Protein GLI1

OBJECTIVETo explore the possibility of repairing chemically induced acute hepatic injuries with allogeneic bone marrow stem cell (BMSC) transplantation.

METHODSA SD rat model of CCl(4)-induced acute hepatic injury was established, which received transplantation of BMSCs (2.0 ml, 1x10(6)/ml) or normal saline injection into the local liver parenchyma, respectively. The rats were sacrificed at 6 h before and 6 h, 1, and 5 weeks after transplantation, and the livers were prepared for microscopic examination.

RESULTSCellular necrosis, bridging necrosis, congestion in the hepatic sinusoid, and inflammatory cell infiltration were seen in the chemically injured livers 6 h after model establishment, and these changes were ameliorated in rats receiving BMSC transplantation.

CONCLUSIONSAllogeneic BMSC transplantation can repair chemically induced acute liver injuries.

Animals ; Bone Marrow Cells ; cytology ; Carbon Tetrachloride ; toxicity ; Chemical and Drug Induced Liver Injury ; etiology ; pathology ; surgery ; Hematopoietic Stem Cell Transplantation ; methods ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Treatment Outcome

The mutation of cystic fibrosis transmembrane conductance regulator (CFTR) gene leads to an autosomal recessive genetic disorder cystic fibrosis (CF). The gene therapy for CF using adeno-associated virus (AAV) vectors delivering CFTR gene is restricted by the contents limitation of AAV vectors. In this study the split CFTR genes severed at its regulatory domain were delivered by a dual-vector system with an intein-mediated protein trans-splicing as a technique to investigate the post-translational ligation of CFTR half proteins and its function as a chloride ion channel. A pair of eukaryotic expression vectors was constructed by breaking the human CFTR cDNA before Ser712 codon and fusing with Ssp DnaB intein coding sequences. After co-transfection into baby hamster kidney (BHK) cells followed by transient expression, patch clamps were carried out to record the chloride current of whole-cell and the activity of a single channel, and the ligation of two halves of CFTR was observed by Western blotting. The results showed that the intein-fused half genes co-transfected cells displayed a high whole cell chloride current and activity of a single channel indicating the functional recovery of chloride channel, and an intact CFTR protein band was figured out by CFTR-specific antibodies indicating that intein can efficiently ligate the separately expressed half CFTR proteins. The data demonstrated that protein splicing strategy could be used as a strategy in delivering CFTR gene by two vectors, encouraging our ongoing research program on dual AAV vector system based gene transfer in gene therapy for cystic fibrosis.
Animals ; Cells, Cultured ; Chlorides ; metabolism ; Codon ; genetics ; Cricetinae ; Cystic Fibrosis ; therapy ; Cystic Fibrosis Transmembrane Conductance Regulator ; genetics ; metabolism ; DNA, Complementary ; genetics ; Dependovirus ; genetics ; Genetic Therapy ; Genetic Vectors ; Humans ; Inteins ; physiology ; Kidney ; cytology ; Protein Processing, Post-Translational ; Recombinant Proteins ; genetics ; metabolism ; Trans-Splicing ; Transfection

We recently demonstrated that an intein-mediated protein splicing can be used to transfer B-domain-deleted FVIII (BDD-FVIII) gene by a dual-vector. In this study, we observed the effect of a variant heavy chain with six potential glycosylation sites of B domain and L303E/F309S mutations in its A1 domain, which were proven to be beneficial for FVIII secretion, on secretion of spliced BDD-FVIII. By transient co-transfection of cultured 293 cells with intein-fused variant heavy chain (DMN6HCIntN) and light chain (IntCLC) genes, the culture supernatant was analyzed quantitatively by ELISA for secreted spliced BDD-FVIII antigen and by a chromogenic assay for bioactivity. The data showed that the amount of spliced BDD-FVIII protein and coagulation activity in culture supernatant from DMN6HCIntN plus IntCLC co-transfected cells were up to (149 +/- 23) ng x mL(-1) and (1.12 +/- 0.14) u x mL(-1) respectively greater than that of intein-fused wild type heavy (HCIntN) and light chain (IntCLC) co-transfected cells [(99 +/- 14) ng x mL(-1) and (0.77 +/- 0.13) u x mL(-1)] indicating that the variant heavy chain is able to improve the secretion of spliced BDD-FVIII and activity. A cellular mechanism-independent BDD-FVIII splicing was also observed. It provided evidence for ongoing animal experiment using intein-mediated dual-AAV vector technology for delivery of the BDD-FVIII genes.
Factor VIII ; genetics ; metabolism ; secretion ; Glycosylation ; HEK293 Cells ; Humans ; Inteins ; Mutation ; Peptide Fragments ; genetics ; metabolism ; secretion ; Protein Splicing ; Trans-Splicing ; Transfection