The different biological functions were studied in mouse bone marrow-derived endothelial progenitor cells isolated by differential time attachment to obtain the optimal adherent time in this study. Density gradient centrifugation-isolated bone marrow mononuclear cells were seeded on the fibronectin-coated dish. The 1-day cultured unattached cells were seeded on the second dish for 2 more days. Then unattached cells in the second dish were seeded on the third dish. The cells on 3 dishes were defined as 1-day adherent cells, 3-day adherent cells and 3-day unattached cells, respectively. After 20-day culture, the biological functions, such as the percentage of biomarkers, the ability of adhesion, and the ability of forming tubes in vitro were analyzed. The results showed that the percentages of positive CD34, FLK-1, and CD34/FLK-1 expressions in 1-day attached cells were significantly increased compared to those in the 3-day adherent or unattached cells (P < 0.01), which showed the strongest adhesion ability. The expression of eNOS in 1- or 3-day adherent cells was significantly higher than that in 3-day unattached cells (P < 0.01). The expression of VEGF in 3-day adherent cells was significantly higher than that in 1-day adherent cells or 3-day unattached cells (P < 0.01). These results suggest the biological functions of 1-day adherent cells are significantly stronger than that of 3-day adherent or unattached cells. VEGF expression in 3-day adherent cells is higher than that in 1-day adherent cells or 3-day unattached cells. The expression of eNOS in 1-day adherent cells or 3-day adherent cells is higher than that in 3-day unattached cells. The optimal adherent time to obtain mouse bone marrow-derived endothelial progenitor cells is 1-3 d.
Animals ; Bone Marrow Cells ; cytology ; Cell Culture Techniques ; methods ; Cell Differentiation ; Cell Separation ; methods ; Cells, Cultured ; Endothelial Cells ; cytology ; metabolism ; Leukocytes, Mononuclear ; cytology ; Male ; Mice ; Mice, Inbred C57BL ; Nitric Oxide Synthase Type III ; metabolism ; Stem Cells ; cytology ; metabolism ; Time Factors ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo discuss the curative effect of the external fixator for complex tissue defect in the forearm.

METHODSFrom May, 2005 through December, 2008, the external fixators were used in 17 patients to treat the complex tissue defect in the forearm caused by trauma. There were 11 male and 6 female, with a mean age of 25.6. All patients were accompanied with the exposure of tendon, muscle or screw. The skin defect ranged from 7 cm x4 cm to 19 cm x9 cm. All patients underwent pedicle flap repair. The flap ranged from 10 cm x 6 cm to 20 cm x 15 cm. The proximal pedicle of the flap was sutured into a tubular. The position of the pedicle was fixed by the external fixator. The pin was at the ulnar and the iliac (n=5), and the radius and the iliac (n=12). The immobilization lasted 3 to 8 weeks, 5.1 weeks in average.

RESULTSAll patients were followed up for 3 to 20 months, 11.3 in average. All pedicle flaps survived with no pressure ulcer, or no erosion in the axilla. No compartment syndrome or osteomyelitis occurred. Four to six week after surgery, the pedicle was cut. Infection occurred at the cutting end in 1 patient. The wound healed after addressing. The wound in the other 16 patients healed successfully. The fracture of the ulnar and the radius healed 8.5 or 15 weeks after surgery, 13.5 weeks in average. Eleven patients underwent second stage reshape and function restoration. The function of the hands and forearms recovered satisfactorily. Eleven patients returned to their work. Six patients can live with basic function for living.

CONCLUSIONSThe external fixator used for complex tissue defect in the forearm can keep the position of the pedicle, replacing plaster fixation. It can reduce the incidence of flap and vessel spasm, and get good outcomes.

Adult ; External Fixators ; Female ; Follow-Up Studies ; Forearm Injuries ; surgery ; Humans ; Male ; Middle Aged ; Surgical Flaps ; Young Adult

OBJECTIVETo investigate the specificity and sensitivity of the immunohistochemistry for hMLH1 and hMSH2 with detection of microsatellite instability (MSI) to identify the kindreds with hereditary nonpolyposis colorectal cancer and to analyse its value in clinical practice.

METHODSpecimens of 16 cases with HNPCC and 16 cases with sporadic colorectal cancer were detected by immunostaining with hMLH1 and hMSH2 and MSI was also detected.

RESULTSThe specificity and sensitivity of the immunohistochemistry for hMLH1 and hMSH2 were 91.7% and 87.5% respectively. The specificity and sensitivity of MSI were 100% and 75.0%. By combining two methods, the specificity and sensitivity were 91.7% and 93.8% respectively.

CONCLUSIONSBy combination of the immunohistochemistry for hMLH1 and hMSH2 and detection of MSI to identify the kindreds with HNPCC, the specificity and sensitivity are improved which is better than to use either of them alone. And it is very easy and cheap that it can be used in clinics.

Adaptor Proteins, Signal Transducing ; Adult ; Aged ; Carrier Proteins ; Colorectal Neoplasms, Hereditary Nonpolyposis ; genetics ; DNA-Binding Proteins ; analysis ; Female ; Genomic Instability ; Humans ; Immunohistochemistry ; Male ; Microsatellite Repeats ; Middle Aged ; MutL Protein Homolog 1 ; MutS Homolog 2 Protein ; Neoplasm Proteins ; analysis ; Nuclear Proteins ; Proto-Oncogene Proteins ; analysis ; Sensitivity and Specificity

OBJECTIVETo investigate the pathogenesis of ischemic-type biliary lesions (ITBLs) in post-liver transplant patients and the possible therapeutic mechanisms of sirolimus.

METHODSThe clinic data of 32 post-liver transplant patients with ITBLs from May 2004 to December 2010 was analyzed. There were including 25 male and 7 female patients with a median age of 46 years (ranging from 19 to 61 years). Patients were divided into those who received sirolimus (sirolimus group) and those who did not (control group). The expression of IL-2, FoxP3, and IL-10 in the portal area, liver function indexes, and bile duct injury score were assessed pre-ITBL, when ITBLs were identified, and after 6 months of sirolimus treatment.

RESULTSCompared with pre-ITBL optical density (OD) values, there was a significantly increase in IL-2 OD(0.138 ± 0.050 in control group and 0.141 ± 0.052 in sirolimus group), but not FoxP3 and IL-10 OD in both groups at the time ITBLs were diagnosed. After 6 months of treatment, the IL-2, FoxP3, and IL-10 OD values in the control group were not different from those when ITBLs were diagnosed. There was a significant reduction in post-therapy IL-2 OD(0.107 ± 0.043, t = 2.087, P = 0.044), and a significant elevation in FoxP3(0.213 ± 0.039) and IL-10 OD(0.187 ± 0.048) in sirolimus group as compared with those when ITBLs were diagnosed(t = -3.822 and -4.350, both P < 0.01). There was a significant increase in serum levels of ALT, AST, total bilirubin, γ-glutamyl transpeptidase and ALP at the time ITBLs were diagnosed compared with pre-ITBL levels in both groups. After 6 months of treatment, the above indexes had not changed in the control group, but significantly improved in the sirolimus group, and the bile duct injury score in the sirolimus group had significantly decreased(4.4 ± 2.4, Z = -2.568, P = 0.010). The 1-year and 3-year graft survival rates in the control group were 6/13 and 5/13, respectively, and 17/19 and 13/19, respectively, in the sirolimus group (χ(2) = 7.166, P = 0.007; χ(2) = 5.398, P = 0.020, respectively).

CONCLUSIONSSirolimus can downregulate IL-2 expression and upregulate FoxP3 and IL-10 expression, thereby stimulating FoxP3+ Treg cells, suppressing immunopathological damage, and promoting epithelial repair in bile ducts.

Adult ; Bile Duct Diseases ; drug therapy ; Female ; Forkhead Transcription Factors ; metabolism ; Gene Expression Regulation ; drug effects ; Humans ; Interleukin-10 ; metabolism ; Interleukin-2 ; metabolism ; Ischemia ; diet therapy ; Liver Transplantation ; Male ; Middle Aged ; Postoperative Complications ; drug therapy ; Sirolimus ; therapeutic use ; Young Adult

Objective To explore the effect of blocking of gap junction communication on neuronal apoptosis in rabbit hypothalamus after explosive brain injury. Methods Forty-eight rabbits were equally randomized into control group and experimental group. Octanol, a gap junction blocker, was injected through the left ventricle to block the gap junction communication in the experimental group and physiological saline in the control group instead. Rabbit models with explosive brain injury were induced by paper detonators 2 h after that. One, 3 and 7 d after the inducement, these rabbit models were sacrificed and thus, they were subdivided into 3 groups (n=8). The protein expression of gap junctional connexins 43 (Cx43) was observed by Western blotting and caspase-3 protein expressions in the hypothalamus were detected by immunohistochemical staining and RT-PCR. Results The protein expression of Cx43 in the experimental group was significantly lower than that in the control group at all time points (P<0.05), and that in the experimental group reached the peak level 3 d after the brain injury.The expressions of caspase-3 in the experimental group were significantly lower than those in the control group after the injury (P<0.05). Conclusion Blocking of gap junction communication can reduce the incidence rate of neuronal apoptosis in the hypothalamus after brain injury, and Cx43 might play an important regulation role in this process.

OBJECTIVETo evaluate the feasibility and efficiency of embolization of spinal dural arteriovenous fistula (SDAVF).

METHODSFrom December 2010 to May 2012, there were 104 cases of SDAVF were treated, and 26 cases were selected to be treated with embolization. The inclusion criteria was as follows: (1) No anterior or posterior spinal artery originated from the fistula segment; (2) The segmental artery can be catheterized with guiding or micro catheter; (3) High flow in fistula; (4) Patient's situation was not suitable for surgery or general anesthesia. Among 26 cases, there were 22 male and 4 female patients, the average age was 55.9 years (ranged from 34 to 81 years). The locations of SDAVF were 10 cases in thoracic, 9 in lumbar and 7 in sacral segment. The main symptoms were progressive numbness and weakness in both lower extremities, most cases accompanied with difficulties in urination and defecation. The average history was 17.1 months (from 1 to 156 months). ONYX-18 liquid embolic agent or Glubran-2 surgical glue were used as embolic material. The patients not cured with embolization were treated with surgery in the following 1 - 2 weeks. Follow-up evaluation was done with MRI after 3 months and DSA after 6 months, besides physical examination.

RESULTSFifteen from 26 cases achieved immediate angiographic cure results: 14 in 20 cases which embolized with ONYX-18; only 1 in 6 cases with Glubran-2. Three in 10 cases of thoracic SDAVF and 12 in 16 cases of lumbar/sacral SDAVF were cured with embolization. Partially embolized cases were treated with surgical obliteration of drainage veins within 2 weeks. Cured patients experienced immediate improvement after embolization and kept getting better in the follow-up. All the patients had MRI follow-up after 3 months and DSA follow-up after 6 months. In 6 month's follow-up, MRI showed the edema and flow void signal in the spinal cord disappeared. DSA showed no fistula recurrence or remnant. There was no deterioration case in all of the embolized cases.

CONCLUSIONSParticular SDAVF is suitable for embolization with ONYX-18. Most lesions located in lumbar and sacral segment are good indications for embolization.

Adult ; Aged ; Aged, 80 and over ; Central Nervous System Vascular Malformations ; therapy ; Embolization, Therapeutic ; methods ; Feasibility Studies ; Female ; Humans ; Male ; Middle Aged ; Treatment Outcome

The global patent data on extracorporeal membrane oxygenation(ECMO)in IncomPat Global Patent Database as of August 29,2022 were retrieved.The development trend and layout of ECMO industry were analyzed in terms of global patent application trend,patent distribution,patent technology,major patent applicants and their patent layout.Some suggestions were put forward for the innovation and development of ECMO industry in China so as to provide references for the formulation of national industrial policy,planning of industry technology direction and enterprise technology research and development and patent layout.[Chinese Medical Equipment Journal,2023,44(10):68-75]