Objective To study clone human vascular endothelial growth factor gene165(hVEGF165) to construct the recombined plasmid pcDNA 3.1/hVEGF165 and observe its expression in COS-7. Methods hVEGF gene was amplified by reverse transcription polymerase chain reaction(RT-PCR) method from fetal human myocardium tissue, was then loned into T-vector; identified by polymerase chain reaction(PCR); and was inserted into the expression plasmid pcDNA3.1 to construct the recombined plasmids that encoding VEGF165 comlementary DNA(cDNA). COS-7 cells were transfected mediated by liposome, then expressed protein was detected by Western blotting. Results Exact gene order of hVEGF165 was obtained from the fetal human myocardium tissue by RT-PCR; pcDNA3.1/VEGF165 was constructed, and transient expression was going after transfecting COS-7 cell. Conclusion The recombined plasmids we constructed could successfully express the hVEGF protein after eukaryotic cells COS-7 were transfected.
OBJECTIVE To confirm membrane-guided tissue regeneration in the healing course of segmental bone defects and study the mechanism. METHODS Segmental, 1 cm osteoperiosteal defects were produced in both radii of 12 rabbits. One side was covered with hydroxyapatite/polylactic acid(HA/PLA) membrane encapsulated as a tube. The contralateral side served as an untreated control. Healing courses were detected by radiographic and histologic examinations. RESULTS All control sides showed nonunion, whereas there were consistent healing pattern in test sides. CONCLUSION Membrane technique can promote bone regeneration.
【Abstract】ObjectiveTo construct the eukaryotic expressing plasmid of tumor necrosis factor-related apoptosisinducing ligand（TRAIL），and study its inhibitory effect on hepatic tumor which implanted subcutaneously in nude BALB/c mice.MethodsTotal RNA of U937 cell was extracted, and its extracellular domain (114-281aa) was amplified by RTPCR, then signal peptide was ligated. The recombinant secreting plasmid for TRAIL was constructed successfully which was confirmed by enzyme cleavage identification and sequencing identification. Liver cancer cell (strain No.7402) was implanted subcutaneously in 32 nude BALB/c mice. These mice were randomly divided into two groups: study group and control group. The mice in study group received muscular injection of plasmids for transfection, and the mice in control group received the injection of normal saline at the same time. The size of implanted tumors were measured continuously till the day of sacrificing, tumor cell apoptosis effect was examined by TUNEL method. ResultsIn study group，tumor volume was smaller than that in control group and the bluepurple apoptosis cells were observed under microscope. ConclusionTRAIL plasmid can induce apoptosis of liver cancer cell and can inhibit the growth of liver tumor.
Objective To observe the effect of BMSCs on the cardiac function in diabetes mellitus (DM) rats through injecting BMSCs into the ventricular wall of the diabetic rats and investigate its mechanism. Methods BMSCs isolated from male SD rats (3-4 months old) were cultured in vitro, and the cells at passage 5 underwent DAPI label ing. Thirty clean grade SD inbred strain male rats weighing about 250 g were randomized into the normal control group (group A), the DM group (group B), and the cell transplantation group (group C). The rats in groups B and C received high fat forage for 4 weeks and the intraperitoneal injection of 30 mg/kg streptozotocin to made the experimental model of type II DM. PBS and DAPI-labeledpassage 5 BMSCs (1 × 105/μL, 160 μL) were injected into the ventricular wall of the rats in groups B and C, respectively. After feeding those rats with high fat forage for another 8 weeks, the apoptosis of myocardial cells was detected by TUNEL, the cardiac function was evaluated with multi-channel physiology recorder, the myocardium APPL1 protein expression was detected by Western blot and immunohistochemistry test, and the NO content was detected by nitrate reductase method. Group C underwent all those tests 16 weeks after taking basic forage. Results In group A, the apoptosis rate was 6.14% ± 0.02%, the AAPL1 level was 2.79 ± 0.32, left ventricular -dP/dt (LV-dP/dt) was (613.27 ± 125.36) mm Hg/s (1 mm Hg=0.133 kPa), the left ventricular end-diastol ic pressure (LVEDP) was (10.06 ± 3.24) mm Hg, and the NO content was (91.54 ± 6.15) nmol/mL. In group B, the apoptosis rate was 45.71% ± 0.04%, the AAPL1 level 1.08 ± 0.24 decreased significantly when compared with group A, the LVdP/ dt was (437.58 ± 117.58) mm Hg/s, the LVEDP was (17.89 ± 2.35) mm Hg, and the NO content was (38.91±8.67) nmol/mL. In group C, the apoptosis rate was 27.43% ± 0.03%, the APPL1 expression level was 2.03 ± 0.22, the LV -dP/dt was (559.38 ± 97.37) mm Hg/ s, the LVEDP was (12.55 ± 2.87) mm Hg, and the NO content was (138.79 ± 7.23) nmol/ mL. For the above mentioned parameters, there was significant difference between group A and group B (P lt; 0.05), and between group B and group C (P lt; 0.05). Conclusion BMSCs transplantation can improve the cardiac function of diabetic rats. Its possible mechanismmay be related to the activation of APPL1 signaling pathway and the increase of NO content.
【摘要】 目的 探讨宫颈上皮内瘤变（CIN）和宫颈鳞癌（SCC）组织中人类染色体端粒酶基因（hTERC）的表达和临床意义。 方法 收集2007年10月-2009年6月经病理学证实的116例宫颈脱落细胞标本，其中LSIL（CINⅠ）30例、HSIL（CINⅡ/Ⅲ）37例、SCC 16例、宫颈炎33例，用荧光原位杂交（FISH）方法检测脱落细胞hTERC基因。 结果 在宫颈炎、LSIL、HSIL和SCC组中hTERC基因的表达率分别是6.1%、16.7%、51.4%和93.8%，其中，HSIL、SCC组与宫颈炎组比较，hTERC基因阳性率差异有统计学意义（Plt;0.05），LSIL组与HSIL组比较、LSIL组与SCC组比较、HSIL组与SCC组比较，差异有统计学意义（Plt;0.05），且随着病变程度增加，hTERC基因表达率增加。 结论 hTERC基因在细胞学LSIL、HSIL和SCC中表达异常，且随病变程度增加阳性表达率也增加，可作为宫颈癌癌前病变进展的生物遗传学监测指标，并有望成为宫颈癌早期筛查方法之一。【Abstract】 Objective To explore the clinical significance and expression of the human telomerase gene (hTERC) in the cervical intraepithelial neoplasia (CIN) and squamous carcinoma of the cervix (SCC). Methods According to histological biopsy from October 2007 to June 2009, 116 pap smears were divided into LSIL (n=30), HSIL (n=37), SCC (n=16), and cervicitis (n=33) groups. Fluorescence in situ hybridization (FISH) was used to detect the expression of hTERC. Results Positive expression rate of hTERC was 6.1% in cervicitis group, 16.7% in LSIL group, 51.4% in HSIL group, and 93.8% in SCC group, respectively. Compared to cervicitis group, the expression of hTERC in HSIL and SCC groups was significantly higher (Plt;0.05). Among LSIL, HSIL, and SCC groups, there were significant differencec in hTERC expression between every two groups (Plt;0.05). From LSIL to SCC, the expression of hTERC increased obviously. Conclusion Abnormal expression of hTERC exists in LSIL, HSIL, and SCC patients, which significantly increases during malignant development. It may be a biogenetics monitor index of cervical precancerosis and will be a screening marker for cervical cancer.
ObjectiveTo explore the epidemiological characteristics and analyze the related diagnosis and treatment information of wound outpatient cases in a tertiay hospital in Leshan City.MethodsThe data of cases admitted to the wound clinic in a tertiary hospital of Leshan City between January 1st, 2017 and December 31st, 2019 were retrospectively collected. The SPSS software was used for statistical analyses in terms of basic patient information, wound type and dressing use, number of visits and expenses.ResultsA total of 10 880 cases were enrolled. The average age of the patients treated was (45.23±13.45) years old. Patients from the Central District, where the hospital was located in, accounted for the highest proportion (38.63%). The proportion of acute skin and soft tissue injuries was higher than that of chronic skin and soft tissue injuries (62.51% vs. 37.49%). According to the proportions from high to low, the top three chronic skin and soft tissue injuries were postoperative refractory wounds (19.63%), skin and soft tissue infections (3.90%), and stoma complications (3.04%), respectively. A total of 22 937 functional wound dressings were used, of which non-woven outer dressings accounted for 47.52%. Patients with acute skin and soft tissue injuries had a higher healing rate compared with those with chronic skin and soft tissue injuries (96.65% vs. 43.03%, P<0.05). The type of skin and soft tissue injuries with the most visits was cancerous wound [the median (lower quartile, upper quartile) was 68 (35, 121) visits], and the one with the least visits was acute skin and soft tissue injuries [the median (lower quartile, upper quartile) was 2 (1, 7) visits].ConclusionsAs the distance between residence and hospital grows, the number of patients decreases, suggesting that the specialized nursing assistance has achieved initial results in remote areas, but the task of assistance in surrounding areas is still heavy. The type of skin and soft tissue injuries is changing under the concept of enhanced recovery after surgery, and wound care plays an important role in the recovery of patients after surgery. The concept of wet healing and new dressings to promote wound healing are widely used, but still need to be further popularized among patients and medical staff. For chronic wounds that are more expensive to treat, it is necessary to formulate comprehensive treatment plans such as wound treatment, psychological support, and systemic nutrition to promote healing.
ObjectiveTo systematically review the predictive value of ezrin expression in bone and soft tissue sarcomas.MethodsDatabases including PubMed, EMbase, The Cochrane Library (Issue 2, 2016), CNKI, CBM, VIP, and WanFang Data were searched to collect cohort studies about the prognostic value of ezrin expression in bone and soft tissue sarcomas from inception to May 2016. Two reviewers independently screened literature, extracted data and assessed the risk of bias of included studies. Then meta-analysis was performed by using RevMan 5.3 software.ResultsFinally, a total of 17 cohort studies, involving 1 460 patients were included. The results of meta-analysis showed that: the overall survival (HR=1.90, 95%CI 1.70 to 2.13, P<0.000 01), event-free survival (HR=2.42, 95%CI 1.53 to 3.84, P=0.0002), metastasis-free survival (HR=2.09, 95%CI 1.10 to 3.97, P=0.02) in the bone and soft tissue sarcomas patients with ezrin high expression were lower than patients with lower expression. The same results were also observed in subgroup analysis according to histologica type and ethnicity in overall survival.ConclusionEzrin high expression may be used as a predictive maker for poor prognosis in bone and soft tissue sarcomas. For the quantity and quality limitation of the included studies, this conclusion still needs to be further proved by performing more high quality studies.
ObjectiveTo evaluate the efficacy and safety of the third-generation super-mini percutaneous nephrolithotomy (SMP) for the treatment of kidney calculi with diameter of ≤2.5 cm and CT value of ≥700 Hu, and discuss the feasibility of adopting the technology in primary hospitals.MethodsThe clinical data of 64 patients with unilateral kidney calculi (CT value ≥700 Hu, diameter ≤2.5 cm) treated in the People’s Hospital of Leshan Central District between July 2017 and July 2018 were retrospectively analyzed. After random assignment, 30 patients were treated with SMP and 34 were with mini percutaneous nephrolithotomy (MPCNL). The pre-, intra-, and post-operative data were compared and analyzed to evaluate the efficacy and safety.ResultsThe unilateral lesion operations of both groups were successfully completed in the first phase. All patients were given double J tubes after operation, and there were no major complications such as post-operative hemorrhage and sepsis. There was no statistically significant difference in the post-operative hemoglobin decrease, post-operative immediate stone removal rate, post-operative stone removal rate after one month, or the rate of procalcitonin >0.1 μg/L between the two groups (P>0.05). The differences in the lithotripsy time [(29.63±6.28) vs. (25.21±5.19) minutes], post-operative hospital stay [(5.33±0.61) vs. (9.44±0.96) days], rate of indwelling renal fistula (3.3% vs. 50.0%), analgesic demand rate (10.0% vs. 58.8%), and postoperative infectious fever rate (6.7% vs. 26.5%) between SMP group and MPCNL group were statistically significant (P<0.05).ConclusionsSMP has the advantages of less trauma, low systemic inflammatory response syndrome incidence, less pain, quick rehabilitation, short hospital stay, tubeless after surgery, etc. It is worthy of extensive promotion in primary hospitals.
Objective To establish chronic hindlimb ischemia model with suture-occluded method in rats， and then compare the effects of chronic hindlimb ischemia model with acute ischemia model. Methods Models of chronic hindlimb ischemia were established by using suture-occluded femoral artery method. The laser Doppler blood flow analysis and angiography were performed on day 7， 14， 28， 42， and 49 after operation， and then the rats were sacrificed after angiography， respectively， the quadriceps and gastrocnemius of contralateral and ipsilateral （surgical side） were gotten， which were tested by HE staining and α-actin immunohistochemistry staining， and then calculate arteriolar density. Results There were no lameness and limb necrosis after operation in chronic hindlimb ischemia models. Laser Doppler analysis found that chronic hindlimb ischemia models were still maintained in ischemia state on day 49 after operation compared with acute ischemic models. The resluts of HE staining showed no acute necrosis and muscle fibrosis in chronic hindlimb ischemia model group. Chronic hindlimb ischemia models after operation did not appear obvious lameness and limb necrosis. The arteriolar density of quadriceps femoris on day 7 after operation in chronic hindlimb ischemia models were less than that in acute hindlimb ischemia models （0.015 2 vs. 0.036 4）. Conclusions Compared with the commonly used acute ischemic models， the duration of arterial limb ischemia in chronic hindlimb ischemia rats， which were established by suture-occluded method， is longer and less likely to be affected by the compensatory mechanisms. So suture-occluded method can provide a new animal hindlimb ischemia model for further study of ischemia angiogenesis mechanism and treatment of severe lower extremity ischemia.
Objective To investigate the feasibil ity of preparing the porous extracellular matrix (ECM) by use of some chemicals and enzymes to decellularize the porcine carotid artery. Methods The porcine carotid artery was procured, and warm ischemia time was less than 30 minunts. The porcine carotid artery was decellularized with 1% sodium dodecyl sulfate (SDS) for 60 hours to prepare common ECM; then common ECM was treated with 0.25% trypsin (for 6 hours) and 0.3 U/ mL collagenase (for 24 hours) to prepare porous ECM. The common ECM and porous ECM were stained with HE,Masson’s trichrome, and Orcein to evaluate the histological features. Then the mechanical property, cytotoxicity, and pore size of ECMs were determined. After 4 weeks of subcutaneous implantation in dogs, the histological examination was used for the study. Results Histological observation confirmed that 2 kinds of ECMs were decellularized completely and more porous structure was observed in porous ECM. Scanning electron microscope showed the pores in porous ECM were greater and the length of shorter axis in porous ECM ranged from 5 to 30 μm, the length of longer axis from 40 to 100 μm. The porosity of porous ECM (99.25%) was greater than that of common ECM (91.50%). The burst pressure of porous ECM decreased when compared with common ECM, showing significant difference [(0.154 3 ± 0.012 7) MPa vs [0.305 2 ± 0.015 7) MPa, P lt; 0.05]. There was no significant difference in suture retention strength between 2 kinds of ECMs (P gt; 0.05). The cytotoxicity test showed no obvious cytotoxicity in 2 kinds of ECMs. In vivo implantation test showed that the deeper host cells infiltration and more neo-microvessels in porous ECM were observed than in common ECM. Conclusion SDS and some enzymes can be used to prepare porous ECM as the scaffold for tissue engineered blood vessels.