BACKGROUND:Recent studies found that some factors play important role in the process of denervated muscle atrophy, especial y the feak-headbox transcription factor, is the key element to regulate the denervated muscle atrophy.
OBJECTIVE:To investigate the effect of RNA interference on inhibiting feak-headbox 3a gene expression in vitro.
METHODS:The myoblast cel line L6 were cultured in the 6-wel cel culture plates, then pEGFP-N1 and smal
interfering RNA recombinant plasmid with the same ratio was transfected under the Lipofectamine2000 mediation to optimize the transfection efficiency of the detection system;2μg smal interfering RNA recombinant plasmid of feak-headbox 3a gene were transfected with myoblast cel line L6 for 48 and 72 hours.
RESULTS AND CONCLUSION:At 48 hours after pEGFP-N1 and siRNA recombinant plasmid transfection, a large number of bright green fluorescent displayed under fluorescence microscope with higher transfection efficiency. Real-time quantitative PCR analysis showed that there were significant differences in the sequences of feak-headbox 3a-Ⅰ, feak-headbox 3a-Ⅱ, feak-headbox 3a-Ⅲ, feak-headbox 3a-Ⅳ on feak-headbox 3a mRNA when compared with the control group at 48 and 72 hours after trasfection (P<0.05), and the inhibition effect was more significant at 72
hours after transfection when compared with that at 48 hours after transfection. Western Blot gray analysis showed that there were significant differences in sequences of feak-headbox 3a-Ⅰ, feak-headbox 3a-Ⅱ, feak-headbox 3a-Ⅲ,
feak-headbox 3a-Ⅳ on feak-headbox 3a mRNA when compared with the control group at 48 and 72 hours after
trasfection (P<0.05), and the inhibition effect was more significant at 72 hours after transfection when compared with that at 48 hours after transfection, which was same with the effect on mRNA level. RNA interference in vitro can
significantly inhibit the fork-head transcription factor feak-headbox 3a gene expression, and the inhibition effect of feak-headbox 3a gene smal interfering RNA recombinant plasmid transfected with the sequence on the mRNA and protein level of feak-headbox 3a is not clear, which can provide new idea for the gene therapy of RNA mediated denervated skeletal muscle atrophy.

Objective To investigate the protective effect of meglumine adenosine cyclosphosp (MAC) on the cerebral ischemia-reperfusion (I/R) injury in rabbits.Methods Twenty four healthy rabbits were randomly divided into control group (n =6),I/R group (n =6),MAC pretreated group (n =6),and MAC treated group (n =6).Cerebral ischemia-reperfusion injury model was made by separating and electrocoagulating vertebral arteries and clipping common carotid arteries in the latter 3 groups after anesthesia.The sham-operated group underwent vessel separation without clipping.L/R group was administered with nothing,while MAC pretreated group with MAC before ischemia,and MAC treated group with MAC just after ischemia.Blood was gathered from jugular vein before ischemia,and 30 min,1 h,and 2 h after reperfusion for testing IL-8,superoxide dismutase (SOD) and malondialdehyde (MDA).The brain tissue slice was observed by optical microscope.Results Compared to control group and before ischemia,the levels of IL-8 and SOD in serum were significantly increased and decreased,and the levels of MDA was significantly increased at 30 min after reperfusion in I/R group; the levels of IL-8 and MDA in serum were significantly increased,and the levels of SOD in serum was significantly decreased at 1 h and 2 h after reperfusion in I/R group.The levels of IL-8 in serum was less at 30 min and 1 h and 2 h after reperfusion in MAC pretreated group than in I/R group.At 1 h and 2 h after reperfusion,the levels of MDA in serum was less and the levels of SOD in serum was higher in MAC pretreated group than in I/R group.At 1 h and 2 h after reperfusion,the levels of IL-8 in serum were less and the levels of SOD in serum were higher in MAC treated group than in I/R group.The levels of MDA in serum were less at 2 h after reperfusion in MAC treated group than in I/R group.Compared to I/R group,pathological change was lighter in the MAC pretreated and MAC treated group.Conclusions MAC has a fine cerebral-protective effect and has no side effect.

With the aging of population, cerebral small vessel disease has attracted more and more attention. A growing body of literature has confirmed that retinal vascular changes can be used as a potential marker for the prediction of cerebral small vessel disease. The retina is recognized as a window into cerebrovascular and systemic vascular conditions. Combining traditional fundus photograph and fundus fluorescein angiography with optical coherence tomography angiography, the retinal vascular system of patients with cerebral small vessel disease can be comprehensively analyzed. This paper summarizes and analyzes the application of retinal angiography technology in different image types of cerebral small vessel disease and makes a review, in order to provide reference for the early diagnosis and prevention of cerebral small vessel disease.

OBJECTIVEProtein of Streptococcus mutans is considered as one of the virulence factors due to its ability to mediate the initial attachment of Streptococcus mutans to tooth surface. In this study, an anticaries DNA vaccine pCIA-P was used to immunize rats. The expression of PAc in different tissues in vivo, specific immune response and protection effects against dental caries were observed.

METHODSPlasmid pCIA-P was injected into rats by two different routs: intramuscular injection (i.m.) and targeted salivary gland immunization (TSG). Immunohistochemistry technique was used to detect the expression of PAc. Gnotobiotic rats were vaccinated with pCIA-P by three different approaches: TSG, intramuscular injection and buccal mucosal injection (i.o.). The specific immune responses were evaluated by ELISA and their anticaries effects were evaluated by Keyes caries scores.

RESULTSPAc was expressed in the sarcoplasm and sarcolemma of muscle fibers and submandibular glands, especially strongly positive in duct regions. The levels of serum specific anti-PAc IgG and salivary specific anti-PAc IgA in TSG immunization and buccal mucosal immunization group were significantly higher than those of other groups. The Keyes caries scores of those two groups were significantly lower than those of other groups.

CONCLUSIONThe plasmid pCIA-P could provoke specific immune responses as a novel immunogen. Mucosal immunization with pCIA-P appears to be an effective genetic immunization method against dental caries.

Animals ; Antibodies, Bacterial ; blood ; Bacterial Proteins ; genetics ; immunology ; Dental Caries ; prevention & control ; Germ-Free Life ; Immunization ; Male ; Membrane Glycoproteins ; Rats ; Rats, Wistar ; Streptococcal Vaccines ; immunology ; Streptococcus mutans ; immunology ; Vaccines, DNA ; immunology

Objective To analyze the factors and prognosis of graft recovery after donation after citizens death (DCD) donor renal transplantation.Methods A retrospective analysis of 67 cases of DCD renal transplantation from August 2012 to September 2015 in our hospital was carried out.According to the stability of renal function after operation,the patients were divided into group A (51cases) with stable renal function,and 16 cases in group B (delayed graft function after operation).The clinical data of two groups including age,gender,cause of death,warm ischemia time,type of dialysis,and application of norepinephrine before operation were collected and analyzed.The related factors of graft function recovery were analyzed.Logistic regression analysis was used to analyze the risk factors of graft functional recovery after operation.The 3-month,6-month,1-year and 18-month survival rate after operation in the two groups was compared.Results The causes of death,the time of ischemia,the type of dialysis before operation,the application of norepinephrine before operation,infants and young donors were related factors of graft function recovery (P ＜ 0.05).Logistic regression analysis showed that cerebral hemorrhage death donor,the long thermal ischemia time,the preoperative hemodialysis and the application of norepinephrine before operation were the risk factors of delayed graft function recovery (P＜0.05).The 3-month,6-month,1-year and 18-month survival rate after operation in group A was higher than that in group B,with the difference being statistically significant (P＜0.05).Conclusion Cerebral hemorrhage death donor,the long thermal ischemia time,the preoperative hemodialysis and the application of norepinephrine before operation were the independent risk factors of delayed graft function recovery.And the prognosis of patients with delayed graft function was poor.Clinical risk should be eliminated or reduced in clinical practice,which can effectively prevent the delayed graft function and further improve the prognosis of the patients.

Objective To analyze the effects of laparoscope-assisted Ivor-Lewis surgery on perioperative stress,immune responses and intestinal barrier function in elderly patients with esophageal cancer.Methods A prospective study including 55 elderly esophageal cancer patients undergoing laparoscope-assisted Ivor Lewis surgery (the treatment group,n =25) and Ivor-Lewis surgery(the control group,n=25) was conducted.The white blood cell count,neutrophil-to-lymphocyte ratio,percentages of CD4 and CD8 cells,CD4/CD8 ratio,C reactive protein (CRP) and D-lactic acid levels were compared between the two groups before and at 1,4 and 7 d after operation.Results The white blood cell count(t =2.689,P =0.010) and neutrophil-to-lymphocyte ratio (t =3.300,P =0.002)were lower in the treatment group than in the control group at 1 d after operation.The percentage of CD4 cells was higher in the treatment group than in the control group at 1 d(t =2.242,P =0.029)and 4 d(t =2.031,P =0.047) after operation.The percentage of CD8 cells was higher in the treatment group than in the control group at 1 d after operation(t =2.041,P=0.046).The CD4/CD8 ratio was higher in the treatment group than in the control group at 1 d(t =2.833,P =0.007)and 4 d(t=2.111,P=0.036)after operation.The CRP level was lower in the treatment group than in the control group at 1 d(t=2.267,P=0.028)and 4 d(t =2.111,P =0.036)after operation.The D-lactic acid level was lower in the treatment group than in the control group at 1 d(t =2.267,P=0.028),4 d (t =7.967,P ＜ 0.01) and 7 d (t =2.541,P =0.014) after operation.Conclusions Laparoscopeassisted Ivor-Lewis surgery has good protective effects on perioperative stress,immune responses and intestinal barrier function in elderly patients with esophageal cancer.

Objective:To explore the surgical indications, techniques and methods of closed reduction and minimally invasive fixation for the treatment of pelvic fractures of Tile C2 and C3, and evaluate the clinical efficacy.Methods:A retrospective analysis of the data of 20 cases with Tile C2 and C3 pelvic fractures treated with closed reduction and minimally invasive fixation from January 2016 to July 2019. There were 7 males and 13 female, with an average age of 35.6±14.6 years (range 12-60 years). The time from injury to operation was 5-30 d, with an average of 19.3±7.1 d. Tile classification of pelvic fracture: 13 cases of C2 type and 7 cases of C3 Type. 2 cases were complicated with ipsilateral or bilateral lumbosacral nerve injury. Classification of nerve injury: 2 cases were partial injury, British Medical Research Council (BMRC) Grade M3. The operation is treated with closed reduction and minimally invasive fixation. First, the side with obvious displacement is fixed on the operating table with a pelvic reduction frame, and the side with less displacement is traction. After reduction, insert S 1 and S 2 sacroiliac screw guide-pin on this side to the contralateral sacral fracture. And then change the traction, fix the reset side on the operating table, change the side with obvious traction displacement, after the reset is ideal, pass the inserted guide-pin through the contralateral sacroiliac joint to the outer iliactable. Then insert the sacroiliac screw. The patients complicated with acetabular fracture were reduced and fixed by the corresponding approach, and the anterior ring was fixed by INFIX. The operation time, intraoperative bleeding volume and postoperative complications were recorded. The quality of fracture reduction was evaluated by Matta's criteria, and the clinical effect was evaluated by Majeed score. Results:All the 20 patients successfully completed the operation. The operation time was 105-210 min, with an average of 167.00±31.21 min. The intraoperative bleeding volume was 30-100 ml, with an average of 82.00±5.36 ml. Postoperative X-ray and CT showed that the fracture was reduced and fixed. According to the Matta's criteria, the reduction quality was rated as excellent in 14 cases, good in 4 case, fair in 2 case, with an excellent and good rate of 90%. Two patients showed symptoms of lateral femoral cutaneous nerve injury without other complications related to surgery. Follow-up for 1 to 4 years, the fractures healed, and the healing time was 6 to 12 weeks. According to the Majeed score, the result was rated as excellent in 18 cases, good in 2 case, with an excellent and good rate of 100%.Conclusion:Closed reduction and minimally invasive fixation for the treatment of pelvic fractures of type C2 and C3, with the characteristics of less damage and good results, will become a trend in the treatment of pelvic fractures.

Objective: Remuzzi scoring system is utilized for assessing the degree of renal tissue damage in donors with hypertensive cerebral hemorrhage and donors with brain trauma after cardiac death. To explore the prognosis of hypertensive cerebral hemorrhage donor kidney in renal transplant recipients. Methods: The kidney donated by DCD between January 1, 2016 to June 1, 2018 were retrospectively reviewed. Pathological biopsy was performed before transplantation and hematoxylin-eosin (HE) staining after sectioning. The degree of renal tissue lesions was evaluated by Remuzzi scoring system. According to the source of donor kidney, they were divided into two groups of donors with heart failure due to hypertensive cerebral hemorrhage (HCH) and those with brain trauma (BT). Both groups of donor kidneys were preserved by low-temperature machine perfusion. The immunosuppressive regimen was identical in both groups. The prognosis of two groups was compared by serum creatinine (Scr) at Month 1/6/12 post-operation and cumulative graft survival rate over a follow-up period of 12-36 months. Results: The renal Remuzzi score of HCH donors was significantly higher than that of BT donors. The maximal creatinine clearance rate was significantly lower than that of BT donors [(86.8±27.8) vs (115.4±23.2) ml/min, P<0.05]. At 1/6/12 months post-transplantation, serum creatinine levels were (76.1±18.5), (72.4±16.2) and (71.4±16.8) μmol/L in BT group and (160.3±33.4), (154.3±32.6) and (146.4±29.1) μmol/L in HCH group. The SCr in BT group at 1/6/12 months was lower than that in HCH group (P<0.05). Kaplan-meier analysis showed no significant inter-group difference in graft survival between two groups over a follow-up period of 12 to 36 months (Log-Rank test, P=0.485). Conclusions No significant difference exists in short-term survival rate of kidneys from HCH and BT donors. The recipients of HCH donor's kidney have higher serum creatinine levels than those of BT donors. Selective use of kidney transplants in patients with cardiac death caused by HCH may greatly reduce the waste of donor kidney and improve the quality-of-life of patients with end-stage renal disease.

BACKGROUND: The anti-tumor effect of pigment epithelium-derived factor (PEDF) has been widely confirmed. However, the anti-tumor effect of its peptides is rarely reported. This study aims to investigate the effects of PEDF and its peptides on the apoptosis and migration of non-small cell lung cancer (NSCLC). METHODS: In this study, A549 cells and H1299 cells were selected as the research object, and the cells were divided into normal group, PEDF treatment group, 34 peptide treatment group, 44 peptide treatment group and 34+44 peptide treatment group by administering different drugs at the same concentration to the cells. The proliferation activity of cells in each group was detected by CCK-8 method; the migration ability of cells was detected by scratch test; the expression levels of apoptosis related proteins such as protein kinase 3 (RIP3) and cleaved-caspase-3 were detected by Western blot; the expression levels of epithelial mesenchymal transition (EMT) markers in each group, such as cadherin (E-cadherin) and α-smooth muscle actin (α-SMA) were detected by Western blot; the apoptosis rate of each group was detected by flow cytometry. RESULTS: The results of CCK-8 showed that PEDF and its peptides could inhibit cell proliferation, and the inhibitory effect of 34+44 peptide was the strongest (P<0.05); Observation under the microscope found that PEDF and its peptides can inhibit the proliferation and mesenchymal transformation of A549 cells and H1299 cells, and the inhibitory effect of the 34+44 peptide group is the most obvious; Western blot indicated that compared with other groups, the expressions of cleaved-caspase-3 and RIP3 in 34+44 peptide group were significantly higher (P<0.05), and the expressions of EMT protein E-cadherin were higher, the expression of α-SMA decreased (P<0.05); The results of flow cytometry showed that the apoptosis rate of 34+44 peptide group was significantly higher than those of other groups (P<0.05); The scratch test showed that compared with all the other groups, the healing rate of 34+44 peptide group was the lowest (P<0.05). CONCLUSIONS 34+44 combination peptide can better promote the apoptosis of NSCLC, inhibit the migration of NSCLC, and thereby inhibit the growth of NSCLC.
Apoptosis ; Cadherins/genetics* ; Carcinoma, Non-Small-Cell Lung/genetics* ; Caspase 3 ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Epithelial-Mesenchymal Transition ; Eye Proteins ; Humans ; Lung Neoplasms/genetics* ; Nerve Growth Factors ; Peptides/pharmacology* ; Serpins ; Sincalide

OBJECTIVE: To study the chemical constituents of the roots of Angelica dahurica, a well-known Chinese herbal medicine named Baizhi in Chinese. METHODS: Compounds were separated by various chromatographies, and the structures of new compounds were elucidated based on the analysis of their spectroscopic and spectrometric data (1D, 2D NMR, HRESI MS, IR, and UV). The absolute configurations of new compounds were determined by the calculated electronic circular dichroism and chemical derivatization. The inhibitory activities of all isolates against nitric oxide (NO) production were evaluated using lipopolysaccharide-activated RAW 264.7 macrophage cells. RESULTS: Seven new 3,4-dihydro-furanocoumarin derivatives ( 1a/ 1b, 2a/ 2b, 3a/ 3b, 4) together with a known furanocoumarin ( 5) were isolated from the roots of A. dahurica. The new compounds included three pairs of enantiomers, (4S, 2''R)-angelicadin A ( 1a)/(4R, 2''S)-angelicadin A ( 1b), (4S, 2''S)-angelicadin A ( 2a)/(4R, 2''R)-angelicadin A ( 2b), and (4S, 2''S)-secoangelicadin A ( 3a)/(4R, 2''R)-secoangelicadin A ( 3b), together with (4R, 2''R)-secoangelicadin A methyl ester ( 4). The known xanthotoxol ( 5) inhibited the NO production with the half-maximal inhibitory concentration (IC₅₀) value of (32.8 ± 0.8) µmol/L, but all the new compounds showed no inhibitory activities at the concentration of 100 µmol/L. CONCLUSION This is the first report of the discovery of 3,4-dihydro-furanocoumarins from A. dahurica. The results are not only meaningful for the understanding of the chemical constituents of A. dahurica, but also enrich the reservoir of natural products.