Experimental study of curcumol inhibiting vascular endothelial growth factor-induced neovascularization / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To study the role and mechanism of curcumol in neovascularization induced by vascular endothelial growth factor(VEGF).METHODS: Human umbilical vein endothelial cells were cultured in vitro and treated with 50ng/mL VEGF and curcumol at different concentrations. Cell proliferation was detected by CCK-8 and EdU assay, the migration ability of cells was analyzed by Transwell assay, the angiogenesis ability of endothelial cells was analyzed by tube formation assay, and the change of Akt/mTORC1 signal pathway was detected by Western blot.RESULTS: CCK-8 results showed that the OD450 value of cells in 400 and 800 μmol/L curcumol+VEGF group was significantly lower than that in VEGF group(all P<0.01). EdU results showed that the rate of cell proliferation in 400 μmol/L curcumol+VEGF group was significantly lower than that in VEGF group(P<0.001). Transwell assay and the formation assay results showed that the number of migratory cells in 400 μmol/L curcumol+VEGF group was decreased, and the number and length of tube branches were also reduced compared with VEGF group(all P<0.001). Western blot results showed that curcumol significantly inhibited the expression of p-Akt and p-S6, which were downstream targets of Akt/mTORC1 pathway in cells.CONCLUSION: Curcumol can inhibit VEGF-induced cell proliferation, migration and tube formation of vein endothelial cells, and has a strong inhibitory effect on angiogenesis, which can be further studied in the treatment of ocular fundus neovascularization.

Role of p53 Protein in Improving Nerve Function of Rats with Acute Cerebral Ischemia-reperfusion Injury by Electroacupuncture Pretreatment / 中国康复理论与实践

@#Objective To investigate the effect of p53 protein on the nerve function of rats with acute cerebral ischemia-reperfusion by electro-acupuncture pretreatment. Methods A total of 72 male Sprague-Dawley rats were randomly divided into sham group(n=24),model group(n=24) and electroacupuncture pretreatment group(n=24).Each group was equally divided into two hours and 72 hours subgroups. The neurological behavior was assessed with modified Neurological Severity Score (mNSS). HE staining and TUNEL assay were used to observe the injury and cell apoptosis in ischemic brain,and Western blot-ting was used to detect the expression of p-p53(ser392),p53 and microtubule associated protein 1 light chain 3 (LC3)II in ischemic area. Results Compared with the model group,the mNSS scores,the injury of brain tissue,the number of TUNEL-positive cells, and the level of p-p53 and LC3II decreased (P<0.05) in the electroacupuncture pretreatment group.Two hours after reperfusion,there was no significant difference in the level of p53 between the model group and the electroacupuncture pretreatment group(P>0.05),however,72 hours after reperfusion,the level of p53 decreased in the electroacupuncture pretreatment group(P<0.05). Conclusion Electroacupuncture pretreatment can improve neurological deficit in cerebral ischemia-reperfusion injury rats,which may be related to the involvement of p53 protein in autophagy and apoptosis.

Taohong Siwu Decoction regulated functions of endothelial cells and treated arteriosclerosis obliterans: an experimental study / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To discuss the effect of Taohong Siwu Decoction (TSD) in regulating functions of endothelial cells and treating arteriosclerosis obliterans (ASO).</p><p><b>METHODS</b>The ASO model was prepared by using high-fat diet plus intimal injury. They were randomly divided into the model group (n = 10), the normal control group (n = 9), the low dose TSD group (group A, n = 12), the middle dose TSD group (group B, n = 10), and the high dose TSD group (group C, n = 9). Eight weeks after modeling, the limb blood perfusion was observed using laser Doppler flowmetry. The arterial morphology was observed using light microscope and transmission electron microscope. The number of circulating endothelial cells (CECs) was determined using Percoll density gradient centrifugation method. Serum levels of TNF-alpha, IL-1, ET-1, and NO were detected using double antibody sandwich assay of enzyme linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>The ASO rat model was successfully established. Blood lipids levels significantly increased, the blood perfusion of left hind limbs significantly decreased, the number of CECs in the peripheral blood significantly increased, the arterial lumen was irregularly narrowed, the ultra-structure of vessel walls was damaged, serum levels of TNF-alpha, IL-1, and ET-1 significantly increased, and the serum level of NO significantly decreased in the model group, showing statistical difference when compared with the normal control group (P < 0.01). Compared with the model group, significant improvement in the aforesaid indices was shown in group B and C (P < 0.05, P < 0.01).</p><p><b>CONCLUSIONS</b>The injury and abnormal functions of endothelial cells is an important pathological process of ASO. As an effective recipe for treating ASO, TSD could protect vascular endothelial cells and improve the secretion function of vascular endothelial cells.</p>

Ultrasound bio-microscopy for measurement of coronary artery flow and estimation of infarct size in a mouse model of acute myocardial infarction.

Ultrasound bio-microscopy was used to measure hemodynamic changes in the left main coronary artery after myocardial infarction (MI), and its usefulness in estimating infarct size was evaluated. MI was induced by left anterior descending artery ligation. Diastolic peak velocity (Vd), mean flow velocity (Vmean) and the velocity-time integral (VTI) were measured 2 and 6 h after MI. Serum troponin I levels were assayed 2, 6 and 12 h after MI. At 2 h, Vmean and VTI significantly differed between mice that underwent low and high left anterior descending artery ligation; Vd, Vmean and VTI were correlated with infarct size (r = -0.557, -0.693 and -0.672, respectively; all p < 0.01). Infarct size was more strongly correlated with 2-h ultrasound bio-microscopy measurements than with 2-h serum troponin I level. Measurement of coronary artery blood flow by ultrasound bio-microscopy may be useful for early estimation of infarct size in mice.

Relationship between bronchoalveolar lavage mycoplasma load and clinical characteristics in children with Mycoplasma pneumoniae pneumonia / 中华儿科杂志

<p><b>OBJECTIVE</b>Mycoplasma pneumoniae (MP) is an important pathogen for community-acquired pneumonia in children. MP infection was considered to be self-limited, but many severe refractory MP pneumonia cases have been reported in recent years. The reason for variation in severity of MP pneumonia remains unclear. MP virulence including drug-resistance and host immunologic function are important influencing factors. The present study aimed to clarify relationship between local MP load and severity of MP pneumonia.</p><p><b>METHOD</b>MP DNA was quantitatively detected by fluorescent real-time PCR in bronchoalveolar lavage fluid (BALF) from 77 children with MP pneumonia. They were classified into groups of low MP load ( < 10(3)/ml, n = 14) , moderate MP load (10(3)-10(6)/ml, n = 22) and high MP load ( > 10(6)/ml, n = 41) . Clinical symptoms, main laboratory and imaging results of children among the three groups were compared.</p><p><b>RESULT</b>When compared with low load group and moderate load group, high load group had longer fever duration (7 d, 10 d vs. 12 d) , longer time to normalization of temperature with macrolide administration (4 d, 8 d vs. 10 d) , more patients with high fever (50.0%, 68.2% vs. 87.8%) and longer duration of fever than 10 d (35.7%, 50.0% vs. 73.2%).Statistically significant difference existed in CRP among the three groups (1.0 mg/L, 11.5 mg/L, 34 mg/L). Large field of consolidation or atelectasis were found in 58.5% of high load patients, much higher than 22.7% in moderate load and 14.3% in low load patients. Bilateral or massive pleural effusion was not found in low load group, while in moderate load and high load group, they were 13.6% and 24.4%. However, no significant difference was found in symptoms and main laboratory and imaging results among different age groups in high load patients.</p><p><b>CONCLUSION</b>There is a close relationship between MP load in BALF and clinical characteristics in children with MP pneumonia. Those with high MP load have a more severe process.</p>

Screening for EXT1 and EXT2 gene mutations in a ethnic Han Chinese family from Shanxi with hereditary multiple exostoses / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To screen for potential mutations in an ethnic Han Chinese family from Shanxi with hereditary multiple exostoses.</p><p><b>METHODS</b>Polymerase chain reaction and DNA sequencing were used to screen potential mutations in EXT1 and EXT2 genes.</p><p><b>RESULTS</b>For EXT1 gene, two synonymous mutations (P477P and E587E), three intronic mutations (c.1537 -48A>G, c.1721 +203A>G and c.1722 -103C>G) were detected. For EXT2 gene, five intronic mutations (c.-29 -148A>T, c.1080 -18T>A, c.1336 -93C>T, c.1526 -166C>T, and c.1526 -195C>T) were identified. Among these, EXT1 P477P, EXT1 E587E and EXT2 c.1080 -18T>A are polymorphisms listed by Multiple Osteochondroma Mutation Database, whilst the other 7 sites have not been reported.</p><p><b>CONCLUSION</b>No mutations have been found among all exons of the EXT1 and EXT2 genes in this family. Linkage analysis is necessary for identifying the cause of this disease.</p>

Mixed infections in children with Mycoplasma pneumoniae pneumonia / 中华儿科杂志

<p><b>OBJECTIVE</b>To investigate the incidence and clinical features of mixed infections in children with Mycoplasma pneumoniae (MP) pneumonia.</p><p><b>METHOD</b>A total of 201 cases diagnosed as MP pneumonia were investigated for mixed infections by sputum bacterial culture, respiratory virus antigen detection and serum Chlamydia pneumoniae antibody test. For those with the indications for bronchoscopy, we also did bronchoalveolar lavage and lavage bacterial culture.</p><p><b>RESULT</b>A high incidence (103/201, 51.2%) of mixed infections in children with MP pneumonia was revealed. The most frequent co-infected pathogen was Chlamydia pneumoniae (52, 25.9%), followed by viruses (29, 14.4%), and bacteria (22, 10.9%). Among viruses, respiratory syncytial virus was the most common (17, 8.5%), followed by adenovirus (6, 3.0%), parainfluenza virus type III (4, 2.0%) and influenza virus type B (2, 1.0%). Sputum bacterial culture was positive in 14/201 (7.0%) cases, Streptococcus pneumonia being most common (6, 3.0%). BALF culture yielded positive results in 11.6% (8/69), Streptococcus pneumonia was also common (5, 7.3%). Among 29 cases with MP and virus coinfection, 26 were younger than 3 years (89.7%), while for MP and Chlamydia pneumoniae coinfection, most of them were older than 3 years (40/52, 76.9%). Compared with non-mixed infections, those with mixed infections had longer fever duration (24.5% and 40.8% longer than 10 d), more frequently developed pleural effusion (11.2%, 23.3%) and large area of shadow in chest imaging (35.7%, 51.5%). White blood cell [(14.28 ± 4.99) × 10(9)/L], C-reactive protein (CRP) [69(32.5 - 99.5) mg/L] and neutrophil ratio in BALF [0.86 (0.63 - 0.91)] were much higher in children with mixed bacterial infections than that in non-mixed infections [(9.06 ± 3.47) × 10(9)/L, 3 (0 - 31.0) mg/L, 0.44 (0.03 - 0.88)]. But no significant difference was found in peripheral blood neutrophil proportion between mixed bacterial infections (0.38 ± 0.25) and non-mixed infections (0.51 ± 0.19).</p><p><b>CONCLUSION</b>More than half of cases with MP pneumonia had mixed infections, most commonly caused by Chlamydia pneumonia followed by viruses. The incidence of mixed infections with bacteria was low. Mixed infections with virus were more common in young children, while mixed infection with Chlamydia pneumoniae was more common in older ones. Bacterial infections should be paid more attention, especially those caused by Streptococcus pneumoniae, for those with high peripheral white blood cell counts, high CRP levels and high proportion of neutrophils in BALF.</p>

Influence of Chinese drugs for cooling blood and dissolving stasis in different concentrations on morphology of choroidal neovascularization / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To evaluate the influence of Chinese drugs for cooling blood and dissolving stasis (CBDS) in different concentrations on morphology of krypton laser induced choroidal neovascularization (CNV) in brown Norway (BN) rats.</p><p><b>METHODS</b>Forty-eight rats received laser irradiation (659 nm) on fundus of one eye (power 360 mW, spot diameter 50 microm, time 0.05 s). They were divided into four groups equally: the control group (A) treated with normal saline, and the three CBDS groups treated respectively with high (B, 5.0 g/kg), median (C, 2.5 g/kg) and low (D, 1.25 g/kg) dosage of CBDS, twice every day via gastric perfusion for 21 successive days. Fundus fluorescein angiography (FFA) on 4 selected rats in each group was performed at the 7th, 14th and 21st day after photocoagulation, and histopathologic examination using light microscope with immuno-histochemical stain was conducted on them as well.</p><p><b>RESULTS</b>FFA showed that CNV was firstly appeared on day 7 after photocoagulation, in Group A, it expanded gradually and reached the peak on day 21, but showed no significant expansion in the three CBDS groups. The fluorescein leakage in Group C (52343.13 +/- 12973.92 dots) and D (66252.78 +/- 20659.71 dots) was significantly less than that in Group A (91457.19 +/- 29309.11 dots) and B (95973.40 +/- 53950.43 dots) on day 21, all showing statistical significance (P<0.05). The variation of CNV in thickness showed that in Group A it increased gradually from day 7 and reached the peak on day 21 (55.3383 +/- 8.5036 microm); but in the CBDS groups, the peak was reached on day 14, then became thinned gradually, on day 21, it was 40.0913 +/- 13.3448 microm in Group B, 38.8473 +/- 7.9862 microm in Group C and 38.9372 +/- 5.1728 microm in Group D, all thinner than that in Group A significantly (P<0.05).</p><p><b>CONCLUSION</b>CDBS can effectively suppress the krypton laser induced CNV proliferation and prevent the CNV leakage in BN rats.</p>

[Establishment of segregating inbred strain of Yuyi hairless mice and its monitoring of genetic characteristics].

Using brother-sister inbreeding with forced heterozygosity to breed a new segregating inbred strain which carry mutant hairless gene. Then, genetic monitoring was conducted by the skin grafting test, coat color test and biochemical marker analysis, and its basic biological characteristics were studied with corresponding methods. The Results are that the Yuyi Hairless Mice (YYHL) with unique biological characteristics have been bred successfully and have progressed to 30th generation since 1991. Thirteen biochemical markers loci on nine chromosomes coding biochemical markers measured with electrophoresis were all homogenous. The skin grafting test showed that no dropping graft was found during 100 days after transplanting, implying the YYHL was of isohistogeneicity. Analysis of coat color genes indicated that the hair color of first generation hybrid crossed between YYHL and DBA/2 was all agouti suggesting that the coat color genes of the YYHL were homogenous. The gene type is AABBccDD. All these showed that the YYHL have been an inbred strain reaching the international standards.