Objective To study the effect and its possible mechanism of p66shc on endothelial dysfunction induced by hydrogen peroxide(H2O2).Methods Human umbilical vein endothelial cells (HUVEC) were used and cultured.H UVEC cells were untreated(control group)or treated with four groups of H2O2,H2O2 plus protein kinase C (PKC) inhibitors,H2O2 plus PKC activator,H2O2 plus P38 inhibitor for 30 minutes respectively,and cells were collected after 24 hours.The apoptosis,viability,proliferation of HUVEC were detected with immune fluorescent staining,MTT and Ki-67 respectively.P66shc and ser36 p66shc (p-p66shc)protein expressions were assessed using Western blotting.P66shc mRNA expression was measured with real-time quantitative polymerase chain reaction (RT-PCR).Results H2O2 induced HUVEC dysfunction which decreased HUVEC proliferation and increased the apoptosis of HUVEC.The expressions of p66shc,p-p66shc protein and p66shc mRNA were significantly increased after treating with H2O2.PKC inhibitor inhibited a H2O2 induced HUVEC dysfunction through increasing HUVEC proliferation activity and reducing cell apoptosis.The expressions of p66shc,p-p66shc protein and p66shc mRNA were significantly decreased after treating with H2O2 plus PKC inhibitor.PKC activator enhanced H2O2-induced HUVEC dysfunction and increased the expressions of p66shc.P38 inhibitor had no obvious effect on H2O2-induced HUVEC dysfunction and the expressions of p66shc.Conclusions p66shc may play an important regulatory rote in endothelial dysfunction caused by H2O2.P66shc may regulate a H2O2-induced endothelial dysfunction through PKC signal pathway.

Objective To observe the therapeutic effect of single lower limb traction plus massage for the lumbosacral tunnel syndrome.Methods The selected 80 candidate patients were divided into the observation group with 38 patients and treatment group with 39 patients. The observation group received single lower limb traction plus massage and the treatment group received pelvic traction plus massage. Both groups therapy last 5 days, The Japanese orthopaedic association scores (JOA) was evaluated for subjective symptoms, signs, functional activity and bladder function score. The therapeutic effect was evaluated.Results The total effective rate (94.7% vs. 82.1%;χ2=12.160,P=0.007) in the observation group was significantly higher than that in the control group. O, The scores of JOA at one week after treatment (20.2 ± 2.0 vs. 18.3 ± 2.8;t=3.419, P<0.01) and two weeks after treatment (24.4 ± 1. 4vs. 22.6 ± 3.0;t=3.359,P<0.01) in the observation group were significantly higher than those in the control group.Conclusions The single lower limb traction plus massage can improve the clinical symptoms of patients with lumbosacral tunnel syndrome, and be superior to the pelvic traction plus massage.

Abstract: Objective　To explore the carrying status of four common deafness genes and mutations on 10 loci in newborns in Hainan, and to analyze the molecular epidemiological characteristics of deafness genes and their loci, so as to provide scientific basis for formulating neonatal deafness gene screening strategy and promoting children's hearing health in Hainan. Methods　Newborns born in Hainan from January 2020 to December 2021 were selected as the research objects. The demographic characteristics of the research objects were collected. At the same time, the plantar blood of newborns was collected, and multiplex PCR amplification and directed hybridization combined with high-throughput sequencing technology were applied to detect 10 mutation loci on 4 common deafness genes. T-test or chi square test was used to process the data. Results　A total of 7 124 newborns were included in the study through informed consent, 219 cases of deafness gene mutation were detected with the detection rate of deafness gene of 3.07%. The detection rates of GJB2, SLC26A4, MT-RNR1 and GJB3 were 1.56% (111/7 124), 1.18% (84/7 124), 0.21% (15/7 124) and 0.11% (8/7 124) respectively. Among the 10 loci of the four genes, the positive detection rate of c.235delC locus of GJB2 was the highest, which was 1.38% (98/7 124), followed by c.919-2A>G of SLC26A4 (0.87%, 62/7 124); 2.63% (113/4 289) of the newborns who passed the preliminary hearing screening still carried the deafness gene; in terms of gene type, the detection rate of GJB2 gene in newborns who failed the hearing screening was higher than that in newborns who passed the hearing screening [2.23% (63/7 124) vs 1.12% (48/7 124)，P<0.01]; in terms of gene loci, the detection rate of c.235delC locus in newborns who failed hearing screening was higher than that in newborns who passed hearing screening [2.09% (59/7 124) vs 0.91% (39/7 124)，P<0.01]. Conclusion　The most common deafness genes types in Hainan were GJB2 and SLC26A4; The most common gene mutation sites were c.235delC and c.919-2A>G; 2.63% of the newborns who passed the preliminary hearing screening still carried the deafness gene, among which the high-risk newborns with MT-RNR1 and GJB3 genes were found. Therefore, hearing screening should be combined with deafness gene screening to improve the detection rate of children at high risk of hearing loss.

OBJECTIVETo study the molecular mechanism of curcumin in human esophageal carcinoma cell line (EC109).

METHODSEC109 cells were cultivated in vitro. When 80%-90% confluence was reached, they were treated with curcumin in different concentrations (15-120 µmol/L). The effects on cell proliferation were examined by CCK-8 colorimetry. The ultrastructure of EC109 cells were detected with transmission electron microscope(TEM). The cells apoptosis was observed with laser confocal microscope(LCM) by AnnexinV-FITC/PI double staining. The proteins level of PTEN, AKT, GSK3β and Caspase 3 were tested by flow cytometry(FCM) .

RESULTSCCK-8 test showed that curcumin could inhibit the proliferation of EC109 cells in a time- and concentration-dependent manner. TEM and LCM examinations indicated that curcumin could make EC109 cells apoptosis. The data of FCM showed that curcumin could increase the expression of PTEN, GSK3β and Caspase 3, decreased the expression of AKT.

CONCLUSIONThe effects of curcumin on inhibiting proliferation and promoting apoptosis of EC109 cells were related with increased expression of PTEN and inhibition of PI3K/AKT signaling pathway.

Apoptosis ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; Curcumin ; pharmacology ; Esophageal Neoplasms ; metabolism ; Glycogen Synthase Kinase 3 ; metabolism ; Glycogen Synthase Kinase 3 beta ; Humans ; PTEN Phosphohydrolase ; metabolism ; Phosphatidylinositol 3-Kinases ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Signal Transduction

AIM: To investigate the clinical effect for treating bullous keratopathy ( BK ) by anterior corneal stromal acupuncture combined with amniotic membrane transplantation.
METHODS: Totally 35 patients ( 35 eyes ) with bullous keratopathy were treated by corneal stromal acupuncture combined with amniotic membrane transplantation. All patients preoperative and postoperative underwent anterior segment OCT, corneal topography, corneal sensitivity and confocal microscopy. To observe postoperative ocular symptoms in patients with BK, recurrence of bulla, changes in corneal thickness, corneal sensitivity and changes in the organizational structure of the layers of the cornea.
RESULTS: Thirty-five were followed up for 6-18mo. The symptom of pain disappeared in 32 cases ( 91%) in the first day after operation and did not recur during follow-up. The symptom of pain relieved in 3 cases (9%) in the first day after operation and disappeared in 3 days. Corneal epithelium of 9 cases ( 26%) were all healed within 1wk, 21 cases (60%) were all healed within 2wk, and 5 cases (14%) were all healed within 3wk. Following up for 6-18mo, there was no recurrence of symptoms or bulla. A small amount of tiny bubbles in the surrounding area appeared in 2 cases after 3 and 4wk. All patients had no neovascularization, and had smooth corneal surface. The amnions of 30 cases (86%) were thinning after 2mo, partially dissolved and absorped, not seen with the naked eye after 3mo. Thirty-four cases (97%) had no changes in vision, one case (3%) was from the light to front of the manual. After 2mo, corneal sensation decreased in 30 cases ( 86%) , corneal thickness increased from preoperative 788±35μm to 940±43μm. After 12mo, corneal thickness increased to 1060±27μm. Results of confocal microscopy: after 3mo, the number density of the trigeminal nerve fibers under corneal basement membrane reduced, shallow stromal cells became into fibrotic stroma, deep stroma was more loose, and cells swelled significantly. The number of endothelial cells reduced and form swelled more obviously compared with preoperatively.
CONCLUSION: Corneal acupuncture combined with amniotic membrane transplantation can effectively control the symptoms of BK, prevent the recurrence of BK, and especially it is a simple, safe and practical way for patients with poor visual function.

Objective: To identify the functional domain of ?-Synuclein in affecting mitochondrial function and how the function to be impaired,especially,the mitochondrial membrane potential and the release of Cytochrome c.Methods: Harvest of ?-Syn-N and ?-Syn-△N by PCR,then subcloned into the pCMV-Myc mammalian expression vector.The recombinant plasmids were transfected into HEK293T cells by Lipofectamine 2000.After detecting the protein expression by Western blot,the functional domain was detected by co-immunoprecipitation.The mitochondrial membrane potential through flow cytometry and immunofluorescence,at the same time,the release of Cytochrome c through flow cytometry to detect.Results: The recombinant plasmids were constructed successfully.CO-IP has proved that N-terminal may be the functional domain of ?-Synuclein in affecting mitochondria.Over-expression of N-terminal could depolarize the mitochondrial membrane potential and induce the Cytochrome c releasing in MN9D cells.Conclusion: N-terminal may be the functional domain of ?-synuclein and over-expression of N-terminal could decrease mitochondrial activity.

OBJECTIVETo investigate the effect of Xianxiong decoction on the mice with acute lung injury induced by lipopolysaccharide.

METHODEighty female ICR mice were randomly divided into 8 groups: model group, Xianxiong decoction group, Daxianxiong decoction group, Xianxiong decoction group without Kansui Radix group, Xianxiong decoction group without Glycyrrhizae Radix et Rhizoma group, Glycyrrhizae Radix et Rhizoma and Kansui Radix group, normal group and control group. Animals of each group, except normal group, were undertaken intraperitoneal injection and intranasal inhalation of lipopolysaccharide (LPS) on day 1, 2, 3 to establish acute lung injury (ALI) model. 30 min after modeling, 0.2 mL corresponding drugs were administrated to each mice, dexam ethasone and normal saline were given to the mice of control group and normal group respectively. White blood cell in blood, neutrophil percentage of blood and bronchoalveolar lavage fluid (BALF) supernatant, the ratio of wet and dry lung tissue ( W/D), histopathological changes of lung tissue were estimated. Sixty ICR mice were randomly divided into normal, model, control, high, middle and low dose Xianxiong decoction groups and were modeled in the same way. ELISA was applied to detect the level of NF-kappaB, TNF-alpha and IL-6 in BALF, PCR for NF-kappaB and TNF-alpha mRNA in lung tissue, and Western blot for NF-kappaB and TNF-alpha. Half of 20 ICR mice were administrated with Xianxiong decoction of its maximum tolerant normal saline.

RESULTCompared with model group, the number of WBC in blood of Xianxiong decoction group mice decreased (P < 0.01), percentage of neutrophils in both blood and BALF decreased as well (P < 0.01, P < 0.05); it also significantly reduced the ratio of W/D (P < 0.01); and found the alveolar wall, the number of inflammatory cells infiltrating improved, compared with model group. Xianxiong decoction reduced the level of NF-kappaB, TNF-alpha and IL-6 in BALF (P < 0.01, P < 0.01, P < 0.05); its high and low dose groups only found TNF-alpha level declined. Five mice died 24 h after administration of Xianxiong decoction which indicated its toxicity when other influential factors were considered.

CONCLUSIONXianxiong decoction is effective on the ALI mice induced by LPS, but it is of toxicity at 3 g x mL(-1).

Acute Lung Injury ; drug therapy ; genetics ; metabolism ; pathology ; Animals ; Bronchoalveolar Lavage Fluid ; chemistry ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Interleukin-6 ; genetics ; metabolism ; Lipopolysaccharides ; adverse effects ; Lung ; drug effects ; metabolism ; pathology ; Mice ; Mice, Inbred ICR ; NF-kappa B ; genetics ; metabolism ; Tumor Necrosis Factor-alpha ; genetics ; metabolism

AIM: To explore the characteristic of relaxation of porcine cornea after LASIKMETHODS: Usual LASIK was performed on fresh porcine corneas with stable intraoculer pressure (IOP) maintained through optic nerve irrigation. The ablation depth on stroma is 30%, 50% and 70% respectively. Then the dumbbell-shaped corneal strip specimens were cut and stored in 20% Human Albumin solution for use (4℃).Sip up the Albumin solution on specimens, and fixed them on homemade jig. Stress relaxation tests were erformed on Tytron250 Dynamics Experiment System. the loading speed was 385mm/min,extending ratewas 1.5, and relaxation time was 1 000s. The data werecollected electronically and automatically.RESULTS: In LASIK procedure, though a single flap-cutting can cause a little reduction of corneal stress relaxation (P＜0.05,P=0.49), the cornea may still remain its property of visco-elasticity. When ablation depth was 30% or more, corneal stress relaxation decreased to almost one half (P＜0.01).The change of corneal stress relaxation degree in vertical meridian specimen was lower than that in horizontal specimen, especially when ablation depth was 70%, and it's statistically significant (P＜0.001). In LASIK operation, the more depth the ablation, the more reduce the stress relaxation degree, and it's easy to cause deformation and creep deformation.CONCLUSION: The changes of the stress relaxation in verticai and longitudinal meridian specimens are similar, and slightly obvious in longitudinal specimen, especially in 70%ablation group.

Objective To investigate the clinical significance of D-dimer in systemic lupus erythematosus (SLE). Methods The study group comprised 261 SLE patients who were admitted in ward from 2005 to 2008 in Peking University People's Hospital Collect the clinical data to investigate the clinical significance of D-dimer. Results ( 1 ) The D-dimer levels of 56 patients were increased due to coexist reduced renal function, infections, disseminated intravascular coagulation ( DIC), liver disorders, pregnancy and injury. With the exception of above patients, 142 ( 69. 3 ) patients were increased in total 205 patients. (2)The level of D-dimer was positively correlated with SLE disease activity index( SLEDAI ) score ( r =0. 598,P =0. 000), and was associated with anti-dsDNA antibody, ESR, C-reactive protein(CRP) and complement C3 and C4. (3)D-dimer level was associated with important organ involvement. (4)All patients with thrombosis had increased D-dimer, but patients without thrombosis had normal or increased D-dimer levels. Conclusion The level of D-dimer elevates in patients with active disease or important organ involvement , it can not identify thrombosis. All patients with thrombosis had increased D-dimer levels.

Abstract: Objective To analyze the effect of nutrition package on the nutritional status and prevalence of children in rural areas of Hainan Province, and provide scientific basis and suggestions for further improving the nutritional and health status of children in this region. Methods　Four cities and counties were randomly selected as the intervention group, and four cities and counties matched with the intervention group in terms of population, economy, social culture, maternal and child health work foundation of township health centers, physical nutrition and health status were selected as the control group.With the combination of monitoring and prospective cohort study, infants in the intervention group and the control group were studied from June 1, 2020, and they were intervened for 12 months with supplementary food nutrition package. Before and after intervention, the nutrition and health status of infants aged 6-24 months in the intervention group and the control group were investigated to evaluate the nutritional and health effects of supplementary food nutrition package for infants aged 6-24 months in rural Hainan Province. Results　A total of 999 infants were investigated, including 427 in the intervention group and 572 in the control group. After 12 months of nutritional intervention, there was no significant difference in weight-for-age Z-score (WAZ) and height-for-age Z-score (HAZ) and weight-for-height Z-score (WHZ) between the intervention group and the control group (P>0.05). The rate of emaciation of the intervention group was 1.64%, which was significantly lower than 3.67% of the control group (P<0.05). There were no significant differences in the rate of growth retardation (2.81% and 3.32%, respectively) and underweight (0.47% and 1.92%, respectively) between the intervention group and the control group (P>0.05). The rate of respiratory infection and diarrhea in the intervention group were 9.13% and 1.17%, which were significantly lower than corresponding 23.25% and 3.15% in the control group (P<0.05). The hemoglobin of the intervention group and the control group were 117.24 g/L and 114.51 g/L respectively, and the rates of anemia were 11.11% and 22.84% respectively, the differences were statistically significant (P<0.05). Conclusions　The intervention of nutrition package in rural areas of Hainan Province has achieved the expected results, and supplementary food nutrition package has reduced the incidence of malnutrition and respiratory infection and diarrhea in recent two weeks in infants and anemia to a certain extent. We should attach great importance to the supplementary nutrition package for right-age children and promote the growth and health of children in rural areas through supplementary nutrition package, and continuously improve the nutrition and health level of children in Hainan Province.