Objective To assess the effect of cryopreservation of cornea in glycerol,to analyze the protective effect of sodium hyaluronate on corneal endothelial cells(CEC) during preservation,and to find out a convenient and cheap method for the storage of cornea.Methods 20 rabbit corneas were divided into two groups randomly.The corneas of control group were stored directly in pure glycerol at-25℃,while in the experimental group sodium hyaluronate was evenly anointed on the corneal endothelial surface of the cornea before being cryopreserved in glycerol.2 months later,the effect of preservation for both groups was examined with trypan blue combined with alizarin red staining(viability staining) for corneal endothelial cells,electron-microscope examination for preserved corneas,and rabbit experimental penetrating keratoplasty(PKP) with the long-term cryopreserved corneas from both the experimental and control groups.Results CEC viability could be preserved by both methods.CEC trypan blue combined with alizarin red staining(viability staining) showed that the CEC density of cornea in the experimental group was 3 067.9?127.6/mm2,which was higher than that of control group(2 899.9?134.0/mm2).The mortality of CEC in experimental group was 4.2%?3.4%,lower than that in control group(13.8%?2.2%).Suceessful rate of PKP was higher in the experimental group with higher CEC density.Conclusion Cryopreservation with glycerol can preserve the viability of CEC.Sodium hyaluronate can reduce the damage to endothelium in the preserving procedure and can obviously improve the effect of preservation.The improved cryopreservation of cornea with glycerol may be a convenient,economic and effective method for eyebank.

ObjectiveTo detect the mechanism of the growth inhibition and apoptosis of human acute leukemia cell line U937 cells induced by honokiol.MethedsThe proliferation inhibition was detected by MTT method.Cell apoptosis was tested by Hoechst 33342 staining and flow cytometry with Annexin V/PI staining.RT-PCR was used to detect the mRNA expression of the apoptosis gene Bcl-2,Bax,Caspase 3,Caspase 8 and Caspase 9.ResultsThe inhibition effect of honokiol(5 μg/ml,48 h) on U937 cells proliferation could he observed,and the inhibition rate of 10 μg/ml honokiol on cell proliferation reached above 50％ (48 h).U937 cells proliferation could be completely inhibited for 120 h. U937 cells apoptosis rate reached 26.8％ (P ＜0.01)after being treated with 10 μg/ml honokiol.After being treated with 10 μg/ml honokiol for 48 h,the Bcl-2 gene expression in U937 cells was reduced (control group:0.33 ± 0.02,experimental group:0.14 ±0.01,P ＜ 0.01 ),and the Bax gene expression was elevated ( control group:0.1 ± 0.01,experimental group:0.87 ± 0.08,P ＜ 0.01 ).The gene expressions of Caspase 3 ( control group:0.48 ± 0.01,experimental group:0.87±0.06,P ＜0.01),Caspase 8(control group:0.23±0.02,experimental group:0.41 ±0.07,P ＜ 0.01 ) and Caspase 9 ( control group:0.44 ± 0.05,experimental group:0.76 ± 0.06,P ＜ 0.01 ) were all increased.The activity of Caspase-3 was 0.325 ±0.089,which was significantly higher than that of the control group ( P ＜0.01 ).ConclusionHonokiol can significantly inhibit the proliferation and induce cell apoptosis of human acute leukemia cell line U937 cells.The mechanism is related to the up-regulation of Bax and down-regulation of Bcl-2,and the endogenous and exogenous pathways are both inolved in the apoptosis process.

Objective To establish an effective post-operative oral care procedure for patients with oral cavity cancer.Methods Eighty five post-operational patients with oral cavity cancer admitted to Sichuan Cancer Hospital during March 2011 and March 2013 were randomly divided into control group (42cases) and study group (43 cases).Patients in control group were given routine oral care,and those in study group received a special care procedure:normal saline washing + dioxogen cleaning + normal saline washing + normal saline gargle.Oral cleanliness,oral and lung infections were observed in both groups,and SPSS 17.0 was used for statistical analysis.Results Oral cleanliness,rates of oral infection and lung infection at d3 after operation,time of infections,the delayed wound healing and operating comfort were improved in the study group (Z/t/x2 =-2.48,4.57,4.64,2.36,2.25,-2.51,4.18,respectively; all P ＜ 0.05).Conclusion The oral care procedure containing normal saline washing + dioxogen cleaning +normal saline washing + normal saline gargle can improve oral cleanliness early after operation,reduce complications and increase the operating comfort.

Objective To investigate the value of intraoperative neuromonitoring(IONM) for preventing recurrent laryngeal nerve(RLN) injury in thyroid surgery.Methods 1087 consecutive patients undergoing thyroid surgery at Wuhan Central Hospital from Oct 2010 to Dec 2012 were evaluated.277 cases(the study group) underwent thyroidectomy with RLN identification by naked eye plus IONM.810 cases(the control group)underwent thyroidectomy with RLN identification by naked eye only.Results In the control group,733 RLNs (90.49％,733/810)were successfully identified and 77 RLNs were failed to be identified.In the study group,all the 277 RLNs(100％,277/277)were successfully identified.28 cases had postoperative temporal RLN injury,among whom 26 cases were in the control group and 2 cases were in the study group.18 cases in the control group and 2 cases in the study group recovered in 2 weeks after surgery.The rest recovered in 2 months after surgery.RLN injury rate was significantly lower in the study group than in the control group.There was no significant difference in RLN injury for low-risk surgical patients between the 2 groups; however,the study group had advantages in RLN injury than the control group for high-risk patients.Conclusions IONM in thyroid surgery can improve the recognition rate of RLN during thyroidectomy.IONM can significantly reduce the incidence of RLN injury,especially in high-risk surgery.

It is necessary to medical student to take the elective course of pharmacy.This paper put forward several viewpoints and concrete methods about teaching methods,means and contents of elective course of pharmacy.

Objective To investigate 13 high-risk types of HPV (HR HPV) infection rates in women with different grades of cervical lesions.Methods A total of 350 women, who were hospitalized in the department of gynecology in Bao′an Maternity & Child health hospital, were enrolled for the study.TCT technology was used to evaluate the cervical epithelium.The group were divided according to the cytology results.Multiplex real time PCR (mRT PCR) was used to detect the viral loads.HR HPV infection rate of different groups were analyzed using χ2 test or Fisher exact test.HR HPV viral loads of patients in different grades of cervical lesion groups were compared using Kruskal-Wallis or Wilcoxon test, and the age distribution of HR HPV positive group and negative group was analyzed by using Wilcoxon test.Results The HR HPV infection rates of NILM, ASCUS, LSIL, HSIL were 3.4% (10/295), 20.0% (7/35), 78.6% (11/14) and 100.0% (6/6), respectively.HR HPV positivity in NILM was lower than ASCUS (χ2=14.43,P<0.01) and LSIL (χ2=107.69,P<0.01), HR HPV positivity in ASCUS was lower than LSIL (χ2=14.76,P<0.01). The median of HR HPV viral loads in NILM, ASCUS, LSIL and HSIL were 4.10 (3.38-6.27), 5.33 (3.63-6.66), 5.77 (4.01-7.01) and 5.58 (4.19-5.85) respectively (copies/ml,lg).Combined ASCUS, LSIL and HSIL groups into cervical lesion group, HR HPV viral load of which was higher than that of NILM (U=43.0, P<0.05).The median Ages of HR HPV positive group and negative group were 36 and 33, respectively.No statistical significance was found between them (U=4 544, P>0.05).Conclusions The present study revealed that HR HPV infection was related to cervical lesion, but there was no correlation between viral load and cervical lesion grade. In additional, no difference in age distribution was found between HR HPV positive group and negative group.

An ancient genome duplication (PPP1) that predates divergence of the cereals has recently been recognized. We report here another potentially older large-scale duplication (PPP2) event that predates monocot-dicot divergence in the genome of rice (Oryza sativa L.), as inferred from the age distribution of pairs of duplicate genes based on recent genome data for rice. Our results suggest that paleopolyploidy was widespread and played an important role in the evolution of rice.
Biological Evolution ; Chromosome Mapping ; methods ; Evolution, Molecular ; Genetic Variation ; genetics ; Genome, Plant ; Oryza ; genetics ; Polyploidy

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

Objective To compare the effects of airway humidification by intermittent atomization inhalation and micro pump control on pulmonary infection after tracheotomy in elderly patients with serebral hemorrhage. Methods From February 2012 to February 2013, 46 elderly patients with heavy craniocerebral injury and tracheotomy were set as the control group, treated with intermittent atomization inhalation. From March 2013 to March 2014, 50 elderly patients with severe craniocerebral injury and tracheotomy were set as the observation group, treated with micro pump controlled continuous airway humidification. The two groups were compared in terms of the satisfaction rate and incidence of pulmonary infection. Result The satisfaction rate of the observation group was higher than that of the control group and the incidence of pulmonary infection was significantly lower than that of the control group (P<0.05). Conclusion Micro pump controlled airway humidification for elderly patients with severe craniocerebral injury can improve the airway humidification effect and reduce the incidence of pulmonary infection.

ObjectiveTo study the effect of pre-arrest and post-arrest mild hypothermia after restoration of spontaneous circulation (ROSC) on myocardial function, ultrastructure, apoptosis of myocardial cells in rabbits with ventricular fibrillation.Methods Sixty-two male New Zealand rabbits were randomly allocated into five groups: namely normothermic control group (NTC group,n = 10), hypothermia control group (HTC group,n = 10), normothermic resuscitation group (NTR group,n = 14), hypothermia pre-arrest group (HPRA group,n = 14), and hypothermia post-arrest group (HPOA group,n = 14). The normal temperature was controlled at (39.0±0.5)℃, and the hypothermia (33.5±0.5)℃. Ventricular fibrillation cardiac arrest (CA) was reproduced in rabbits by transcutaneous epicardium electrical stimulation. The parameters of hemodynamics were monitored dynamically for 4 hours in all the groups, including heart rate (HR), left ventricular end diastolic and systolic pressure (LVEDP/LVESP), maximal rate of increase/decrease in left ventricular pressure (±dp/dt max), and mean arterial pressure (MAP). The body temperature of rabbits in hypothermia groups was maintained by surface cooling for 4 hours followed by rewarming. The survived rabbits were sacrificed at 48 hours after resuscitation, and myocardial apical tissue was harvested for observation of ultrastructure with electronic microscope, and to observe apoptosis by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining.Results① Resuscitation investigation: there was no significant difference in rate of ROSC, time of CPR and energy of defibrillation among HPRA, HPOA, and NTR groups [rate of ROSC: 85.71%, 71.43%, 71.43%; time of CPR (seconds): 45.3±30.2, 61.2±41.3, 82.3±63.8;energy of defibrillation (J): 14.3±8.9, 22.0±15.5, 25.0±15.8, allP> 0.05].② Hemodynamics: compared with normal temperature groups, animals in hypothermia groups exhibited lower levels of HR (allP< 0.05). Compared with NTR group, HPRA group exhibited higher levels of LVESP (mmHg, 1 mmHg = 0.133 kPa) at 0.5, 1, 2 and 3 hours post ROSC (0.5 hour: 103.8±14.3 vs. 91.6±13.3, 1 hour: 107.2±14.1 vs. 82.7±8.5, 2 hours: 109.0±16.9 vs. 88.8±12.9, 3 hours: 109.1±14.6 vs. 89.3±14.3, allP< 0.05). Compared with NTR group and HPOA group, HPRA group exhibited lower levels of LVEDP (mmHg) at 0.5 hour post ROSC (3.70±0.85 vs. 7.61±2.73, 7.02±3.12, both P< 0.05). Compared with NTR group, HPRA group exhibited lower levels of LVEDP at 1 hour post ROSC (4.34±1.44 vs. 6.99±1.96,P< 0.05). In HPRA group, the level of+dp/dt max (mmHg/s) was higher than that of NTR group and HPOA group at 1 hour and 2 hours post ROSC (1 hour: 2 759.5±321.6 vs. 2 123.0±304.5, 2 283.7±234.2, 2 hours:2 730.6±425.1 vs. 2 221.5±392.9, 2 252.6±476.0, allP< 0.05). There were no significant differences in -dp/dt max and MAP levels among three CPR groups.③ The survival rate at 48 hours post ROSC of NTR, HPRA and HPOA groups was 60%, 75%, and 100%, respectively. Compared with NTR group, higher survival rate was found in HPOA group at 48 hour post ROSC (P< 0.05).④ Compared with NTR group, less damage to myocardial ultrastructure was found in HPRA and HPOA groups. Apoptosis index (AI) was lower in HPRA and HPOA groups than that in NTR group [(28.05±9.82) %, (26.39±8.98) % vs. (42.02±13.36) %, bothP< 0.05].Conclusions Our study shows that mild hypothermia has no effect on ROSC rate. Pre-arrest hypothermia can ameliorate myocardial systolic function of rabbit in early stage after ROSC, and it has no negative influence on diastolic function. Post-arrest mild hypothermia produces no negative influence on myocardial function of rabbit, but it improves 48 hours survival rate in ROSC rabbits. Both pre-arrest and post-arrest mild hypothermia therapy can attenuate myocardial injury in CA model of rabbits by ameliorating mitochondrial injuries and suppressing apoptosis of myocardial cells.