AIM: To explore the feasibility of culturing human umbilical vein endothelial cells ( HUVEC ) on acellular corneal stroma and performing the posterior lamellar endothelial keratoplasty ( PLEK ) treating corneal endothelial decompensation.
METHODS: Thirty New-Zealand rabbits were divided into three groups randomly, 10 rabbits for experimental group, 10 for stroma group and 10 for control group. Corneal endothelial cells were removed to establish animal model of corneal endothelial failure. PLEK was performed on the rabbits of experimental group and stroma group, and nothing was transplantated onto the rabbits of control group with the deep layer excised only. Postoperative observation was taken for 3mo. The degree of corneal edema and central corneal thickness were recorded for statistical analysis.
RESULTS:Corneas in experimental group were relieved in edema obviously compared with that in stroma group and the control group, and showed increased transparency 7d after the operation. The average density of endothelial cells was 2 026. 4±129. 3cells/mm2 , and average central corneal thickness was 505. 2±25. 4μm in experimental group, while 1 535. 6±114. 5μm in stroma group and 1 493. 5±70. 2μm in control group 3mo after operation.
CONCLUSION:We achieved preliminary success in our study that culturing HUVEC on acellular corneal stroma and performing PLEK for corneal endothelial decompensation. HUVEC transplanted could survive in vivo, and have normal biological function of keeping cornea transparent. This study provides a new idea and a new way clinically for the treatment of corneal endothelial diseases.

Objective To investigate the characteristics and risk factors of abdominal lymph node metastasis in thoracic esophageal squamous cell cancer.Methods The clinical data of 586 patients with thoracic esophageal cancer who underwent surgery via transabdominal and transthoracic approaches between June 2009 and June 2014 at the Sichuan Cancer Hospital were retrospectively analyzed.All the patients received resection of esophageal cancer and lymph node dissection, and the transabdominal right thoracic approach or cervico-thoracicabdominal triple incision was selected according to the condition of patients.No.18, 19, 20 lymph nodes were dissected seperately and No.16, 17 and lesser curvature lymph nodes were separated.All the specimens of lymph nodes were detected by regular pathological examination.Measurement data with normal distribution were presented as x ± s and count data were described as rate.Comparisons of rate between 2 specimens and among the multiple specimens were respectively analyzed using the chi-square test and partition of chi-squared.The multivariate analysis was done using the logistic regression.Results The number of lymph node dissected in 586 patients was 12 524 with an average number of 20 ± 11 per case, and the rate of lymph node metastasis was 55.63％ (326/586).The number of mediastinal lymph node dissected was 7 012 with an average number of 12 ± 5 per case, and a rate of mediastinal lymph node metastasis was 40.96％ (240/586).The number of abdominal lymph node dissected was 5 512 with an average number of 9 ± 8 per case, and a metastasis rate was 31.74％ (186/586).The abdominal lymph node metastasis rate of the upper, middle and lower thoracic esophageal cancer were 13.73％ (14/102), 31.51％ (92/292) and 41.67％ (80/192), respectively, showing a significant difference among the above 3 indexes (x2 =25.91, P ＜ 0.05).The lymph node metastasis rate in No.16, 17, 18,19, 20 and lesser curvature lymph nodes were 12.80％ (75/586), 16.89％ (99/586), 1.71％ (10/586),0.68％ (4/586), 1.71％ (10/586) and 2.05％ (12/586), respectively, with a significant difference among the above 6 indexes (x2 =287.95, P ＜ 0.05).The results of univariate analysis showed that the tumor location,surgical procedure, T stage, N stage, G stage, pathological stage and mediastinal lymph node metastasis were risk factors affecting abdominal lymph node metastasis of thoracic esophageal cancer (x2 =24.02, 23.97, 37.87,136.85, 38.79, 7.70, 154.27, P ＜ 0.05).The tumor in the lower thoracic portion, N3 stage and stage Ⅳ were independent risk factors affecting abdominal lymph node metastasis of thoracic esophageal cancer in the multivariate analysis (RR =5.80, 2.36, 2.76, 95％ confidence interval: 1.022-1.813, 1.317-3.950, 1.652-12.351, P ＜ 0.05).Conclusions Abdominal lymph node metastasis is common in thoracic esophageal cancer in which No.16 and 17 lymph nodes predominate, and it is easy to occur in patients with lower thoracic esophageal cancer, and advanced N stage and pathological type.

Objective: To observe the difference in anti-inflammatory effect between living rhino horn and rhino horn by the method of comparative research to provide the experimental basis for the replacement of rhino horn by living rhino horn.Methods: The anti-inflammatory effects of living rhino horn and rhino horn were studied by the methods of paw edema in rats, cotton ball granuloma in mice, auricle swelling and peritoneal dye penetration.Results: Compared with that in the model control group, the foot metatarsus swelling degree at all time points in high (440 mg·kg-1) dose group and middle (220 mg·kg-1) dose group of living rhino horn and three doses groups of rhino horn showed statistical differences (P＜0.05 or P＜0.01).The high dose group (700 mg·kg-1) and middle dose group (350 mg·kg-1) of living rhino horn and rhino horn could significantly reduce the weight of cotton ball granuloma in mice (P＜0.05).Three doses groups (700, 350 and 175 mg·kg-1) of living rhino horn and rhino horn could significantly reduce auricle swelling in mice induced by xylene (P＜0.05 or P＜0.01).The absorbance of Evansan in the abdominal cavity in the middle dose group (350 mg·kg-1) of rhino horn and the high dose group (700 mg·kg-1) and middle (350 mg·kg-1) dose group of living rhino horn was significantly reduced (P＜0.05 or P＜0.01).There was no significant difference in the anti-inflammatory effect between living rhino horn and rhino horn at the same dose.Conclusion: Living rhino horn and rhino horn have a certain anti-inflammatory effect.The anti-inflammatory effect of living rhino horn is similar to those of the rhino horn, and living rhino horn can be used as a substitute of rhino horn.

The bleeding in liver resection is an important factor influencing the operation risk and postoperative out comes.Hepatic vascular exclusion technique is an effective method to control the bleeding,which has been improved after being widely used in clinical practice.In this paper,the research progress on the clinical application of various kinds of hepatic vascular exclusion techniques was reviewed.

OBJECTIVETo observe the effect of β₃adrenoceptors (β₃-AR) activation on rat thoracic aorta smooth muscle contractility and the possible related mechanism.

METHODSThe endothelium removed thoracic aorta was pre-contracted with 30 mmol/L KCl physiological saline solution (PSS). Then the tension of the thoracic aorta was recorded in presence of BRL37344 (BRL) to determine the action of β₃-AR. The tension of the thoracic aorta was also recorded in the presence of Propranolol (PRA), SR59230A (SR), L-NNA, H-89 and Iberiotoxin (IBTX) respectively to reveal the underling mechanism of β₃-AR activation on rat vascular smooth muscle. Immunohistochemistry was adopted to confirm the existence and the distribution of β₃-AR in rat thoracic aorta.

RESULTSThe results showed that: (1) The thoracic aorta was relaxed by β₃-AR activation, with a relaxation percentage of (10.59 ± 0.79). (2) β₃-AR was expressed in both endothelial and smooth muscle layer in thoracic aorta sections of rats. (3) PRA did not block the effect of BRL on the thoracic aorta. The relaxation actions of BRL could be antagonized by pre-incubating the thoracic aorta with SR. (4) L-NNA (a NOS inhibitor) and H-89 (a PKA inhibitor) reversed the relaxation effect of BRL on vascular smooth muscle. (5) The effect of BRL was decreased after application of Ibriotoxin (IBTX), a large conductance calcium dependent potassium channel blocker.

CONCLUSIONThe results confirmed that activation of β₃-AR led to relaxation of thoracic aorta smooth muscle. The relaxation action of β₃-AR on smooth muscle of rat thoracic aorta was related to activation of NOS and PKA signaling pathway. Large conductance Ca²⁺-K⁺ channels were involved in the relaxation action of β₃-AR activation on rat thoracic aorta smooth muscle.

Animals ; Aorta, Thoracic ; physiology ; In Vitro Techniques ; Isoquinolines ; Large-Conductance Calcium-Activated Potassium Channels ; physiology ; Muscle Contraction ; Muscle Relaxation ; Muscle, Smooth, Vascular ; physiology ; Nitroarginine ; Peptides ; Propanolamines ; Propranolol ; Rats ; Receptors, Adrenergic, beta-3 ; physiology ; Signal Transduction ; Sulfonamides

Objective To establish reference values for blood pressure in cynomolgus monkeys in different ages.Methods The blood pressures and blood lipids indexes were detected in 521 cynomolgus monkeys using an American BECKMAN-CX4 automatic biochemical analyzer and a wrist electronic blood pressure monitor.Statistical tests were performed to analyze the data.Results Significant differences were found in blood pressure values of cynomolgus monkeys in different ages.Blood pressure values in the elderly group were higher than those of other groups.The morbidity of hypertension in the elderly group was higher than those of the other groups.Body mass index (BMI) in the hypertension group was higher than that of normal group in the same age.The incidence of hypertension in the elderly group with hyperlipemia was higher than that of other groups.Logistic regression analysis showed that age, BMI and hyperlipidemia in the hypertensive group were 1.435, 1.218, and 2.337 times higher than those of the normal group when predicting the risk of hypertension.Conclusions We have initially established reference values of blood pressure in cynomolgus monkeys in different ages.Age, BMI and hyperlipidemia are risk factors of spontaneous hypertension in cynomolgus monkeys, and the measurement of blood pressure may provide a basis for the screening of cynomolgus monkey model of spontaneous hypertensive and related research.

Objective To investigate the effect of preconditioning with different concentrations of sevoflurane on hypoxia-reoxygenation(H/R)-induced apoptosis in rat hippocampal neurons and the role of mitochondrial KATP(mito-KATP)channels.Methods Primary cultured hippocampal neurons isolated from newborn SD rats(＜24h)of both sexes,weighing 5-6 g,were randomly divided into 7 groups with 48 wells and 12 dishes in each one:control group(C group),H/R group,preconditioning with 6%,4%and 2% sevoflurane groups(S1-3 groups),5-hydroxydecanoate(5-HD,mito-KATP channel blocker)100 μmol/L preconditioning group(5-HD group)and preconditioning with 5-HD 100 μmol/L+6% sevoflurane group(5-HD+S group).The neurons were exposed to 4 h hypoxia followed by 24 h reoxygenation. In S1-3 groups, preconditioning was performed with 6% , 4% and 2% sevoflurane respectively before H/R. In 5-HD group, preconditioning was performed with 5-HD (final concentration 100 μmol/L) before H/R. In 5-HD + S group, preconditioning was performed with 5-HD 100 μmol/L and 6% sevoflurane before H/R. The neuronal viability, apoptosis rate and expression of Bcl-2 and Bax were determined after 24 h reoxygenation.Results The neuronal viability was significantly lower,while the apoptosis rate and expression of Bcl-2 and Bax were significantly higher in the other 6 groups than in group C(P＜0.01).The neuronal viability and expression of Bcl-2 were significantly higher,while the apoptosis rate and Bax expression were lower in S1-3 groups than in group H/R. There was no significant difference in the parameters mentioned above between 5-HD and 5-HD + S groups(P＞0.05).The neuronal viability and expression of Bcl-2 were significantly lower, while the apoptosis rate and Bax expression were higher in S2, S3 and 5-HD + S groups than in group S1, and in group S3 than in group S2(P＜0.0l) .Conclusion Sevoflurane preconditioning can inhibit H/R-induced apoptosis in rat hippocampal neurons and reduce the injury to neurons in a concentration-dependent manner, and the underlying mechanism may be related to activation of mito-KATP channels, up-regulation of Bcl-2 expression and down-regulation of Bax expression.

A new on-column conductivity detection for microchip capillary based on the accessional solution conductor between separation channel and electrodes was designed. In this detection, the carrier electrolyte solution was chosen as conductor to eliminate large drifts of the detection signals, that because different composition or concentration can lead to transport processes(diffusion and electromigration) between the carrier electrolyte solution and mediator. The separation channel incorporated a dual-T and a crux-channel, dual-T was used for sample introduction, and the crux-channel was used for detection. The sensing electrodes connected to separation channel through the crux-channel and obtained the potential different of conductivity. This method avoids integrating the sensing electrodes directly within the separation channel and prevents any direct contact of the electrodes with the sample. It not only avoids electrode reactions but also is convenient for washing and replacing electrodes. In addition, the signals, contrary to contactless conductivity detection, can be detected in the low excitation voltage(2.5-4.0 V) and frequency(700-1700 Hz) range. In the buffer solution of 15 mmol/L MES-His(pH 5.8), the detection limits was 0.5 and 0.1 μmol/L for K+ and Na+.

A new teaching method was developed in the curriculum of Harmful Microorganisms Control Technology.It is characterized by students’ self-learning followed by student’s instruction.Both students and teacher have succeeded in this model after four stages of practice,in which a pleasant learning atmosphere was created in the classroom.An effective interaction between teacher and students was achieved.Students are viewed as main objects in the classroom and they are encouraged to ask questions,to formulate their own ideas,or to find things out for themselves.Thus,students’ abilities including presentation,communication,competition,and cooperation were enhanced.By adapting their role to the new teaching method,teachers have also improved their teaching skill and strategies.