AIM: In order to search carrier material and better tissue culture method, the morphology and structure of the cultured cat corneal endothelium was observed. METHODS: The cat corneal endothelial cells were seeded on the Descemet's membrane of the dehydrated swine corneal stroma, and then cultured in the medium with epidermal growth factor and laminin for 7 days. The morphology and structure of reconstructed tissue was tested by the inverted microscope and the scanning electron microscope. RESULTS: As shown by the morphological observations, the cultured cat corneal endothelial cells formed an integrated membrane, and attached to the Descemet's membrane, which was similar to the nature tissue. The cells connected tightly to each other, and some of them arranged in hexagon approximately. CONCLUSION: The cat corneal endothelial cells could be rebuilt on the carrier of the dehydrated swine corneal stroma successfully, which is similar to the nature tissue.

0.05). (2) In the total 12 rabbits, all the cornea grafts survived without rejection reaction, corneal haze or corneal neovascularization. The fresh grafts got transparent after 2 months, and the dehydrated grafts got transparent after 6 months. Histological study demonstrated both fresh and dehydrated stroma grafts had fused with rabbits'corneal stroma very well without lymphocytes infiltrating. (3) As shown in histological observations, the reconstructed cat corneal endothelial tissue was similar to the nature tissue. Cultured cat endothelial cells connected tightly to each other and attached to the Descemet's membrane closely. CONCLUSION: Swine corneal stroma has low immunogenicity and satisfying biocompatibility,it is an ideal biological carrier for cornea reconstruction in vitro.

Objective To discuss the perioperative nursing care for patients with hepatic cancer located near the central bile duct, who receive microwave ablation treatment by using bile duct cooling protection technique. Methods A total of 21 patients with hepatic cancer located near the central bile duct received percutaneous microwave ablation treatment (PMWA) using bile duct cooling protection technique. Preoperative routine nursing and psychological nursing, close cooperation, close postoperative observation and prompt nursing measures were strictly carried out. Results PMWA was successfully accomplished in all the 21 patients, and no biliary complications occurred. All 21 patients were cured at the time of discharge, no nursing-related complications or death occurred. Conclusion In treating hepatic cancer located near the central bile duct, bile duct cooling protection technique is a new treatment technology. Effective perioperative nursing care can improve the operative success rate and reduce the incidence of complications.

OBJECTIVETo investigate the changes between pressure of trochlea of talus surface and distribution of area after anterior lower tibiofibular ligament rupture, and provide basis for treating anterior lower tibiofibular ligament rupture.

METHODSSix fresh adult ankle joint specimens (4 males and 2 females, ranging age from 25 to 60 years, with an average of 44.6 years) were adopted. The specimens were removed from skin and muscles, remained ankle joint capsule, medial and lateral ligaments and anteroinferior tibiofibular ligament. The ankle joint was fixed with a special fixture in neutral position. Pressure sensitive film (700 N axial load ) was respectively used to measure mean pressure, peak pressure and stress distribution area of the upper articular facet of talar trochlea of the normal ankle joint and the ankle joint with anterioinferior tibiofibular ligament rupture.

RESULTSThe stress distribution areas of the control group and the ruptured group were respectively (367.8 +/- 54.0) mm2 and (386.0 +/- 53.7) mm2; the mean pressures were respectively (1.40 +/- 0.12) MPa and (1.70 +/- 0.35) MPa; the peak pressures were respectively (2.60 +/- 0.33) MPa and (3.20 +/- 0.32) MPa. The experimental results showed that the change in stress distribution area after anterioinferior tibiofibular ligament rupture was not significant (t = 0.021, P = -0.983). When stress distribution changed, the region of stress concentration transferred to poster lateral,and mean pressure (t = 4.140, P = 0.020) and peak pressure (t = 3.169, P = 0.010) increased significantly.

CONCLUSIONWhen anterior lower tibiofibular ligament rupture occurs, mean pressure,peak pressure and stress distribution of pressure of trochlea of talus surface is changed, which may cause traumatic arthritis, and surgical treatment is considerably used to restore normal anatomy.

Adult ; Ankle Injuries ; Biomechanical Phenomena ; Female ; Fibula ; Humans ; Lateral Ligament, Ankle ; injuries ; Male ; Mechanical Phenomena ; Middle Aged ; Rupture ; Stress, Mechanical ; Tibia

Objective To explore the relationship between Arg913Gln(G→A) polymorphism of solute carrier family 12 member 3 (SLC12A3) gene and diabetic nephropathy (DN) in type 2 diabetes mellitus (T2DM) in Han population of Shanghai. Methods Two hundred and fifty-eight Han ethnic people in Shanghai with T2DM (T2DM group) were divided into non-DN group (DN0 group, n=95) and DN group (n=163) according to 24 h urine albumin excretion rate (AER), and those in DN group were subdivided into microalbuminuria group (DN1 group, n=95) and macroalbuminuria group (DN2 group, n=68). Besides, 82 people with normal results of oral glucose tolerance test (OGTT), without diabetes mellitus and nephropathy were served as controls. PCR-sequencing was used to detect the genotypes of Arg913Gln polymorphism of SLC12A3 gene. Genotypic and allelic frequencies and clinical characteristics were compared among groups. Results Three genotypes (GG, GA and AA) were detected. The frequencies of GA+AA genotype and A allele in T2DM group were higher than those in control group, while there was no significant difference between groups (P>0.05). There was no significant difference in genotypic or allelic frequencies among subgroups of T2DM group (P>0.05). The level of triglyeeride (TG), AER, level of fasting insulin (FINS) and HOMA-IR in patients with GA+AA genotype were significantly higher than those in patients with GG genotype in T2DM group (P<0.05). Conclusion Arg913Gln(G→A) polymorphism of SLC12A3 gene is not significantly associated with T2DM and DN in Han population of Shanghai. The AER of people with GA+AA genotype is significantly higher than that with GG genotype. Arg913Gln (G→A) polymorphism of SLC12A3 gene may predict the risk of increase of albuminuria in patients with T2DM in Han population of Shanghai.

ObjectiveTo apply the Schwann cell neurilemma channel(SCNC) in promoting the regeneration of injuried spinal cord.MethodsSpinal cords of rats after laminectomy at T8/9 were divided in 3 groups randomly: 10 rats were transplanted with SCNC(Group A);10 rats only made models(Group B);10 rats were as normal control group(Group C).All rats received behaviour survey and inclined plane test every week 8 weeks after operation.6 weeks after operation,they were measured with cortical somatosensory evoked potential(CSEP).And 8 weeks after operation,all specimens were studied histologically.ResultsCSEP could be recorded in group A and C,but not in group B.There were marked differenties between group A or C and group B in behavior survey,inclined plane test.Amount of nerve fibers regenerated through the injury gap in group A was significantly larger than that in group B.ConclusionSCNC could facilitate axonal regeneration and promote the repairing of injuried spinal cord.

Objective To explore the effect of different processing and drying methods of corn and hot pepper on fluorine content in coal-burning type of the endemic fluorosis areas, and to screen food processing and drying methods which meet the quality requirements of grain drying and able to effectively reduce the total fluoride intake of local population. Methods Farmers of endemic fluorosis area in Bijie, Guizhou province were divided into 3 groups: sun-baked drying group, stove drying group with air-tight cover and stove drying group with no cover, 10 households in each group. Corn and fresh hot pepper and samples dried for 2 weeks, or 1, 3, 6-month were collected, and water and fluoride content were detected, and the total daily fluoride intake were calculated in accordance with the "Determination of Water in Food" (GB/T 5009.3-2003) and "Determination of Fluorine in Foods"(GB/T 5009.18-2003). Results Fluoride content in fresh corn and dried for 2 weeks, or 1, 3, 6-month [of sunbaked drying group: (1.40 ± 0.16), (1.56 ± 0.14), (2.15 ± 0.47), (2.70 ± 0.64), (4.06 ± 1.75)mg/kg, stove drying group with air-tight cover: (1.41 ± 0.16), (2.39 ± 0.56), (4.60 ± 0.97), (8.46 ± 5.55), (11.36 ± 3.60)mg/kg,stove drying group with no cover: (1.40 ± 0.13), (4.69 ± 3.97), (4.47 ± 2.77), (9.65 ± 6.47), (26.12 ± 14.52)mg/kg] and pepper[sun-baked drying group: (5.41 ± 1.61), (16.60 ± 7.62), (32.60 ± 7.88), (50.26 ± 17.60),(240.20 ± 272.49)mg/kg, stove drying group with air-tight cover: (754 ± 2.95), (3238 ± 11.50), (119.18 ± 156.45),(224.00 ± 196.58), (495.70 ± 417.29)mg/kg, stove drying group with no cover: (4.82 ± 1.25), (44.30 ± 13.48),(122.89 ± 66.43), (334.23 ± 166.05), (531.01 ± 397.40)mg/kg] increased with elongation of drying time, and the group difference was significant(F = 44.77, 128.71, 126.87, 41.61, 53.63, 170.63, all P ＜ 0.05), with the largest rate of increase in stove drying group with no cover, and the lowest in sun-baked drying group;fluoride was significantly lower (t = 7.93,63.07,5.36,11.98,55.76,7.45, all P ＜ 0.05) after sample washing;total fluoride intake per person per day was 2.57 mg in local adult when ate washed and sun-baked corn, peppers, the total fluoride intake were 5.92, 8.14 mg when ate the food processed by other two drying methods and washed corn, peppers,respectively. Conclusions In the coal-burning type of fluorosis endemic area, should take appropriate health education measures, and instruct local residents to use sun bake their edible corn and pepper for human consumption, and cultivate a habit of washing corn and pepper before cooking, which can reduce the population total fluoride intake, and control endemic fluorosis.

Objective To master the condition of iodine deficiency disorders (IDD),residents iodine nutritional status and implementation of prevention measures in Gansu Province and to provide a basis for developing control strategies.Methods Thirty primary schools were selected in Gansu Province utilizing cluster sampling methodology in 2011,In each selected school,40 children aged 8-10 were randomly selected for thyroid examination and urine samples were collected from 12 children,at the same time measuring the average daily salt intake of domestic residents by 3 days weighing method.On the spot random urine samples and salt samples were collected from a subset of children included in the study.Three towns near the selected school were selected randomly and random urine samples were collected from 5 pregnant and 5 lactating women in each selected town.One drinking water sample was collected for analysis from different sources in five directions(east,south,west,north and central)in each administrative village with the sampled schools.In areas with centralized water supply,two tap water samples were collected for analysis.Results The water iodine median of 83 water samples was 2.02 μg/L,the range was 0.20-36.92 μg/L,and iodine median of 1199 salt samples was 27.4 mg/kg,iodine level in 89.2％ (74/83) of the water samples was lower than 10 μg/L.The iodized salt coverage rate was 98.0％ (1175/1199),the qualified rate of iodized salt was 90.3％ (1061/1175) and the consumption rate of qualified iodized salt was 88.5％ (1061/1199).Total of 360 salt samples intake of households were collected,per capita daily salt was 8.5 g,and the range was 2.5-17.8 g.Total of 1200 children thyroid were checked,the thyroid goiter rate(TGR)of children was 2.8％(34/1200) by B-ultrasound and 3.5％ (42/1200) by palpation.Total of 359,450 and 450 urine samples of childen aged 8-10,pregnant and lactating women,the urinary iodine median were 216.0,168.6,161.9 μg/L,respectively.Conclusions Great progress has been made in the prevention and control of IDD in Gansu Province.IDD has been controlled effectively and the urinary iodine medians are at optimal levels.

Objective:To study the mechanism of ginsenoside Rh2 inhibiting HepG2 cells migration.Methods:HepG2 cells in logarithmic growth phase were cultured in 96-well plates,which were induced by different concentration Rh2,respectively for 24,48,72 hours.The cell inhibition was detected by Cell Counting Kit.Transwell chambers was used to checked HepG2 cell migration ability;luciferase was tested by Luciferase Reporter Assay system reagent;The expressions of P-ERK,ERK,P-P38,P-38,P-JUK,JUK,MMP3 proteins were detect by Western blot;the expression of AP1,MMP3 gene were detected by Quantitative PCR;The expression of AP1, MMP3 fluorescence protein were observed by fluorescence microscopy.Results:Administrated with different concentration of Rh2 after 24 ,48 ,72 h,the proliferation of HepG2 cells were inhibited ( P<0.05) ,and in dose-and time-dependent manner.Transwell assay showed Rh2 could significantly inhibited migration of HepG2 cells.The expressions of P-ERK , MMP3 proteins were significantly decreased,the expressions of P-JUK, P-P38 proteins were significantly increased, expression levels of ERK, P-38, JUK were no significant difference.Expression of AP1,MMP3 gene were significantly decreased,the expressions of AP1,MMP3 fluorescence proteins were significantly decreased.Conclusion:Ginsenoside Rh2 can activate MAPK pathway to inhibit the migration of HepG2 cells.

Objective: To investigate the inhibitory effect of Rh2 on HepG2 cells and explore the underlying mechanism.Methods: We used lentivirus carrying β-catenin to infect HepG2 cell, and detected expression of β-catenin using fluorescence microscopy.The effect of Rh2 on proliferation of HepG2-β-catenin and HepG2 cells was measured by CKK-8 assay,and flow cytometry was used to detect cell cycle and apoptosis.The activity of Gsk-3βwas checked by ELISA kit.The expression of Gsk-3β,β-catenin,Bax,Bcl2,CyclinD1,MMP3 genes were measured by qRT-PCR.In order to checked the relationship between β-catenin and TCF4,CHIP assay kit was used,the expression of Bax,Bcl2,CyclinD1,MMP3 genes were measured by PCR.The expressions of Gsk-3β,β-catenin,Bax,Bcl2,CyclinD1,MMP3 proteins were examined by Western blot.Results:HepG2 cells were successfully infected by pLOV-EF1a-MCS-3FLAG-β-catenin lentivirus,named HepG2-β-catenin.CCK-8 showed that ginsenoside Rh2 could effectively inhibit the proliferation of HepG2 and HepG2-β-catenin cells in vitro,which exhibits a dose-dependent manner at range of 10-160 μmol/L Rh2.The IC50 of Rh2 exposure on HepG2 cell for 48,72 h were 100 μmol/L and 58.12 μmol/L,but the IC50 of Rh2 exposure on HepG2-β-catenin for 48,72 h were 129.2 μmol/L,83.33 μmol/L,respectively.The IC50 of Rh2 exposure on HepG2-β-catenin cell was higher than HepG2 cell, compared with HepG2 group the differences was statistically significant ( P<0.01 ).Flow cytometry indicated that Rh2 could arrest HepG2 and HepG2-β-catenin cells in G0/G1 phase;the cell population in G0/G1 phase of HepG2+Rh2 group was(64.57±0.65)%,HepG2-β-catenin+Rh2 group was(58.61±2.01)%.Flow cytometry indicated that Rh2 could induced early apoptosis in HepG2 and HepG2-β-catenin cells.The apoptosis rate of HepG2 +Rh2 group was (17.27 ±2.77)%,HepG2-β-catenin +Rh2 group(9.02 ±1.76)%.The ELISA results indicated that HepG2 cells was induced by Rh2 for 12,24,48,72 h,the activity of Gsk-3βgradually increased,peak in 48 h,then decreased.Compared with control group,Rh2 induced HepG2 and HepG2-β-catenin cells for 48 hours, Gsk-3βactivity were increased, and their activity reduced after adding Bio, there were no significant differences between HepG2+Rh2 and HepG2-β-catenin+Rh2 groups.The PCR,CHIP and WB results showed that the expression of Gsk-3β,Bax gene and proteins increased,while theβ-catenin,CyclinD1,Bcl2,MMP3 gene and proteins down-regulation in HepG2 and HepG2-β-catenin cell induced by Rh2.Compared with HepG2-β-catenin +Rh2 group, the expression of other gene and proteins changed significantly,however,Gsk-3βwas no significant difference.Conclusion:Over-expression of β-catenin may weaken the phar-macological effects of ginsenoside Rh2 on HepG2 cells.The activity of Gsk-3βwas increased by ginsenoside Rh2 to degradeβ-catenin, affecting the expression of downstream genes,promoting apoptosis of liver cancer cells and inhibiting metastasis.