Objective To observe the changes to,and possible mechanism of,intestinal permeability in pigs without direct injury after an abdominal-and intestinal-penetrating injury from firearms in cold environment at high altitudes.Methods Fifty-five experimental pigs were divided into two groups:high-altitude cold group(HC)and low-altitude normal temperature group(LN).According to the observation time,each group was divided into five experimental subgroups:0h,2h,4 h,8h,and 24 h.There were six pigs in each HC subgroup and five pigs in each LN subgroup.After euthanasia,intestinal tissues were taken,and the levels of the inflammatory factors TNF-α,and IL-6 in intestinal homogenate and the concentrations of intestinal permeability-related proteins DAO and D-lactate acid in blood were detected by ELISA method.The intestinal tissues of experimental pigs were taken at 0 h and 8 h for LN and 8 h for HC,and intestinal pathological changes were observed and scored after HE staining.The concentrations of Occludin,ZO-1,Claudin-3,TLR4,NF-κB,and MLCK(proteins related to intestinal permeability)were detected by Western blot to explore the effect of a cold environment at high altitude on secondary intestinal permeability changes after injury and the possible mechanisms.Results Both the HC group and LN group experienced typical abdominal intestinal penetrating injuries,and there were no significant differences in their abdominal infection scores or intestinal adhesion(P＞0.05).The levels of DAO and D-LA in the serum of experimental pigs in the HC and LN groups gradually increased over time.The levels of DAO and D-LA in the HC group were significantly higher than those in the LN group at all time points(P＜0.01 or P＜0.001).The fastest increase in DAO and D-LA in the HC group was 4 h to 8 h,while in the LN group,it was 8 h to 24 h.The pathological score of intestinal tissue in the HC group was significantly higher than that in the LN group of experimental pigs(P＜0.01).The inflammatory factors TNF-α and IL-6 both increased over time in the intestinal tissue of LN and HC groups.The most significant time point for a increase of inflammatory factors in the HC group was 4 h to 8 h,while in the LN group,it was 8 h to 24 h.The intestinal tissue IL-6 and TNF-α levels of experimental pigs in the HC group were higher than those in the LN group the entire time(P＜0.05,P＜0.01,or P＜0.001).The levels of occludin and ZO-1 in the HC group at 8 h decreased significantly compared to those of the LN group at the 8 h time point(P＜0.05),while claudin-3 showed a significant decrease in LN(P＜0.001).In the HC group,TLR4,NF-κB,and MLCK were both higher than those in the LN group at 8 h,and the difference was statistically significant(P＜0.05).Conclusions A high-altitude cold environment can lead to a secondary increase in intestinal permeability after abdominal-penetrating firearm injury,and its mechanism may be related to the TLR4/NF-κB/MLCK pathway.

Abstract According to the Healthy China Action Plan, Wuhan gives full play to the role of preventing and controlling student myopia by promoting student health. The primary focus is placed on education in schools, and Wuhan has integrated educational resources to develop a multi-level myopia prevention and control system and service network for school students. The network contains educational adminstrative, schools, families, and professional technical service organizations. By integrating multiple disciplines, Wuhan has built a comprehensive vision health management service system for all students. The Internet and cloud intelligent monitoring facilitated the establishment of a smart vision health management platform for students, which thoroughly and efficiently implemented myopia prevention and control to safeguard students visual health by engaging in education, monitoring, and supervision. The prevention and control of student myopia is a breakthrough for comprehensive healthy development of students. A comparison of the standard myopia rate in Wuhan in 2019 and 2018 revealed that the standard myopia rate at different learning stages of primary school, junior high school, and high school dropped by 3.31, 2.50, and 2.26 percentage points, respectively, and the rate of myopia in primary school was significantly lower than the national level. Post-epidemic surveys showed that the compliance rate and the awareness rate of the visual environment and visual behaviors of primary and secondary school students in Wuhan reached more than 80%, and prevalence of newly onset myopia or decreased vision was 30%, which was lower than the national average. The "Wuhan Model" provides an important referential framework for public health services for school students.

Objective:To evaluate the effect of melatonin on prefrontal cortex ischemia-induced cognitive impairment in rats and to investigate the receptor mechanism.Methods:Clean-grade adult male Sprague-Dawley rats, weighing 300 g, were selected, and a catheter was implanted into the prefrontal cortex.The experiment was performed in two parts.Experiment Ⅰ Twenty-four rats, in which catheters were successfully inserted into the prefrontal cortex, were assigned into 3 groups ( n=8 each) using a random number table method: control group (group C), model group (group M) and melatonin group (group ME). Normal saline 0.5 μl was injected into the prefrontal cortex in group C, 1 μmol/L endothelin 0.5 μl was microinjected into the prefrontal cortex in group M, and 1 μmol/L endothelin and 1 μmol/L melatonin 0.5 μl were injected into the prefrontal cortex in group ME.Experiment Ⅱ Forty-four rats, in which catheters were successfully inserted into the prefrontal cortex, were assigned into 4 groups ( n=11 each) using a random number table method: model group (group M), melatonin group (group ME), MT 1/2R antagonist luzindole + melatonin group (group L + ME) and MT 2R antagonist 4p-pdot + melatonin group (group P + ME). In group M, 1 μmol/l endothelin 0.5 μl was microinjected into the prefrontal cortex.In group ME, 1 μmol/L endothelin + 1 μmol/L melatonin 0.5 μl was injected into the prefrontal cortex.In group L + ME, 1 μmol/L endothelin + 1 μmol/L MT 1/2R antagonist + 1 μmol/L melatonin 0.5 μl was injected into the prefrontal cortex.In group P + ME, 1 μmol/L endothelin + 1 μmol/L MT 2R antagonist + 1 μmol/L melatonin 0.5 μl was injected into the prefrontal cortex.T-maze and the open field tests were performed at 1 week after administration. Results:Experiment Ⅰ There was no significant difference in the locomotor speed in open field test among C, M and ME groups ( P>0.05). The rate of correct selection in T-maze test was significantly lower in M and ME groups than in group C and higher in group ME than in group M( P<0.05). Experiment Ⅱ There was no significant difference in the locomotor speed in open field test among the four groups( P>0.05). Compared with group M, the rate of correct selection in open field test was significantly increased in ME and P+ ME groups ( P<0.05), and no significant change was found in group L+ ME ( P>0.05). Compared with group ME, the rate of correct selection in open field test was significantly decreased in group L+ ME ( P<0.05), and no significant change was found in group P+ ME( P>0.05). Conclusion:Melatonin can attenuate prefrontal cortex ischemia-induced cognitive impairment in the rats, and the mechanism is related to activation of MT 1R.

Objective:To investigate the relationship between lateral hypothalamus and melatonin-induced reduction of wakefulness in rats and the receptor mechanism.Methods:Forty clean-grade adult male Sprague-Dawley rats, weighing 250-300 g, were divided into 4 groups ( n=10 each) using a random number table method: control group (group C), melatonin group (group M), melatonin type-1/2 receptor (MT 1R) antagonist luzindole plus melatonin group (group L+ M), and melatonin type-2 receptor (MT 2R) antagonist 4P-PDOT plus melatonin group (P+ M group). In group C, 0.5 μl of 0.9% NaCl solution was microinjected into the lateral hypothalamus.In group M, 1 μmol/L melatonin 0.5 μl was microinjected into the lateral hypothalamus.In group L+ M, 1 μmol/L MT 1/2R and 1 μmol/L melatonin (0.5 μl in total) was microinjected into the lateral hypothalamus.The microinjection time was from 19: 30 to 20: 00.The changes in sleep-wake duration and the oscillating energy in different frequency bands of electroencephalogram were detected by using electroencephalogram and electromyogram recording technology. Results:Compared with group C, the percentage of wakefulness time was significantly decreased, the percentage of non-rapid eye movement sleep and rapid eye movement sleep time was increased, the energy for delta oscillation was increased, the energy for theta oscillation was decreased, and no significant change was found in the energy for alpha oscillation in M and P+ M groups ( P<0.01), and no significant change was found in the parameters mentioned above in group L+ M ( P>0.05). Compared with group M, the percentage of wakefulness time was significantly increased, the percentage of non-rapid eye movement sleep and rapid eye movement sleep time was decreased, the energy for delta oscillation was decreased, and the energy for theta oscillation was increased in group L+ M ( P<0.01), and no significant change was found in the parameters mentioned above in group P+ M ( P>0.05). Conclusion:The lateral hypothalamus may be involved in melatonin-induced reduction of wakefulness in rats, and the mechanism may be related to activating MT 1R in the lateral hypothalamus.

Objective:To evaluate the effect of propofol on excitability of pyramidal neurons in orbitofrontal cortex of mice and the underlying ion channel mechanism.Methods:Brain slices of 400 μm thickness from healthy male C57 mice (aged 8-12 weeks)were prepared.This experiment was performed in two parts.Part Ⅰ The brain slices were divided into 2 groups ( n=7 each) based on the random number table method: control group (C group) and propofol group (P group). Cells were perfused with vehicle in group C and with 10 μmol/L propofol in group P. Part Ⅱ The brain slices were divided into 5 groups ( n=8 each) using the random number table method: propofol group (P group), hyperpolarization-activated non-selective cation channel antagonist ZD7288 plus propofol group (Z + P group), inward rectifier potassium channel antagonist topiramate plus propofol group(T + P group), transient activation of voltage-gated potassium channel antagonist 4-aminopyridine (4AP) plus propofol group (A + P group), and delayed activation of voltage-gated potassium channel antagonist tetraethylammonium (TEA) plus propofol group (TEA + P group). Cells were perfused with 10 μmol/L propofol for 2 min in P group, with 5 μmol/L ZD7288 and 10 μmol/L melatonin for 2 min in Z+ P group, with 5 μmol/L topiramate and 10 μmol/L propofol for 2 min in T + P group, with 10 μ mol/L 4-aminopyridine and 10 μmol/L propofol for 2 min in A+ P group, and with 10 μmol/L TEA and 10 μmol/L propofol for 2 min in TEA+ P group.The whole-cell currents, membrane potential and discharge frequency of pyramidal neurons in the orbitofrontal cortex were recorded by whole-cell patch-clamp. Results:Part Ⅰ Compared with C group, whole-cell currents were significantly increased, and the membrane potential and discharge frequency were decreased in P group ( P<0.01). Part Ⅱ Compared with P group, no significant change was found in the whole-cell currents, membrane potentials and discharge frequency in Z+ P group, T+ P group and A+ P group ( P>0.05), and the whole-cell currents were significantly decreased, and the membrane potentials and discharge frequency were increased in TEA+ P group ( P<0.05). Conclusion:Propofol can inhibit the excitability of pyramidal neurons in the orbitofrontal cortex, and the mechanism is related to activating delayed activation of voltage-gated potassium channels in mice.

OBJECTIVE: To explore the genetic basis of a fetus with enlargement and enhanced echo of the kidneys. METHODS: The imaging data of the fetus were collected, in addition with 20 mL amniotic fluid sample and 2 mL peripheral blood samples of both parents. Amniotic DNA was extracted for library construction and whole exome sequencing, and Sanger sequencing was carried out to verify candidate variant associated with the fetal phenotype. RESULTS: Prenatal ultrasound showed that the fetus had enlargement and enhanced echo of the kidneys, in addition with many small renal cysts. Whole exome sequencing showed that the fetus carried pathogenic compound heterozygous variants of the ETFDH gene, namely c.3G>C and c.1436dupA. Sanger sequencing of the family suggested that the variants were inherited from its mother and father, respectively. CONCLUSION By combining its clinical manifestations and results of whole exome sequencing, the fetus was diagnosed as glutaric acidemia type ⅡC due to the compound heterozygous variants of the ETFDH gene. Above results have provided a basis for prenatal diagnosis and genetic counseling. Fetal exome sequencing has provided an important tool for prenatal diagnosis.

Objective To evaluate the effects of melatonin on the excitability of pyramidal neurons in the prefrontal cortex and the role of melatonin receptor 1 (MT1 R)-cyclic adenosine monophosphate (cAMP)-protein kinase A (PKA) signaling pathway.Methods Brains were obtained from male SpragueDawley rats between 14 and 21 days after birth.The brain slices of 350-μm thick were prepared and placed in artificial cerebrospinal fluid.The brain slices were divided into 5 groups (n =6 each) using a random number table method:control group (C group),melatonin group (M group),MT1/2R antagonist luzindole plus melatonin group (L+M group),MT2R antagonist 4P-PDOT plus melatonin group (P+M group) and PKA inhibitor Rp-cAMPS plus melatonin group (R+M group).Cells were perfused for 2 min with artificial cerebrospinal fluid in group C.Cells were perfused for 2 min with 1 μmol/L melatonin in group M.Cells were perfused for 2 min with the mixture of 1 μmol/L MT1/2R antagonist luzindole and 1 μmol/L melatonin in group L+M.Cells were perfused for 2 min with the mixture of 1 μmol/L MT2R antagonist 4P-PDOT and 1 μmol/L melatonin in group P+M.In group R+M,1 mmol/L PKA inhibitor Rp-cAMPS was continuously added to the pipette solution,and cells were perfused for 2 min with 1 μmol/L melatonin.The whole-cell patch-clamp technique was used to record the membrane potential and clamp current of pyramidal neurons in the prefrontal cortex.Results Compared with group C,the clamp current was significantly increased,and the membrane potential was decreased in group M (P＜0.05).Compared with group M,the clamp current was significantly decreased,and the membrane potential was increased in L + M and R + M groups (P＜0.05),and no significant change was found in the clamp current or membrane potential in group P+M (P＞0.05).Conclusion Melatonin inhibits the excitability of pyramidal neutrons in the prefrontal cortex,and the mechanism is related to activating MT1 R-cAMP-PKA signaling pathway.

Objective To evaluate the relationship between prefrontal cortex and propofol-induced cognitive dysfunction in rats. Methods SPF healthy male Sprague-Dawley rats, weighing 300-350 g, aged 16 weeks, were used in this study. Thirty rats in which catheters were successfully implanted into the prefrontal lobe were divided into 2 groups ( n=15 each ) using a random number table method: control group (group C) and propofol group (group P). In group P, 50μmol/L propofol 0. 5μl was microinjected into the prefrontal cortex at day 7 after operation, and the equal volume of normal saline was given instead in group C. T-maze test and open field test were performed at 15 min after administration. Results Com-pared with group C, the correct rate of selection in T-maze was significantly decreased ( P＜0. 05) , and no significant change was found in the total locomotor or number of rearing in open field test in group P ( P＞0. 05) . Conclusion Prefrontal cortex may be involved in propofol-induced cognitive dysfunction in rats.

Objective To investigate the long-term effect and radiation side reaction of different boost methods in patients with local remnants of locally advanced nasopharyngeal carcinoma after curative radiotherapy Methods Based on the retrospective analysis , 102 patients with local remnants of locally advanced nasopharyngeal carcinoma, after curative radiotherapy, were enrolled from February 2009 to May 2011. Forty-nine patients received gamma knife boost and 53 patients received 3-dimensional conformal radiotherapy (3D-CRT). The long-term effect and radiation side reaction were compared between the two groups. Results The 1-year, 3-year, and 5-year relapse-free survival rate (RFS) in the gamma knife boost group and the 3D-CRT group was 97.8%, 90.0%, 80.8%, and 94.3%,76.1%,72.9%, respectively (P < 0.05). No significant differences of the overall survival and metastasis-free survival rate wer found in two groups , as well as the long-term radioactive reaction. Conclusion The gamma knife boost is better than 3D-CRT with respect to local control rate , but the long-term radiation side reactions were similar between the methods.

Objective To measure the costs of primary health services in the community for developing government compensation standard and prediction of budget in 201 5.Methods Literature review and panel discussions were used to build the index database.A two-round Delphi expert consultation determined the work to do and steps for community healthcare standards.60 community health centers were sampled by stratified random sampling for survey,and the work hours,workload and service volume of respective services in 2013 were measured according to the service standards of primary care.Results The community primary care consists of the categories of outpatient services,nursing care, laboratory tests and drug management,totaling 20 services and 88 working procedures.The total work hours of primary healthcare service at 60 community health centers were 2 557 187.9 hours,which is adjusted to 2 959 21 5.1 hours based on workload coefficients.Based on the income standards of employees at the community health centers investigated,and the human cost price which was recommended by the experts,the total costs of primary healthcare of the 60 centers were 245 million,248 million and 318 million respectively,and the average cost was 71.0 yuan,71.8 yuan and 92.1 yuan per visit respectively.By such standards,the government should subsidize 38.9 ~ 60.0 yuan per visit to the centers.The total expenditure of primary healthcare of all communities in Shenzhen in 201 5 was predicted to be 314 to 407 million,for which the government is expected to subsidize 1.68~2.58 billion.Conclusion The subsidies for community primary healthcare fall short as the cost per visit runs up in 2013 to 71.0 yuan per visit or more,far above the current subsidy of 32.1 yuan per person.The subsidy per visit should be made 50.0 yuan in 201 5.