Objective:To investigate the expression and phosphorylation level change of adenosine monophosphate activated protein kinase (AMPK) in skeletal muscle of severely scald rats and its roles in skeletal muscle atrophy in severely scalded rats.Methods:The experimental research method was applied. Totally 100 6-week-old male Wistar rats were divided into sham injury group and scald group according to the random number table, with 50 rats in each group. After weighing the body weight, rats in scald group were inflicted with full-thickness scald of 30% total body surface area on the back, and rats in sham injury group were simulated with scald. At 6 h and on 1, 3, 5, and 7 d post injury, 10 rats in each group were taken to measure their body weights and weights of extensor digitorum longus and soleus muscle. At 6 h and on 1, 3, 5, and 7 d post injury, the tibialis anterior muscles were collected, the mRNA expressions of muscle atrophy F-box protein (MAFbx) and muscle-specific RING finger protein 1 (MuRF1) were detected by real-time fluorescent quantitative reverse transcription polymerase chain reaction; the content of adenosine monophosphate (AMP), adenosine diphosphate, and adenosine triphosphate (ATP) were detected by high performance liquid chromatography, and AMP/ATP ratio and energy charge were calculated; the protein expressions of AMPK-α and phosphorylated AMPK-α (p-AMPK-α) were detected by Western blotting, and the p-AMPK-α/AMPK-α ratio was calculated, with sample number of 4 in each time point of each group. Data were statistically analyzed with analysis of variance for factorial design and least significant difference test.Results:The body weights of rats in 2 groups before injury and at each time point post injury were close ( P>0.05). At 6 h post injury, the weight of extensor digitorum longus of rats in scald group was (0.107±0.007) g, which was significantly heavier than (0.086±0.0607) g of sham injury group ( P<0.01). On 3 d post injury, the weight of extensor digitorum longus of rats in scald group was (0.083±0.016) g, which was significantly lighter than (0.102±0.005) g of sham injury group ( P<0.01). The weight of soleus of rats in 2 groups were close at each time point post injury ( P>0.05). Compared with those of sham injury group, the mRNA expression of MAFbx in tibialis anterior muscle of rats in scald group was significantly up-regulated at 6 h post injury ( P<0.01), and the mRNA expressions of MuRF1 in tibial anterior muscle of rats in scald group were significantly up-regulated at 6 h and on 1 d post injury ( P<0.01). At 6 h and on 7 d post injury, compared with those of false injury group, the AMP/ATP ratios of the tibial anterior muscle of rats in scald group were significantly increased ( P<0.05 or P<0.01), and energy charges of the tibial anterior muscle of rats in scald group were significantly decreased ( P<0.01). At each time point post injury, the protein expressions of AMPK-α of the tibial anterior muscle of rats in 2 groups were close ( P>0.05). The p-AMPK-α/AMPK-α ratios of the tibial anterior muscle of rats in scald group at 6 h and on 7 d post injury were significantly higher than those in sham injury group ( P<0.05 or P<0.01). Conclusions:The decrease in energy charge and increase in AMP/ATP ratio of skeletal muscle of rats after severe scald activate AMPK. The activation of AMPK in the early stage of injury is consistent with the up-regulation of MAFbx and MuRF1 expressions and down-regulation of skeletal muscle weight. The above-mentioned changes may be one of the molecular mechanisms of skeletal muscle atrophy in rats with severe scald

Objective To investigate the application and effect evaluation of combined warming and humidification device in patients with artificial airway in high altitude area. Methods The convenient sampling method was used to select 225 hospitalized patients in the Department of Critical Care Medicine of the People′s Hospital of Tibet Autonomous Region from January to June 2018. According to the order of admission time, they were divided into two groups: observation group (125 cases) and control group (100 cases). The observation group was humidified by artificial nose, while the control group was humidified by heating and humidifying device. The phlegm properties and scab formation were observed in the two groups. The ambient temperature, humidity and airway temperature were measured at 3:00, 9:00, 15:00 and 21:00 every day. The average values of four measurements were taken. The levels of CO2 partial pressure and oxygen partial pressure were recorded by blood gas analysis. χ2was used to compare the differences between the 2 groups. Results There was no difference in sputum properties between the two groups before humidification. The incidence of grade II and grade III mucous phlegm in the observation group was 46, 48 and 65 on the first, second and third day after humidification, while that in the control group was 13, 9 and 10 respectively. The difference between the two groups was statistically significant (χ2=16.266, 25.387, 44.100, P<0.01). On the 2nd and 3rd day, the oxygen partial pressure levels of the observation group were (92.62 ± 5.73), (91.34 ± 4.82) mmHg, and those of the control group were (96.17 ± 3.60), (95.53 ± 2.96) mmHg, respectively. The difference between the two groups was statistically significant (t=5.697, 8.045, P<0.01). The sputum scab formation and airway temperature in the observation group were 41 cases and(29.89±1.95)℃ respectively after 3 days of humidification, 7 cases and(34.79±1.82)℃respectively in the control group. The difference between the two groups was statistically significant (χ2= 22.035, t =- 26.031, P < 0.01). Conclusion The combination of heating and humidifying device can effectively improve the sputum properties, reduce the formation of sputum scab, effectively improve the level of carbon dioxide, without increasing the workload.

Objective:To study the feasibility of 3-D reconstruction model in the observation of tetrachlorodibenzo-p-dioxin(TCDD) effected palatal organ development of fetal mouse.Methods:Kunming mice treated 40 ug/kg TCDD by lavage on day 12.5 of pregnancy were used as in the experimental group,isodose corn oil treated in the control group.On day 13.5,14.5 and 15.5 of pregnancy heads of the fetal mice were taken out and fixed.Conventional paraffin serial sections of palatal organ were preparated and dyed by hematoxylin-eosin,images of the palatal organs were collected and photoshop treated,3-D reconstruction of the palatal organ was performed by 3D-DOCTOR software.Results:3-D reconstruction images showed that palatal organs moved from on both sides above the tongue and gradually closed and merged in the control group.In the experimental group,the palatal organs moved from on both sides above the tongue was later than control group,gradually closed,but not merged,formed cleft palate.Conclusion:3D-DOCTOR software reconstruction can be used for the study of the development process effected by TCDD in the pregnant mouse.

OBJECTIVE: To determine the mass scores of naringin and neohesperidin in Fructus aurantii and Citrus chang-shan-huyou with their processed products and evaluate the quality of Fructus aurantii and Citrus changshan-huyou with their pro-cessed products. METHODS:According to the requirements of Chinese Pharmacopoeia(2010 edition)and Zhejiang Province Tradi-tional Chinese Medicine Preparation Standards (2005 edition),the moisture and ash of F. aurantii and C. changshan-huyou with their processed products were detected. And the contents of naringin and neohesperidin were determined. The ZORBAX SB-C18 column was used with the mobile phase of acetonitrile-water(20∶80,V/V)at the flow rate of 1.0 ml/min. The detection wave-length was set at 283 nm,and the column temperature was 40℃.The samples size was 10μl. RESULTS:The moisture of F. au-rantii and C. changshan-huyou was decreased after processing with no obvious change for ash. The contents of naringin and neohes-peridin were decreased,significantly for F. aurantii,and all consistent with the requirements of Chinese Pharmacopoeia(2010 edi-tion)except F. aurantii. The linear range was 0.028 45-0.284 5μg(r=0.999 7)for naringin and 0.085 9-0.858 6μg(r=0.999 6)for neohesperidin;the RSDs of precision,stability and reproducibility tests were no more than 1.36% and the average recovery was re-spectively 96.45%-100.43%(RSD=1.45%,n=6) and 98.36%-102.00%(RSD=1.26%,n=6). CONCLUSIONS: There was no significant difference in the inspection and determination re-sults in F. aurantii and C. changshan-huyou. It is suggested to adjust the limitation of content determination in the Chinese Pharmacopoeia(2010 edition)and processed standards.

Objective To investigate the effect of kangaroo mother care on children′s comfort with intramuscular injection. Methods One hundred children with an intramuscular injection were grouped according to the injection date. Those treated with injection on odd-numbers days were assigned as the intervention group , and those with injection on even-numbered days as the control group, with 50 cases in each group. The control group received routine care and the intervention group with kangaroo mother care. The two groups were compared in terms of comfort level. Result The comfort level of the observation group was higher than that of the control group with statistical significance (P<0.01). Conclusion Kangaroo mother care can improve the comfort level of children patients with intramuscular injection. It is a simple, safe and effective in non-pharmaceutical pain therapy.

Objective To study the gene homology of intestinal colonized and infectious bacteria in ICU patients to provide the epidemiological and molecular biological basis for formulating the control measures of multi resistant bacterial hospital infection. Methods The multi-drug resistant colonized bacteria isolated from the anal swabs and the same multi-drug resistant bacteria isola-ted from the clinical samples in the same patients were matched.The Diversilab automatic repetitive extragenic palindromic(REP)-PCR typing system was adopted to analyze the gene homology of multi-drug resistant colonized bacteria and infectious bacteria in the intestinal tract.Results 4 pairs of multi-drug resistant colonized bacteria and the same multi-drug resistant bacteria isolated from the clinical samples on admission in the same patients were selected and performed the homology detection,2 pairs had the ho-mology and 2 pairs had no homology;4 pairs of multi-drug resistant colonized bacteria and the same multi-drug resistant bacteria isolated from the clinical samples in the patients with hospital infection were performed the homology detection,4 pairs all showed the homology.Conclusion The multi-drug resistant colonized bacteria and the infectious bacteria in ICU patients have the homolo-gy.The multi-drug resistant colonized bacteria can cause the occurrence of hospital infection,so their management should be strengthened in clinic.

OBJECTIVETo detect mutations of the retinitis pigmentosa GTPase regulator (RPGR) gene in two Chinese X-linked retinitis pigmentosa families.

METHODSFragments of exons 1-19 of the RPGR gene were amplified with intronic primers, using genomic DNA as template. The polymerase chain reaction (PCR) products were analysed by single-strand conformation polymorphism (SSCP) and direct sequencing. Mutations were identified by comparing DNA sequences of the patients with those of the normal controls.

RESULTSTwo novel mutations, c1536delC and E332X, were identified in exons 12 and 9 of the RPGR gene in both families. Each mutation was the first mutation found in their respective exons. Both mutations were predicted to cause premature termination, which resulted in truncated proteins without normal functions of the RPGR products.

CONCLUSIONSBoth mutations are the genetic basis of the pathogenesis in the respective families. Our data might be helpful in analysing the function of the RPGR protein.

Carrier Proteins ; genetics ; Eye Proteins ; Female ; Genetic Linkage ; Humans ; Male ; Mutation ; Polymerase Chain Reaction ; Polymorphism, Single-Stranded Conformational ; Retinitis Pigmentosa ; genetics ; Sequence Analysis, DNA ; X Chromosome

AIM: To investigate the protective effects and mechanisms of iptakalim hydrochloride(Ipt)on H_2O_2 induced neurotoxity. METHODS: Neurotoxity injury was induced by H_2O_2 in PC12 cells. The cell viability was tested by MTT assay. The glutamate released from PC12 cells was measured by HPLC combined with fluorescent detector analysis. Changes in the intracellular free Ca 2+ concentration ([Ca 2+ ]_i) were determined in fluo-3 AM loaded PC12 cells. RESULTS: Ipt (1, 10 and 100 ?mol?L -1 ) markedly mitigated H_2O_2-induced neurotoxity, 10 ?mol?L -1 Ipt inhibited the release of glutamate and the increase of [Ca 2+ ]_i induced by H_2O_2 .The protective effects was incompletely blocked by 5-HD which is a mitochondrial K_ ATP channels antagnist. CONCLUSION: Ipt provides neuroprotective effects on H_2O_2 induced cytoxixity in cultured PC12 cells and the protective effects may be partially related with mitochondrial KATP channels.

Objective To explore the relationship between the genetic markers of the candidate genes (insulin receptor, glucose transporter 2 and glucokinase) and non-insulin dependent diabetes mellitus (NIDDM) in Chinese pedigrees.Methods Genomic DNA of 87 members in 12 Chinese NIDDM pedigrees was amplified using PCR technique. Denaturing polyacrylamide gel electrophoresis was applied to detect short tandem repeat polymorphisms. The linkage program was used under the models of autosomal dominant and recessive. The individuals with impaired glucose tolerance (IGT) of age ≤45 and age >45 were counted as affected and unknown status respectively.Results Linkage of insulin receptor gene and NIDDM was strongly and significantly rejected with total LOD score of -∞ (θ=0.000). Linkage analysis of glucose transporter 2 gave the result with total LOD score of 2.646 (θ=0.000) in 4 pedigrees. Linkage of glucokinase and NIDDM was of significance with total LOD score reaching 3.150 in the integration of 6 pedigrees.Conclusions Mutations in or near the insulin receptor gene are unlikely to be the major cause of inherited predisposition to NIDDM in Chinese pedigrees that are related in this study, not excluding the principal role in the other Chinese or populations, while the mutations/variations near or at the glucose transporter 2 gene locus might be something of the cause, which needs more data to be ascertained, of familial NIDDM in Chinese pedigrees. However, the variations in or near the glucokinase gene might play a dominant role in the development of NIDDM in some Chinese and represent a subgroup of common NIDDM.

Objective: To study the meal qanity of the patient with chronic renal failure(CRF). Methods: The nutritional state of 140 CRF patients was researched,basing on the diet history. Results: With the progression of course of CRF ,the nutritional states of patients with CRF were getting worse . Conclusions: The nutritional state of CRF patients should be paid attention to.