Objective To investigate the effect of applying scenario simulation practice in the professional ICU nursing students' emergency and intensive care intensified skill training before internship.Methods Totally 120 professional ICU nursing students from grade 2010 and 2011 were enrolled.Control group included 60 students from grade 2010 and experimental group included 60 students from grade 2011.Studnets in control group accepted standardized routine training while those in experimental group accepted scenario simulation practice.All students participated in skill examination after training.Two questionnaires were designed to survey nursing students and clinical teachers respectively at one month after internship.One questionnaire was to evaluate students' attitude to the training and the other one was to survey clinical teachers' evaluation of students' abilities.T test was used to statistically analyze skill examination performance and questionnaire scores between two groups.Results Skill examination performance of experimental group was better than that of control group (P＜0.01).Effects of intensified training were better in experimental group than in control group (P＜0.05,P＜0.01) including the aspects of combining theory and practice,shortening internship role transition time,enhancing selfconfidence of clinical practice and relieving tension anxiety before practice,etc.Clinical teachers thought that the experimental group's analysis ability,strain capacity,operation ability,communication ability,team cooperation ability and actively acquiring knowledge ability were stronger than those of control group (P ＜ 0.01).Conclusions Scenario simulation practice is helpful to improve operation ability and comprehensive ability.Nursing students can adapt to the clinical work better.Findings may be beneficial to the improve-ment of teachers' professional level.

Abstract: Objective To analyze the genetic evolution and molecular characteristics of the H5N6 avian influenza virus A/Changsha/1/2022(H5N6) in a human infection case in Changsha City, providing insights for the prevention and control of H5N6 avian influenza transmission to humans. Methods The sample underwent whole-genome sequencing on a third-generation sequencing platform. Reference sequences were downloaded from the NCBI and GISAID databases for comparison. The phylogenetic tree was constructed and key amino acid mutation sites were analyzed using MEGA7 software. Results The sequence analysis results revealed that the strain studied belongs to the H5 subtype Clade 2.3.4.4b branch. The PB1 gene shared 99.69% homology with the H5N6 virus A/duck/Hunan/S40199/2021(H5N6) from Hunan Province; PB2 had 98.58% homology with A/Whooper swan/Sanmenxia/Y36/2020(H5N8); and other sequences showed high homology with the strain A/Guangdong/1/2021(H5N6) discovered in Guangzhou in 2021. These results indicate that an infection with a recombinant H5N6 subtype avian influenza virus caused the case. The amino acid composition of the cleavage site was RERRRKR↓GLF, consistent with high-pathogenicity avian influenza characteristics. The HA sequence showed no mutations at the Q226L and G228S sites, indicating the preservation of characteristics binding to avian receptors. However, mutations at the S127P, S137A, T160A, T192R, and A267T sites increased the virus's affinity for humans. A deletion found in positions 59-70 of the NA protein stalk implies enhanced viral virulence in mammals. Mutations were noted in the internal genes; S622G in PB1, K389R and V598T in PB2, N409S in PA, N30D and T215A in M1, and P42S in NS1, which may enhance the virulence of avian influenza virus in infecting mice. Conclusions The H5N6 virus strain detected in a human case in Changsha City in this study is highly pathogenic, prone to bind to avian receptors, but numerous mutations at key amino acid sites facilitate infection in humans. Continuous monitoring and research on H5N6 avian influenza virus should be strengthened

Objective:To evaluate the application of analyzing neutrophil cell surface marker CD64 in diagnosis of premature infants infection.Methods:109 infants inpatient in neonatal department(including NICU)were enrolled in the study.CD64 was measured by FCM,which was compared with C-reactive protein(CRP)and IL-6.Results:There was a statistically significant difference in quantitation of CD64 on neutrophil cells(P

BACKGROUND:Osteoporosis caused by diabetes melitus as common secondary osteoporosis has been paid more and more attention recently. Zoledronic acid serves as a novel drug for osteoporosis, and its effect on osteoblasts in vivo remains unclear. OBJECTIVE:To investigate the changes of the expression of bone morphogenetic protein 2 andNoggin in the femur of type 1 diabetes melitus rats and the effect of zoledronic acid on them. METHODS:Models of type 1 diabetes melitus were established by intraperitoneal injection of streptozotocin in 130 Wistar rats. 3 days later, rats with blood sugar > 16.7 mmol/L for three consecutive times were considered as successful models, 120 in total. These models were randomly divided into model, prevention and treatment groups. Rats in the prevention and treatment groups were intravenously administered zoledronic acid (0.1 mg/kg) on the day of modeling and 2 weeks after model establishment. An additional 40 rats were injected with citrate buffer solution as control group. RESULTS AND CONCLUSION: Compared with the control group, femur bone mineral density, serum alkaline phosphatase levels, and femur bone morphogenetic protein 2 mRNA expression levels were significantly lower in the model group (P < 0.05), butNoggin mRNA expression significantly increased (P < 0.05). Compared with the model group, bone mineral density and bone morphogenetic protein 2 mRNA expression levels were significantly higher in the prevention and treatment groups (P < 0.05), butNoggin mRNA expression significantly lower (P < 0.05), and serum alkaline phosphatase levels gradualy restored. These results indicated that the bone metabolic disturbance occurs in early stage in rats with type 1 diabetes melitus. Zoledronic acid can promote bone formation, increase bone density, and improve bone metabolism.

Objective To study the temporo-spatial changing laws of calcitonin gene-related peptide(CGRP)in the rat model of sciatic nerve ligation.Methods Totally 48 Sprague-Dawley rats were randomly divided into sham-operation group(n=6)and experimental groups which survived for 1,3,5,7,14,21,28 d respectively after sciatic nerve ligation(n=6 at each time point).The distribution and quantity of CGRP in sciatic nerve,dorsal root ganglion(DRG)and spinal cord were detected by immunohistochemistry and image analysis methods.Results A large amount of CGRP piled up in the sciatic nerve at 1 d after the ligation,significantly higher in proximal segment than that in distal segment,gradually dropping till disappear basically at 14 d in proximal segment and at 7 d in distal segment.The expression of CGRP in DRG at 7 d began to drop after the ligation,at 21 d to the minimal value and at 28 d lower than normal.The CGRP-immunoreactivety in ipsilateral spinal dorsal horn at 14 d decreased after operation,but the immuno-positive areas of CGRP were of no changes.There are no changes in the CGRP-positive spinal ventral motoneurons of all groups.Conclusion The changes of CGRP presented a temporo-spatial pattern following peripheral nerve legation,maybe resulting from the deficiency of neurothrophins from target organs.

Objective To verify the targeting regulatory relationship between microRNA -939 (miR -939) and CD2 -associated protein (CD2AP).Methods The online RegRNA software was used to predict the human CD2AP promoter for potential binding sites complementary to miR -939.HEK -293T cells were cotransfected with hu-man CD2AP promoter plasmid pGL3 -2K and microRNA negative control (miR -NC)or miR -939 mimics,and the relative luciferase activity(RLA)was detected at 24 h post -transfection.HEK -293T cells were transfected with miR -NC or miR -939 mimics for 48 h,and the CD2AP mRNA expression level was detected by adopting reverse tran-script and real -time fluorescence quantification -PCR,while the CD2AP protein expression level was detected by using Western blot.Results (1 )There were 2 miR -939 binding sites at CD2AP promoter region,located at -468 to -491 and -654 to -677 upstream of initiation codon ATG (marked as +1 )relatively.(2)At 30 nmol/L,50 nmol/L,the RLA in miR -NC group and miR -939 group were 6.81 ±0.88 vs 6.07 ±2.24,5.88 ±1 .44 vs 3.94 ± 0.79 relatively,and there were no significant differences between the 2 groups (t =3.04,2.06,all P >0.05),while the RLA between the 2 groups were 5.58 ±0.58 vs 3.29 ±0.64 at 1 00 nmol/L,and the difference was significant between the 2 groups(t =4.07,P <0.05).(3)At 30 nmol/L,50 nmol/L and 100 nmol/L,the relative CD2AP mRNA expression in miR -NC group and miR -939 group were 1 .00 ±0.01 vs 0.80 ±0.08,1 .00 ±0.00 vs 0.80 ±0.1 3 and 1 .00 ± 0.00 vs 0.72 ±0.07 relatively,while the CD2AP mRNA expression was decreased by 20% -30% at each concentration level,and there were significant differences between the 2 groups (t =4.44,2.93,6.84,all P <0.05).(4)At 50 nmol/L, the relative CD2AP protein expression in miR -NC group and miR -939 group were 0.48 ±0.09 vs 0.19 ±0.12,and the CD2AP protein expression was decreased,and the difference was significant (t =3.36,P <0.05).Conclusions CD2AP is the target gene of miR -939,and miR -939 can down -regulate the expression of CD2AP both in mRNA and protein levels by targeting its promoter region,which indicates that miR -939 may mediate the podocyte injury.

Objective To discuss the treatment effect and application value of salmon calcitonin nasal spray in the patients with postmenopausal osteoporosis.Methods 130 patients with postmenopausal osteoporosis were divided into observation group and control group by random number method,65 cases in each group.The control group was given calcium and alfacalcidol treatment,the observation group received in combination with salmon calcitonin nasal spray treatment.The clinical effect of the two groups was recorded.Results The total effective rate of the treat-ment group was 95.38%,which of the control group was 84.62%,the difference between the two groups was statisti-cally significant (χ2 =4.188,P<0.05 ).After treatment,the tuberosity bone mineral density,femoral neck BMD, L1 -4 lumbar spine bone mineral density of the observation group were (0.69 ±0.15)g/cm2,(0.77 ±0.21)g/cm2, (0.98 ±0.26)g/cm2,which of the control group were (0.52 ±0.08)g/cm2,(0.64 ±0.13)g/cm2,(0.81 ± 0.15)g/cm2,the differences between the two groups were statistically significant (t=8.062,4.243,4.566,all P<0.05).After treatment,the calcitonin,beta collagen fragment,osteocalcin of the observation group were (2.97 ± 0.86)ng/L,(0.11 ±0.01)ng/mL,(7.18 ±1.14)ng/mL,which of the control group were (2.41 ±0.43)ng/L, (0.35 ±0.08)ng/mL,(15.89 ±3.53)ng/mL,the differences between the two groups were statistically significant (t=4.695,24.000,18.930,all P<0.05).Conclusion Salmon calcitonin nasal spray in the treatment of patients with postmenopausal osteoporosis can reduce pain,inhibit osteoclast activity,promote bone formation,prevent bone loss and increase bone mass,it is safe and worthy of popularizing in clinical application.

Objective To analyze the differential gene expression profiling of liver in rats subjected to hemorrhagic shock(HS) and sham hemorrhage shock(SHAM) by gene chip technology, thus to evaluate the possible molecular pathogenesis of HS. Method 20 male Wistar rats were randomly divided into a SHAM group and a HS group, with 10 rats in each group. Hepatic gene expression profiles were detected by oligonucleotide microarrays of 5705 mouse genes in two groups for three times. Genes with ratio(R) > 2 were identified as up-regulated and R < 0.5 were identified as down-regulated. Biological function of differentially expressed genes was analyzed and 9 genes were selected to undergo semi-quantitative RT-PCR. Results Among the total 5705 probes detected,86 genes showed differential expression in HS group comparison with SHAM group. The expression levels of 72 genes were up-regulated while those of 14 genes were down-regulated significantly. Differentially expressed genes were classified according to their biological function: transport genes, transcription regulator genes, signaling genes, response to stress genes, metabolic genes, development genes and cell adhesion genes. Conclusions cDNA microarray is an efficient and high-throughout method to survey gene expression profiles in HS.The variation of those gene expressions might be a potential pathogenic mechanism for HS that may offer a novel target for further study of therapeutic strategies of HS.

Objective To investigate the correlation of plasma interleukin ( IL)-17 level and IL-17 receptor (IL-17R) expression in the thymus of patients with myasthenia gravis (MG).Methods The blood samples of 63 patients (38 with glucocorticoid treatment, 25 with thymus removal) who admitted to Henan Provincial People′s Hospital between 2010 and 2014 were collected at three different stages: pre-treatment, 1 week post-treatment and 1 month post-treatment.The blood samples of 42 healthy controls were also collected.Enzyme linked immunosorbent assay was used to evaluate the levels of IL-17 in plasma.Twenty-five thymus tissues from MG patients and another 12 thymus tissues from patients with congenital heart disease who had surgery therapy were also collected.Reverse transcription polymerase chain reaction was used to evaluate the mRNA levels of IL-17R.The possible correlation between the expression of IL-17 and IL-17R with MG was analyzed.Results Before treatment, the levels of IL-17 in the plasma were much higher in all the MG patients ( both ocular and generalized) when compared to the healthy controls ( controls (3.2 ±0.7) pg/ml, MG patients (8.5 ±1.7) pg/ml, t =2.450, P <0.01; generalized type patients (9.7 ±1.4) pg/ml, t =2.532, P <0.01).In the patients with glucocorticoid treatment, IL-17 levels began to reduce after 1 week treatment and a statistically significant difference was found when compared to the pre-treatment samples (pre-treatment (8.3 ±1.2) pg/ml, 1 week after treatment (6.3 ±0.7) pg/ml, t=2.052, P<0.05) and healthy controls (t =1.933, P<0.05).One month after the glucocorticoid treatment, the levels of IL-17 decreased to the normal level (1 month after treatment (3.9 ±0.6) pg/ml, t=2.630, P <0.01, compared to the pre-treatment; t =1.395, P >0.05, compared to the healthy controls).In the surgery therapy cases, the IL-17 levels were also reduced after the thymus removal ( pre-surgery (8.8 ±1.4) pg/ml, 1 week after surgery (5.3 ±0.7) pg/ml, t=1.950, P<0.05;1 month after surgery (3.0 ±0.4) pg/ml, t=2.683, P<0.01).In the thymus tissues of the MG patients, the mRNA levels of IL-17R were much higher than that of the controls ( relative level 2.31 folds, t =2.682, P <0.01).Meanwhile, a positive correlation was found between the plasma IL-17 levels and the relative IL-17R levels in thymus tissues ( r =0.945 4, P <0.01 ).Furthermore, IL-17 was positively correlated with quantitative myasthenia gravis scores (QMGS) either pre-treatment (r =0.798 1, P <0.01) or post-treatment (r=0.906 5, P<0.01).And IL-17R was positively correlated with QMGS pre-treatment (r=0.775 5, P<0.01).Conclusions IL-17 is increased in the plasma of MG patients (both ocular and generalized) , and is decreased upon the glucocorticoid treatment or surgery therapy, suggesting that it can be used as a parameter to determine the therapeutic effects.IL-17R is increased in the thymus tissues of MG patients, suggesting that it can potentially be used as a pathological diagnosis parameter.

In order to enhance gamma-aminobutyric acid production from L-glutamate efficiently, we amplified the key enzyme glutamate decarboxylase (GAD) encoding gene lpgad from the strain Lactobacillus plantarum GB 01-21 which was obtained by way of multi-mutagenesis and overexpressed it in E. coli BL21. Then we purified GAD by Ni-NTA affinity chromatography and characterized the enzyme to optimize the conditions of the whole-cell transformation. The results showed that the recombinant E. coli BL21 (pET-28a-lpgad) produced 8.53 U/mg GAD, which was increased by 3.24 fold compared with the GAD activity in L. plantarum. The optimum pH and temperature of the enzyme were pH 4.8 and 37 degrees C, respectively. At the same time, we found that Ca2+ and Mg2+ could increase the activity significantly. Based on this, we investigated gamma-aminobutyric acid transformation in 5 L fermentor under the optimum transformation conditions. Accordingly, the yield of gamma-aminobutyric acid was 204.5 g/L at 24 h when the 600 g L-glutamate was added and the mole conversion rate had reached 97.92%. The production of gamma-aminobutyric acid was improved by 42.5% compared with that under the unoptimized transformation conditions. This paved a way for the gamma-aminobutyric acid construction of the industrial applications.
Cloning, Molecular ; Escherichia coli ; enzymology ; genetics ; metabolism ; Glutamate Decarboxylase ; biosynthesis ; genetics ; Glutamic Acid ; metabolism ; Lactobacillus plantarum ; enzymology ; genetics ; Recombination, Genetic ; gamma-Aminobutyric Acid ; biosynthesis