Objective To investigate the application of of Gaussian 09/GuassView 5.0 in spectra teaching in Analytical Chemistry.Method Undergraduates of Pharmaceutical Science in 2014 grade of Kunming Medical University were selected to teach with a method with the help of Gaussian 09/GuassView 5.0 soft.Calculations of UV spectra,IR spectra and NMR spectra of compounds were introduced to make better understanding in the learning of relative knowledge points.The teaching effect was evaluated by the comparison of theoretical exam results of 2013 grade which didn't use soft.Result The theoretical test results showed that the scoring averages of the 2014 grade in UV,IR and NMR spectra were significantly higher than that in 2013 grade (P＜0.05).Conclusion Gaussian 09/GuassView 5.0 can visualize the nonobjective knowledge,and imporve the students' interesting,thus improving the efficiency of teaching and learning.

Objective: To evaluate the surgView-RFT electromagnetic navigation system in radiofrequency thermocoagulation(RFT). Methods: 6 foramen ovales of 3 cadavers were punctured by 3 beginners. Every one tried 5 times by free-hand and 5 times by SurgView-RFT electromagnetic navigation-guidance. After each puncture, CT scans were used as golden standard to measure the distance between the tip of needle and the anterior margin of the foramen ovale. T test and variance of SAS 6.12 statistical software was used to do the statistic analysis. Results: The punctures by free-hand failed, and the distances between the tip of needle and the anterior margin of the foramen ovale were 8-10.6 mm. The punctures by navigation guidance were successful, and the distances between the tip of needle and the anterior margin of the foramen ovale were 2.68-3.54 mm. The error of navigation system was 0.59 mm which was verified by CT scan. There was significant difference between free-hand puncture and navigation-guided puncture. Conclusion: The surgView-RFT electromagnetic navigation system is characteristic with high performance, high accuracy and minimal invasion, while it is applied in training or clinical using.

Objective To evaluate the effect of curcumin on lidocaine-induced nerve injury and the role of protein kinase B ( Akt) and extracellular signal-regulated kinase 1∕2 ( ERK1∕2) signaling pathway in rats. Methods Eighty SPF male Sprague-Dawley rats, aged 8-10 weeks, weighing 220-250 g, were di-vided into 8 groups ( n=10 each) using a random number table method: control group ( group C) , sham operation group (group S), lidocaine group ( group L), dimethyl sulfoxide ( DMSO) group, low-dose curcumin group ( group CL ) , high-dose curcumin group ( group CH ) , curcumin plus ERK inhibitor PD98059 group (group P) and curcumin plus Akt inhibitor MK-2206 group (group M). The model of nerve injury was established by injecting 2％ lidocaine 10μl via the catheter in anesthetized rats in the other six groups except for C and S groups. Drugs were injected through a microcatheter for 14 consecutive days starting from 3rd day after IT catheterization as follows: DMSO 10μl, curcumin 100μg∕10μl and curcu-min 500 μg∕10 μl were injected once a day in D, CL and CH groups, respectively; curcumin 500 μg∕10μl was injected once a day, and D9805910μg and MK-220612μg ( in 5μl DMSO) were intraperitoneal-ly injected once a week in P and M groups, respectively. The mechanical paw withdrawal threshold ( MWT) and thermal paw withdrawal threshold ( TWL) on the operated side were measured on 1 day before IT catheterization, before administration on 3rd day after IT catheterization, and on 14th day after adminis-tration. The spinal cord was removed on 1st day after the end of administration for determination of the ex-pression of ERK1∕2, phosphorylated ERK1∕2 (p-ERK1∕2), Akt, phosphorylated Akt (p-Akt), c-fos, Bcl-2 and Bax ( by Western blot) and expression of ERK1∕2 and Akt mRNA ( by real-time polymerase chain reaction) . Results Compared with group C, the MWT was significantly decreased, the TWL was short-ened, the expression of ERK1∕2 protein and mRNA, p-ERK1∕2, c-fos, Akt protein and mRNA, p-Akt and Bcl-2 was down-regulated, and the expression of Bax was up-regulated in group L (P<0. 05). Com-pared with group L, the MWT was significantly increased, and the TWL was prolonged in CL, CH, P and M groups, the expression of p-ERK1∕2, c-fos, p-Akt and Bcl-2 was up-regulated, and the expression of Bax was down-regulated in CL and CH groups, the expression of p-Akt and Bcl-2 was up-regulated, and the expression of Bax was down-regulated in group P , and the the expression of p-ERK1∕2 and c-fos was up-regulated in group M ( P<0. 05) . Compared with group CH, the MWT was significantly decreased, and the TWL was shortened in P and M groups, the expression of p-ERK1∕2 and c-fos was down-regulated in group P, and the expression of p-Akt and Bcl-2 was down-regulated, and the expression of Bax was up-reg-ulated in group M ( P<0. 05) . Conclusion Curcumin can reduce lidocaine-induced nerve injury, and the mechanism may be partly related to enhancing the activity of Akt and ERK1∕2 signaling pathway in rats.

OBJECTIVE: To explore the genetic etiology for a child with profound intellectual disabilities and obvious behavioral abnormalities. METHODS: A male child who had presented at the Zhongnan Hospital of Wuhan University on December 2, 2020 was selected as the study subject. Peripheral blood samples of the child and his parents were collected and subjected to whole exome sequencing (WES). Candidate variant was verified by Sanger sequencing. Short tandem repeat (STR) analysis was carried out to determine its parental origin. The splicing variant was also validated in vitro with a minigene assay. RESULTS: WES results revealed that the child had harbored a novel splicing variant of c.176-2A>G in the PAK3 gene, which was inherited from his mother. The results of minigene assay have confirmed aberrant splicing of exon 2. According to the guidelines from the American College of Medical Genetics and Genomics, it was classified as a pathogenic variant (PVS1+PM2_Supporting+PP3). CONCLUSION The novel splicing variant c.176-2A>G of the PAK3 gene probably underlay the disorder in this child. Above finding has expanded the variation spectrum of the PAK3 gene and provided a basis for genetic counseling and prenatal diagnosis for this family.
Child ; Female ; Humans ; Male ; Pregnancy ; Exons ; Intellectual Disability/genetics* ; Mothers ; Mutation ; p21-Activated Kinases/genetics* ; Parents ; RNA Splicing

Objective:To explore the genetic etiology of a child with primary hypertrophic osteoarthropathy(PHO).Methods:A child who was admitted to Zhongnan Hospital of Wuhan University on July 27, 2021 was selected as the study subject. Genomic DNA was extracted from peripheral blood samples of the child and his parents and subjected to whole exome sequencing. Suspected splicing variant was verified by Sanger sequencing of family members. In vitro function was validated through a minigene assay, whilst the suspected exonic deletion was validated by long-fragment PCR. This study was approved by the Children′s Hospital Affiliated to Zhengzhou University (Ethics No. 2023-K-011). Results:Whole exome sequencing revealed that the child has harbored compound heterozygous variants of HPGD gene, including a heterozygous deletion (Exon 3 del) derived from his father and a splicing variant (c.421+ 1G>T) derived from his mother. Long-fragment PCR verified that the child and his father had both harbored a 7 565 bp heterozygous deletion (c.218-1304_324+ 6156del), whilst the minigene assay proved that the splicing variant has resulted in skipping of exon 4. Conclusion:The heterozygous c. 218-1304_324+ 6156del deletion and the c. 421+ 1G>T splicing variant of the HPGD gene probably underlay the pathogenesis in this child. Above finding has enriched the mutational spectrum of the HPGD gene and provided a basis for genetic counseling and prenatal diagnosis for this family.

Objective:To seek the optimal melanin-removal method for hematoxylin and eosin (HE) staining, immunohistochemistry and molecular detection.Methods:Thirty-eight paraffin tissue samples of malignant melanoma diagnosed at the Fujian Cancer Hospital, Fuzhou, China between January 2018 and March 2022 were collected and used to make a tissue microarray. Melanin in these cases was removed using warm hydrogen peroxide, double oxidation depigmentation, modified potassium permanganate-oxalic acid or trichloroisocyanuric acid, followed by HE staining. The cases were divided into two cohorts: one was subject to the one of the above four methods to remove melanin first, followed by immunohistochemistry (SOX-10, Ki-67, HMB45 and Melan A), while the other was subject to immunohistochemical staining first and then a melanin removal. Following that, seventeen melanin-rich paraffin tissue samples were collected and depigmented using the methods described above. DNA extraction was then done, followed by assessments of DNA content and quality. Moreover, the completeness of melanin removal, the effect on HE and immunohistochemical staining, and the quality of DNA were compared between the depigmented methods.Results:Regarding the effectiveness of melanin removal, the modified potassium permanganate-oxalic acid and the warm hydrogen peroxide methods were the most effective, and both showed residual melanin in only 5.26% (2/38) of the cases. The trichloroisocyanuric acid method showed residual melanin in 10.53% (4/38) of the cases. The worst was the double oxidation depigmentation method, which showed pigment residue in 15.79% (6/38) of the cases. For HE staining, the percentage of good staining with the warm hydrogen peroxide method was 92.11%, higher than the other three methods. For immunohistochemical staining, the mean staining scores of immunohistochemistry first followed by melanin removal with modified potassium permanganate-oxalic acid, double oxidation and trichloroisocyanuric acid were 20.84, 26.63 and 35.02, respectively. These immunohistochemical staining scores were higher than those of melanin removal first followed by immunohistochemistry (8.70, 15.41 and 21.22, respectively). The mean staining score of melanin removal by warm hydrogen peroxide method followed by immunohistochemistry was 33.57, superior to that of immunohistochemistry followed by the melanin removal (19.96). Moreover, the staining scores of HMB45, MelanA and Ki-67 with immunohistochemical staining followed by trichloroisocyanuric acid method were 36.45, 33.79, and 36.24, respectively, while the staining score of SOX10 with melanin removal by warm hydrogen peroxide followed by immunohistochemistry was 34.39. The DNA was significantly degraded by modified potassium permanganate-oxalic acid, double oxidation depigmentation and trichloroisocyanuric acid, whereas the mean concentration of DNA extracted after melanin removal by hydrogen peroxide method was 59.59 μg/L, substantially higher than that of DNA extracted without melanin removal (30.3 μg/L, P=0.001). The A260/ A280 of DNA extracted after melanin removal by hydrogen peroxide was between 1.8 and 2.0 in all cases, and the A260/ A230 was above 2.0 in sixteen cases, suggesting high purity of DNA. However, the DNA extracted without removing the melanin showed poor purity, with A260/ A280 below 1.8 in eight cases and A260/ A230 below 2.0 in sixteen cases. Conclusions:Warm hydrogen peroxide showed the least melanin residue, superior HE staining and a minimal effect on DNA purity/quality compared to the other three methods. It thus appears most suitable for PCR, NGS and other molecular detection. Melanin removal with trichloroisocyanuric acid after immunohistochemical staining has the least melanin residual, and thus could be the most convenient and efficient. However, it is noted that the efficacy of the same depigmentation method varies with different antibodies. Therefore, the optimal depigmentation method should be selected based on the specific markers of interest.

Objective:To explore the effects of attribution training on coping style, stigma, quality of life and negative emotions of stroke patients.Methods:From June 2019 to June 2020, 85 patients with ischemic stroke admitted to the First Affiliated Hospital of Zhengzhou University were selected by simple random sampling. The patients were randomly divided into the control group (42 cases) and the observation group (43 cases) . The control group was given routine nursing, while the observation group received attribution training on the basis of the control group. Before and one and three months after the intervention, the patients in the two groups were evaluated with the Attributional Style Questionnaire (ASQ) , the Stigma Scale for Chronic Illness-8 (SSCI-8) , the World Health Organization Quality of Life-100 (WHOQOL-100) , the Self-Rating Anxiety Scale (SAS) , and the Self-Rating Depression Scale (SDS) .Results:There were statistical differences between the two groups in time main effect, intervention main effect, interaction between intervention and time of ASQ, SSCI-8, WHOQOL-100 scores ( P<0.05) . Before intervention, there was no statistical difference in SAS and SDS scores between the two groups ( P>0.05) . Three months after the intervention, the SAS and SDS scores of the two groups were lower than those before the intervention, and those of the observation group were lower than those of the control group, and the differences were statistically significant ( P<0.05) . Conclusions:Attribution training can effectively improve the coping style of stroke patients, improve their quality of life, reduce patients' stigma, anxiety and depression, which is worthy of clinical promotion.

Objective:To establish a scientifically standardized nursing quality evaluation index system that conformed to the characteristics of enteral nutrition in critically ill patients, with the aim of providing reference for evaluating the nursing quality of enteral nutrition in critically ill patients.Methods:Using the three-dimensional quality evaluation model of "structure-process-outcome" as the theoretical framework, the content of each level of the evaluation index system was preliminarily formulated by literature analysis. From May to July 2022, the Delphi expert consultation method was used to conduct two rounds of consultation with 21 experts to determine the index system.Results:In the two rounds of consultation, the effective response rates of the questionnaire were 85.71% (18/21) and 100.00% (18/18) , respectively. The expert authority coefficients were 0.867 and 0.878, respectively. The expert Kendall's W of the overall index were respectively 0.092 and 0.187 ( P<0.01) . The final nursing quality evaluation index system of enteral nutrition in critically ill patients included 3 first level indicators, 9 second level indicators, and 60 third level indicators. Conclusions:The constructed nursing quality evaluation index system of enteral nutrition in critically ill patients has put forward clear requirements and quality standards for enteral nutrition nursing for critically ill patients, which has good scientific and reliable characteristics and can provide reference for the nursing quality evaluation of enteral nutrition in critically ill patients.

Purpose To explore the clinicopathological features,diagnosis and differential diagnosis of large B-cell lym-phoma with IRF4 rearrangement(LBCL-IRF4r).Methods Clinical data of 8 cases of LBCL-IRF4r were collect,hematoxy-lin-eosin and immunohistochemical of EnVision two-step stains,in situ hybridization and FISH was used to study the histology,immunotypes and molecular genetic characteristics.The rele-vant literatures were reviewed.Results Among 8 cases of LBCL-IRF4r,the male to female ratio was 1.67:1,with age range 10-53 years(mean 25.8 years).Five cases occurred in tonsils,2 cases in nasopharynx and 1 cases in inguinal lymph node.Microscopically,the tumors presented with a purely follic-ular,purely diffuse or a combined follicular and diffuse architec-ture.The tumor cells were typical centroblasts and less frequent-ly medium-sized blastic cells with smaller nucleoli,apoptosis and nuclear fragmentation were easily seen.Immunophenotypi-cally,the tumor cells of the eight cases of LBCL-IRF4r diffuse strongly expressed CD20(8/8),PAX5(2/2),CD79a(3/3),BCL6(8/8)and MUM-1(8/8),mostly expressed CD10(7/8),partially expressed BCL2(5/8)and CD5(4/8),and did not express Cyclin D1,CD23 and CD30.The percentage of Ki67 index ranged from 70％to 95％.EBER in situ hybridiza-tion was negative in all cases.IRF4 rearrangements were detec-ted in all cases(8/8).BCL6 rearrangements were detected in one case(1/2).MYC(0/4)and BCL2(0/3)rearrangements were not detected in all cases.Conclusion LBCL-IRF4r is more common in children and adolescents with characteristic IG::IRF4 rearrangement and a good prognosis,which needs to be differentiated from other types of large B-cell lymphoma.

Patients with glioblastoma (GBM) generally have a bad prognosis and short overall survival after being treated with surgery, chemotherapy or radiotherapy due to the histological heterogeneity, strong invasive ability and rapid postoperative recurrence of GBM. The components of GBM cell-derived exosome (GBM-exo) can regulate the proliferation and migration of GBM cell via cytokines, miRNAs, DNA molecules and proteins, promote the angiogenesis via angiogenic proteins and non-coding RNAs, mediate tumor immune evasion by targeting immune checkpoints with regulatory factors, proteins and drugs, and reduce drug resistance of GBM cells through non-coding RNAs. GBM-exo is expected to be an important target for the personalized treatment of GBM and a marker for diagnosis and prognosis of this kind of disease. This review summarizes the preparation methods, biological characteristics, functions and molecular mechanisms of GBM-exo on cell proliferation, angiogenesis, immune evasion and drug resistance of GBM to facilitate developing new strategies for the diagnosis and treatment of GBM.
Humans ; Glioblastoma/genetics* ; Exosomes/metabolism* ; MicroRNAs/metabolism* ; Prognosis ; Cell Proliferation ; Brain Neoplasms/genetics* ; Cell Line, Tumor