Objective To constructan eukaryotic expression recombinant plasmid named pEGFP-N2-PRNP .Methods Total RNA was extracted from alzheimer (AD) disease peripheral blood ,and the PRNP gene was amplified by reverse transcription-poly-merase chain reaction(RT-PCR) .By using gene recombination technique ,human PRNP cDNA was inserted into retroviral vector pEGFP-N2 .The recombinant plasmid was identified by a pair of specified primers containing the restriction sites of Xho Ⅰ and EcoRⅠ .Results The PRNP gene could be obtained by RT-PCR ,the recombinant plasmid was identified by restriction endonucle-ase analysis ,PCR and sequence analysis ,and the expression vector pEGFP-N2-PRNP ,which could be stably expressed in SH-SY5Y cells .Conclusion The recombinant plasmid pEGFP-N2-PRNP is constructed successfully ,Which offers a basic for the further re-search on PRNP biological fuction .

Objective To investigate the clinic signification of newborn hearing screening combined with deafness susceptibility genes screening. Methods 1 440 newborns(3 ~ 5 days after birth) were screened for 8 hot spot hearing loss associated mutations from GJB2, mt12S rRNA and SLC26A4. At the same time, all infants received hearing screening. Those who failed to pass two-step test were referred to further audiological assessment. Results The carrier rate of commonmutations was 1.46% for GJB2 c.235delC, 0.35% for GJB2 c.299-300delAT, 0.42% for mt12S rRNA c.1555A > G, 0.42% for SLC26A4 c.IVS7-2A > G and 0.14% for SLC26A4 c.2168A > G. The total carrier rate was 2.78%. 10 infants were diagnosed as hearing loss in the hearing screening and follow-up audiology assessment (6.94‰) and 5 were diagnosed as severe hearing loss (3.47‰). 32 hearing loss associated mutation carriers passed the hearing screening. Conclusions Genetic screening of newborn hearing screening can be helpful to find out neonates with late-onset and progressive hearing impairment, which were significant for early intervention, regular follow-up and reduction of deafness.

Objective To explore the curative effects of unilateral Orthofix external fixation in the treatment of tibial defects.Methods From June 2012 to February 2016,we treated 19 patients with tibial defects using unilateral Orthofix external fixation.They were 14 men and 5 women,aged from 26 to 79 years (mean,43.0 years).The right tibia was involved in 12 cases and the left tibia in 7.The soft tissue defects averaged 6.4 cm × 3.7 cm.The mean bone defect before bone transport was 8.9 cm in length.The anteroposterior and lateral X-ray films of the affected tibia were taken regularly after surgery.Edwards tibial fracture scores were used to evaluate the functional recovery and complications of the affected limb at final follow-ups.Results All the patients were followed up for 10 to 40 months (average,18.5 months).Bone union time ranged from 4 to 16 months (average,9.0 months).Lengths of bone transport ranged from 6 to 10 cm (average,7.8 cm).All the bone defects were reconstructed.According to the Edwards scoring at the final follow-ups,10 cases were rated as excellent,6 as good and 3 as poor.Complications were observed in 4 cases,giving a complication rate of 21.1％.Conclusion Unilateral Orthofix external fixation and bone transport can lead to preferable curative effects in the management of tibial defects.

Objective: To explore the feasibility and clinical effect of single bundle anatomic intrathecal reconstruction of proximal injury of anterior cruciate ligament (ACL) using dissociate peroneus longus tendon combined with semitendinosus tendon. Methods: From January 2015 to September 2016, a total of 24 patients with proximal injury of ACL, confirmed by arthroscopy, were admitted to the sports medicine department of our hospital. The ACL was completely ruptured from the proximal footprint and the tibial side residual remained intact. There were 19 males and 5 females; 16 cases on the right side and 8 cases on the left side. The mean age was 27.88±7.13 years old; The interval between injury and surgery was 14.83±9.09 d; The dissociate peroneus longus tendon and semitendinosus tendonfrom the injured extremity were folded in half, then braided and trimmed into ACL graft for use. The ACL remnant was preserved and the graft was pulled through the stump for single bundle anatomic intrathecal reconstruction. The ACL graft was fixed with Endobutton on the femoral side and interference screw on the tibial side. The results of Lachman test, Lysholm scores, Tegner scores and International Knee Documentation Committee (IKDC) subjective scores of knee before operation and at final visit were recorded to evaluate the stability and function of the knee. Visual analogue score (VAS) was used to record the changes of pain at the site where the peroneal longus tendon was harvested. The American Orthopaedic Foot and Ankle Society (AOFAS) Ankle-Hindfoot Scores were recorded before injury and at the final visit to assess the effect of ankle function after peroneal longus tendon resection. Results: The diameter of the ACL graft made of dissociate peroneus longus tendon and semitendinosus tendon is 8.88±0.30 mm. All patients were followed up at the outpatient clinic, with an average of 34.38±5.40 months, and no serious complications such as rerupture and joint infection were found. There were 16 grade B, 6 grade C and 2 grade D for preoperative Lachman test, none with hard end point. At last vist, there were 23 cases of grade A and 1 grade B, all with hard end points for Lachman test. At the preoperative and final visit, the Lysholm scores of the knee joint were 35.20±11.92 and 94.29±2.92 (t=23.850, P=0.000). Tegner scores were 3.46±0.93 and 8.04±1.00 (t=16.653, P=0.000). The subjective IKDC scores of knee joint were 47.63±13.06 and 91.71±3.75 (t=15.972, P=0.000). At the final visit, all the indicators of the knee were improved compared with those before surgery, and the difference was statistically significant.The AOFAS scores before injury and at the final visit were 98.83 (2.78) and 98.17 (4.01), respectively, with no statistically significant difference (t=1.850, P=0.076) . The VAS scores for the harvest of theperoneus longus tendon were 4.50±1.41, 0.54±0.65 and 0.29±0.55 immediately and 6 months after the operation and at the last visit.There was statistically significant difference between the scores immediately and 6 months after the operation (t=14.900, P=0.001).There was statistically significant differences between the scores at 6 months after the operation and the final visit (t=2.770, P=0.011). Conclusion Single bundle anatomic intrathecal reconstruction of proximal injury of ACL using dissociate peroneus longus tendon and semitendinosus tendon is feasible with good clinical effect.

Purpose: The goal of this study was to describe the current status of oncology nurses' behaviors toward end of life (EOL) care in China and to explore the factors associated with oncology nurses’ behaviors toward EOL care. Methods: A cross-sectional design was applied and a convenience sample of 1038 oncology nurses from 22 grade A hospitals were recruited into this study. A general social demographic data questionnaire was administered, and the Chinese version of Nurses’ Behaviors of Caring for Dying Patients Scale was used to assess nurse behavior toward EOL care. The total score ranges from 40 to 200 points. Data were analyzed with SPSS 26.0 software. Results: Chinese oncology nurses' average score of holistic EOL care behaviors was 2.97 ± 0.59. Oncology nurses provide physical care most (3.81 ± 0.76), followed by family care (3.02 ± 0.86), and spiritual care (2.37 ± 0.67). Multiple regression analysis showed that a higher frequency of sharing EOL care experience with colleagues, in-service palliative care education, higher level of head nurse support for EOL patient care, more cases of EOL care, higher working position, and nurse's perceived high level of support were positively associated with behavior toward EOL care. These six factors explained 16.2% of the total variance. Conclusions The results may help provide a basis for converting behavior for EOL care among oncology nurses and design interventions to better improve quality of life for EOL patients with cancer in China.

Objective To detect the association between rs4646999 polymorphisms in the promoter region of the c-Jun and the prognosis of sporadic colorectal cancer. Methods rs4646999-673C>T genetypes were deter-mined by Taqman-MGB probes in 436 colorectal cancer cases. The survival curve was analyzed by Kaplan-Meier analysis and Cox regression.Western blot was used to analyze the expression levels of c-Jun protein in different gen-otypes. Results Univariate analysis showed that the cumulative survival rate of patients with rs4646999TT geno-type was significantly higher than that of patients with CT and CC genotype. Multivariate Cox regression analysis showed that the differentiation,lymph node metastasis,distant metastasis,TNM stage and rs4646999 genetypes were prognostic factors.Compared with the carriers of TT genotype,CT/CC complex genotypes were associated with poor prognosis of colorectal cancer(P<0.05).Protein expression analysis showed that the expression of c-Jun pro-tein in CC genotype was increased.In contrast,the TT genotype was decreased.Conclusions This study provided the evidence that rs4646999-673C>T genetic variant in c-Jun promoter regions is associated with the poor survival prognosis of colorectal cancer,possibly by elevating the protein expression levels that appeared to up-regulate activ-ity of c-Jun thus tumorigenesis.

Objective:To analyse the clinical presentation and pathogenic gene mutations of a family diagnosed with primary familial brain calcification (PFBC).Methods:A pedigree with primary familial brain calcification was recruited. The clinical data of the proband who was admitted to the Affiliated Hospital of Guizhou Medical University in March 2020 and the family members were collected. The DNA sequence of myogenesis regulating glycosidase (MYORG) gene was detected by Sanger sequencing in the proband and some available family members.Results:The proband is a male, 30 years old. There was only one patient of PFBC in this family. The first symptom of the proband was vagueness of speech, and gradually extrapyramidal symptoms such as slow and flexible movement and advanced cognitive impairment appeared. The brain CT of the proband and his second brother showed extensive symmetrical calcifications, mainly located in the bilateral cerebellar hemispheres, basal ganglia and thalamus. A homozygous mutation in the exon 2 of the MYORG gene [c.1967T>C(p.I656T)] was identified in the proband and an asymptomatic patient. The heterozygous mutation of MYORG gene was also detected in four healthy family members.Conclusions:All patients with homozygous mutations of MYORG gene showed calcification in CT scan, and most of the lesions were located in basal ganglia, cerebellum, subcortical white matter and thalamus. Compared with the patients with autosomal dominant gene mutation, the patients with MYORG gene mutation had more extensive intracranial calcification lesions, and the pontocerebellar lesions were more common. The most common symptoms of MYORG gene mutation patients were dyskinesia, mainly tremor paralysis and unclear speech.

The intestinal flora and gut barrier function are of great significance for gut function and human health. When the intestinal flora is disrupted and the gut barrier structure is disrupted, it can lead to bacterial translocation, endotoxin influx into the bloodstream, and the production of pro-inflammatory factors, leading to various tissue damage in the body. Tongfu method in TCM can affect the intestinal environment by regulating intestinal permeability and immune response, restoring normal intestinal movement, and regulating the structure and metabolites of intestinal flora, thereby maintaining intestinal homeostasis and body health. The research on regulating intestinal flora and improving intestinal barrier function by Tongfu method can provide reference for further research on the relationship between TCM and intestinal microecology, and provide ideas for clinical treatment.

Objective: To investigate the effect of Kamistad gel on oral ulcer healing and the expression of tumor necrosis factor α (TNF-α), interleukin 6 (IL-6) and epidermal growth factor (EGF) after oral administration in ulcer tissue of rats and to provide animal experimental data for the clinical application of Kamistad gel. Methods: The oral ulcer rat model was established by chemical cauterization. The rats were randomly divided into four groups: Kamistad group (Kamistad gel), befuxin group (befuxin gel), lidocaine group (lidocaine cream), blank control group (normal saline), with 12 rats in each group. The ulcer area of the rats in each group was measured before and 1, 3 and 5 days after treatment; at 1 day after treatment, the duration of swabbing behavior within 3 minutes of intraoral capsaicin infusion was recorded to evaluate the degree of pain; the ulcer tissue was collected at 5 days after treatment, and the histopathological changes were observed by HE staining, the expression of TNF-α, IL-6 and EGF in the ulcer tissue was detected by immunohistochemistry and ELISA. Results: At 1 day after treatment, the duration of mouth wiping induced by capsaicin was significantly shorter in the Kamistad group than in the blank control and befuxin groups (P < 0.05), but there was no significant difference between the Kamistad and lidocaine groups (P >0.05). At 5 days after treatment, the ulcer area was significantly smaller in the Kamistad group than in the blank control and lidocaine groups (P < 0.05), and there was no significant difference between the Kamistad and befuxin groups (P >0.05). At 5 days after treatment, H&E staining of the oral ulcer tissue sections showed significantly reduced levels of inflammatory cells and significantly proliferated fibroblasts and better epithelial hyperplasia in the Kamistad group compared with those in the lidocaine and blank control groups, and there were no differences between the Kamistad and befuxin groups. At 5 days after treatment, the expression levels of TNF-α, IL-6 and EGF in the ulcer tissue of rats in each group were significantly different (P < 0.05). Compared with the blank control and lidocaine groups, the expression of TNF-α and IL-6 was significantly decreased and the expression of EGF was significantly increased in the Kamistad group (P < 0.05); there were no significant differences in the expression of the above three factors between the Kamistad and befuxin groups (P > 0.05). Conclusion Kamistad gel exhibited anti-inflammatory, analgesic and healing effects on experimental oral ulcers.