Objective: To design the ribozymes to cleave human TIMP 1 mRNA, and embed them into U 6snRNA to make them stable. Methods: Ribozymes were designed according to the “hammerhead structure” described by Symons.Computer was used to analyze the possible cleavage sites. Results: Three ribozymes targeting the nt123, nt299 and nt353 on TIMP 1 mRNA were designed. Embedding ribozyme in U 6snRNA had little effect on its binding with the substrate. Conclusion: Computer assisted design is indispensable in studying ribozyme. Embedding ribozymes in U 6snRNA may be a good way to solve the problems existing in ribozyme study. [

Objective. The goal of this paper is to investigate the relationship between the N-glycosylation of acetylglucosaminyltransferase V(Glc NAcT-V) and its activity and to know which site among the 6 N-glycosylation sites in the GlcNAcT-V gene is the most important.Methods. Wild type of GlcNAcT-V was transfected into COS-7 cells and its activity was measured 48 h later. The first site (Asn 110) was mutated with site-directed mutagenesis and transfected into COS-7 cells.Results. It was found that after the cells were added tunicamycin(TM, 1 μ g/ml), the activity was 11.7% of the wild type. The activity of the cells with mutating GlcNAcT-V was about 12.0% of the wild type. RT-PCR showed that there was no significant change in mRNA expression among the three groups.Conclusion. The N-glycosylation is important for its activity. Our results suggest that the N-linked carbohydrates on GlcNAcT-V are required for the posttranscriptional activity of the enzyme.

Objective:To investigate the effects of hyperthermia on the biological behavior of human laryngeal cancer Hep-2 cisplatin-resistant (Hep-2/CDDP) cell line and its possible mechanism.Methods:Hep-2/CDDP cell line was induced by high impact combined with increasing concentration method. Cell count method was used to detect the cell proliferation ability of Hep-2 parental cell group (Hep-2 cells without cisplatin-resistance and the cells were cultured with RPMI 1640 cultured medium without cisplatin), Hep-2/CDDP cell group and Hep-2/CDDP+cisplatin group (using RPMI 1640 cultured medium including 4 mg/L cisplatin). Hep-2/CDDP cell group and Hep-2 parental cell group were treated with cultured medium including 0, 0.004, 0.04, 0.4, 4, 40 mg/L cisplatin, respectively. The sensitivity of Hep-2/CDDP cells to cisplatin, vincristine and 5-fluorouracil was determined by using methyl thiazolyl tetrazolium (MTT) method. The half inhibitory concentration ( IC50) and resistance index (RI) were also calculated. Hep-2/CDDP cell group was divided into 4 subgroups: the cells in the control group were cultured for 24 h at 37 ℃; the cells in hyperthermia group were treated at 43 ℃ for 2 h and then re-cultured at 37 ℃ for 22 h; the cells in cisplatin group were cultured at 37 ℃ for 24 h in cultured medium containing 4 mg/L cisplatin. The cells in hyperthermia combined with cisplatin group were cultured in cultured medium containing 4 mg/L cisplatin, treated at 43 ℃ for 2 h and then re-cultured at 37 ℃ for 22 h. The effects of hyperthermia combined with cisplatin on the proliferation and early apoptosis of Hep-2/CDDP cells were detected by using MTT and flow cytometry. The interaction of hyperthermia combined with cisplatin on the proliferation and early apoptosis of HEP-2/CDDP cells was observed by using factorial analysis. Western blotting was used to detect the effect of hyperthermia combined with cisplatin on the expressions of wild-type p53 and PI3K in Hep-2/CDDP cells. Hep-2/CDDP cells were divided into 4 groups: the control group (Hep-2/CDDP cells were cultured for 24 h at 37 ℃); chemotherapy group was treated with 12 mg/L vincristine or 9 mg/L 5-fluorouracil; in the hyperthermia group, Hep-2/CDDP cells were treated at 43℃ for 2 h and then re-cultured at 37 ℃ for 22 h; in hyperthermia combined with chemotherapy group, the cells were cultured in a medium containing 12 mg/L vincristine or 9 mg/L 5-fluorouracil, treated at 43 ℃ for 2 h and then re-cultured at 37 ℃ for 22 h. MTT method was used to detect the effect of hyperthermia combined with vincristine and 5-fluorouracil on the proliferation of Hep-2/CDDP cells. Results:Hep-2/CDDP cell line was successfully established. There were no significant differences in the number of cells in Hep-2/CDDP cell group, Hep-2 parental cell line group and Hep-2/CDDP + cisplatin cell group at different time points (all P > 0.05), and the doubling time was 43.8, 40.6 and 43.5 h, respectively. The IC50 of Hep-2 parental cell line group and Hep-2/CDDP cell group to cisplatin was 4.771 mg/L and 42.749 mg/L, respectively, and the RI was 8.960. Hyperthermia combined with cisplatin could inhibit the proliferation of Hep-2/CDDP cells ( F = 327.91, P < 0.05) and promote the early apoptosis of Hep-2/CDDP cells ( F = 724.63, P < 0.05). Factorial analysis showed that hyperthermia combined with cisplatin had an interaction effect on the proliferation and early apoptosis of Hep-2/CDDP cells ( F = 185.68, 472.51, all P < 0.05). Western blotting showed that the relative expression levels of wild-type p53 protein and PI3K protein in the control group, hyperthermia group, cisplatin group and hyperthermia combined with cisplatin group were significantly different ( F = 547.76, 404.44, all P < 0.01). Hyperthermia combined with vincristine or 5-fluorouracil could inhibit the proliferation of Hep-2/CDDP cells ( F = 33.06, 34.61, all P < 0.05). Factorial analysis showed that hyperthermia combined with vincristine and 5-fluorouracil had no interaction effect on the proliferation of Hep-2/CDDP cells ( F = 0.64,0.60, all P > 0.05). Conclusions:Hyperthermia may reverse the resistance of Hep-2/CDDP cell line to cisplatin by upregulating wild-type p53 expression and inhibiting the PI3K pathway. Hep-2/CDDP cell line has cross-resistance to vincristine and 5-fluorouracil. Hyperthermia can increase the sensitivity of Hep-2/CDDP cell line to vincristine and 5-fluorouracil.

OBJECTIVE To discuss the relationship between the main concentration of air-borne pollen and the number of visits for allergic diseases in Chengde City. METHODS The daily airborne pollen concentration and species in Chengde City,Hebei Province were monitored by gravity sedimentation method from March 1 to October 15,in 2020-2022. The number of visits of patients with allergic rhinitis,asthma and conjunctivitis in department of otorhinolaryngology,respiratory and ophthalmology were counted by HIS system in the hospital computer center,furthermore,the correlation between pollen concentration and the number of visits of the above diagnosed patients was analyzed. RESULTS The pollen concentration in Chengde showed two peaks each year,which were in May in spring and August in autumn. The main pollen sources in spring were Pinaceae,ash,birch,ulmaceae,Cupressaceae,Moraceae and Rosaceaceae,accounting for 82.6％ of the total annual pollen(78.2％,83.5％,and 86.1％ in 2020-2022,respectively). In autumn,artemisia and humulus were the main sources of pollen,accounting for 9.94％ of the total annual pollen(11.4％,6.3％,12.11％,in 2020-2022,respectively). The number of outpatient visits patients with allergic rhinitis,asthma and conjunctivitis were higher in May and August than in other months,and the number of patients with allergic rhinitis in August was significantly increased compared with other months(P<0.05). The total monthly visits of the three departments and the visits of the three allergic diseases were positively correlated with pollen concentration. CONCLUSION The pollen concentration in the spring and autumn seasons of Chengde City has a direct impact on allergic diseases. Monitoring and broadcasting pollen concentration can provide early warning for doctors and patients,and provide effective information for early intervention,protection,and reasonable treatment of diseases.

Objective:To evaluate the association between the eye exercises and one-year axial eye elongation in grade 7 students in Beijing.Methods:Sampling was performed using a multistage random cluster approach, and 1 443 students of grade 7 were selected from 9 middle schools in 6 districts for the baseline survey. Data were collected by questionnaires and axial length measurement. Multiple linear regression analysis was used to evaluate the association between eye exercises and excessive axial eye elongation.Results:Among 1 197 (82.95%) students with complete information, the median ( Q R) age was 12.00 (1.00) years old, girls accounted for 44.28%, and the median ( Q R) axial eye elongation was 0.22 (0.18) mm. In the multiple linear regression analysis, the frequency of eye exercises was significantly correlated with excessive axial eye elongation in boys ( β=-0.135, 95% CI:-0.253--0.018) but not in girls ( β=-0.075, 95% CI:-0.207- 0.058) after adjusting for sex, age, body height, the number of myopic parents, time spent outdoors and time spent on reading and writing outside class; while the seriousness of eye exercises was not significantly associated with axial eye elongation in boys ( β=-0.028, 95% CI: -0.114-0.058) and girls ( β=-0.035, 95% CI: -0.134-0.064). Conclusion:The increased frequency of eye exercises is beneficial to control the axial eye elongation in boys in Beijing.

ObjectiveTo explore the characteristics of balance and cortical activation in older adults when performing cognition-balance dual tasks. MethodsFrom January to April, 2023, 20 healthy older adults were non-targeted recruited. They completed six tasks of close eyes & fixed platform (CF), close eyes & fixed platform & cognitive task (CFc), open eyes & sway-referenced platform (OS), open eyes & sway-referenced platform & cognitive task (OSc), close eyes & sway-referenced platform (CS), and close eyes & sway-referenced platform & cognitive task (CSc) on the Balance SD, wearing functional near-infrared spectroscopy caps. The overal stability index (OSI) was measured with Balance SD. The premotor cortex (PMC), sensorimotor cortex (SMC) and prefrontal cortex (PFC) were as regions of interest (ROIs), and the β values were calculated. ResultsThe OSI was more as CFc than as CF (Z = -2.014, P < 0.05), and was less as CSc than as CS (Z = -2.063, P < 0.05). The β values of bilateral ROIs were all more as CFc than as CF (|Z| > 2.464, |t| > 3.733, P < 0.05), and as OSc than as OS (|t| > 2.308, P < 0.05); the β value of the right SMC was more as CSc than as CS (t = -2.912, P < 0.05). The number of correct counts was less as CSc than as CFc and OSc (|Z| > 3.643, P < 0.001). ConclusionBalance has been impaired under dual tasks for older adults, while activation of cerebral cortex increases. However, for more difficult balance task, older adults would preferentially maintain postural balance under dual tasks, while cognitive performance decreases, which may be the results from no more activation of cerebral cortex under dual tasks.

Objective: To clarify the situation of the detection of upper gastrointestinal cancer and precancerous lesions in the Feicheng city and discuss the possible influencing factors. Methods: A cluster sampling method was used to determine the participants. A unified questionnaire was used to investigate the basic information including history of alcohol intake, smoking and chinese tea, as well as other eating habits, medical history of digestive tract and cancer. Endoscopy was used to to screen the patients with upper gastrointestinal cancer and precancerous lesions. Influential factors were explored by non-conditional logistic regression model. Results: 911 of 7 291participants were positive, and the total detection rate was 12.49%. The total positive detection rate of male and female was 17.94% and 8.71%, respectively (P<0.001). Multivariate logistic regression analysis showed that the sex (OR=0.527, 95%CI: 0.440-0.631), age (OR=2.037, 95%CI: 1.849-2.245), smoking (OR=1.240, 95%CI: 1.014-1.516) and alcohol consumption (OR=1.232, 95%CI: 1.012-1.500) , meat and protein intake (OR=0.794, 95%CI: 0.638-0.987) and drink tea (OR=1.233, 95%CI: 1.056-1.440) may be influencing factors of the total detection rate of upper gastrointestinal cancer and precancerosis. Conclusions In Feicheng city, intake of meat and protein is the protective factor of the upper gastrointestinal cancer and precancerous lesions. Men, aged, smoking, drinking and tea will increase the risk of upper gastrointestinal cancer and precancerous lesions.

ObjectiveTo explore the possible mechanisms of Tongfengning （痛风宁， TFN） in treating hyperuricemia （HUA） of spleen deficiency with exuberance of dampness syndrome. MethodsTen of 60 mice were randomly selected， and were fed with regular diet as the control group， while the remaining 50 mice were fed with high-fat and high-sugar diet combined with excessive exercise and potassium oxonate-allopurinol suspension to establish an HUA animal model of syndrome of spleen deficiency with exuberance of dampness. After the successful modeling， in order to better observe the effects of TFN on the intestinal microbiota of the model mice， a mixed antibiotic suspension was administered by gavage to induce further dysbiosis of the intestinal microbiota in the model mice. Fifty sucessfully modeled mice were randomly divided into model group， TFN group， allopurinol group， probiotics group， and an allopurinol + probiotics group， 10 in each group. The TFN group was administered TFN liquid at a dosage of 19.11 g/（kg·d） by gavage. The allopurinol group was administered allopurinol suspension at a dosage of 78 mg/（kg·d） by gavage. The probiotics group was administered live combined Bifidobacterium and Lactobacillus tablets suspension at a dosage of 3 g/（kg·d） by gavage. The allopurinol + probiotics group was administered allopurinol at a dosage of 78 mg/（kg·d） and live combined Bifidobacterium and Lactobacillus tablets suspension at a dosage of 3 g/（kg·d） by gavage. The control group and model group were administered normal saline at a dosage of 19.11 ml/（kg·d） by gavage. The interventions were continued for 21 days. In order to maintain a stable high blood uric acid state， all groups but the control group continued modeling while receiving drug intervention. The changes in spleen deficiency syndrome scores， blood uric acid levels， microbial community structure， acetic acid and butyric acid content in intestinal lavage fluid， adenosine deaminase （ADA） and xanthine oxidase （XOD） content in small intestine tissue， as well as ATP-binding cassette transporter G2 （ABCG2）， glucose transporter 9 （GLUT9） protein and mRNA expression in the small intestine tissue were compared among the groups of mice. ResultsCompared with the control group， the model group showed increased spleen deficiency syndrome scores， blood uric acid levels， relative abundance of phylum Firmicutes， Firmicutes/Bacteroidetes ratio， abundance of Bacteroides genus， Klebsiella genus， and Enterococcus genus， acetic acid content in intestinal lavage fluid， ADA and XOD content in small intestine tissue， as well as GLUT9 protein and mRNA expression （P<0.05）. The number of operational taxonomic units （OTUs） of intestinal microbiota， relative abundance of Bacteroidetes phylum， abundance of Lactobacillus genus and uncultured Bacteroides genus， butyric acid content in intestinal lavage fluid， and ABCG2 protein and mRNA expression in small intestine tissue were significantly decreased （P＜0.05）. Compared with the model group， in the group treated with TFN， probiotics， and allopurinol + probiotics， the spleen deficiency syndrome score， blood uric acid level， relative abundance of Firmicutes， acetic acid content in intestinal lavage fluid， ADA and XOD content in small intestine tissue， GLUT9 protein and mRNA expression significantly decreased. The number of gut microbiota OTUs， relative abundance of proteobacteria， butyric acid content in intestinal lavage fluid， ABCG2 protein and mRNA expression in small intestine tissue significantly increased （P＜0.05）. In the probiotics group， the ratio of Firmicutes to Bacteroidetes decreased. In the TFN group， the abundance of Lactobacillus and uncultured Bacteroidetes significantly increased， while the abundance of Parabacteroides， Klebsiella， and Enterococcus significantly decreased （P＜0.05）. Compared with the TFN group， allopurinol group and the probiotics group showed elevated blood uric acid levels， abundance of Bacteroidetes， ADA and XOD levels in intestinal tissue， and GLUT9 mRNA expression. The relative abundance of Firmicutes， abundance of lactobacilli， and ABCG2 mRNA expression significantly decreased. The probiotics group showed elevated GLUT9 protein expression in intestinal tissue. The probiotics group and the allopurinol plus probiotics group showed significantly higher scores for spleen deficiency syndrome in mice， and lower levels of butyric acid in mouse intestinal lavage fluid. The allopurinol group showed decreased numbers of OTUs in mouse intestinal flora， decreased abundance of proteobacteria， and butyric acid levels in intestinal lavage fluid. The allopurinol group also showed decreased ABCG2 protein expression in intestinal tissue， increased acetic acid levels in intestinal lavage fluid， increased abundance of Klebsiella， and significantly elevated GLUT9 protein expression （P＜0.05）. ConclusionsThe treatment of HUA with TFN may be associated with the regulation of intestinal probiotics （such as lactobacilli） and pathogenic bacteria （such as Klebsiella）， as well as the production of bacterial metabolites such as acetic acid and butyric acid. It may also involve reducing the expression of ADA and XOD in the intestines， decreasing intestinal uric acid production， upregulating the expression of intestinal epithelial urate transporter ABCG2， downregulating GLUT9 expression， and promoting intestinal uric acid excretion. These factors are related to the syndrome of spleen deficiency with exuberance of dampness.