Objective To investigate the effect of neuropeptide Y (NPY) on the osteoblastic differentiation of murine MC3T3-E1 cells and its mechanism related to the Wnt signaling pathway.Methods The murine MC3T3-E1 cells were divided into 4 groups according to the stimulators added:phosphate buffered saline (PBS) (control) and different concentrations of NPY (10-8 mol/L,10-10 mol/L and 10-12 mol/L).The cellular proliferation was detected with MTT assay after 1,3,5,7 and 9 days.The cells were identified with cell immunochemistry and Western Blot to find out the most effective concentration of NPY at different time points under osteoblastic condition.The cells were then divided into 4 groups:PBS,NPY,NPY + NPY receptor antagonist,and NPY + DKK1.Western blot was used to determine the expression of β3-catenin and p-GSK-3β in each group.Nuclear signaling activity of β3-catenin was observed using immunofluorescence staining.Results NPY significantly improved the proliferation of MC3T3-E1 cells at 7 and 9 days (P ＜0.05).NPY (10s mol/L and 10-10 mol/L) groups and NPY (10-10 mol/L and 10-12 mol/L) groups significantly improved the ALP activity at 4 and 14 days respectively (P ＜ 0.05).At 4 days,the expression of ALP protein was significantly decreased in the NPY + DKK1 group and the NPY + NPY receptor antagonist group compared with that in the NPY group (P ＜ 0.05).Although the expression levels of [β-catenin and p-GSK-3β protein were uninfluenced in either case,NPY significantly stimulated the nuclear signaling activity of β3-catenin.Conclusions NPY may significantly increase the expression of ALP protein in MC3T3-E1 ceils during osteoblastic differentiation.This effect might be mediated through the canonical Wnt signaling pathway.

Objective:To map dominant antigenic determinants on SAK by gene-targeted fragmemt library . Methods:①The PcAbs specificly against SAK were produced by immunning BALB/C mice. Pu rified the antisera through a SAK-Sepharose 4B affinity chromatography column. The purified PcAbs were biotinylated for next step. ② After constructing SAK r andom epitope library we sequenced 12 isolated clones randomly to ensure its int egrity ,capability and randomness.③The library was screening by situ-clone hy bridization.④Constructed mSAK ,the A1 region deleted mutant of SAK ,and used Western-blot assay to identified its immunoreactivity. Results:①Got a dominant epitope at amino acid 71-89,called A1 region; ②Western-bl ot assay suggested that mSAK, a mutant SAK without A1 region., didn't combine to the anti-SAK PcAbs.Conclusion:A dominant epitope of SAK was mapped successfully with a simple,effective method . [

Objective To investigate the characteristics of blood donors’psychological activity,take reasonable intervention measures to improve the success rate of blood donation recruitment and the ratio of repeated blood donation.Methods The data of blood donors’psychological activity was collected by distributing questionnaires randomly,and the psychological characteristics and worries were analysed.Results In terms of the blood donation purpose,there were 62.73% of the blood donors who donate blood for the first time and take the“utility psychological”as the principal thing.There were 76.01% of the blood donors who donate blood repeatedly and take the“dedication psychological”as the principal thing.In terms of wor-ries,there was 72.69% of the blood donors who donate blood for the first time and take the“safety of blood donation”as the principal thing.There was 77.91% of the blood donors who donate blood repeatedly and take the “service quality of blood donation”as the principal thing.Conclusion The success rate of blood donation recruitment and the ratio of repeated blood donation could be effectively improved by attaching importance to the psychological needs and worries of blood do-nors,by taking different psychological intervention measures strategies for different kinds of blood donors,and by meeting their needs and eliminating their worries.

Objective: To investigate the creation of smoke-free environments in public places in Hangzhou City, so as to provide insights into effective implementation of the tobacco control policy. Methods: The party and government administrations at each level, medical institutions, educational places, restaurants and entertainment places, and open public places were enrolled. The creation of smoke-free environments was investigated in these places through undercover investigation with field observations and concealed photography by a third-party professional investigation company from November to December, 2022. The building of smoke-free environments (totally 60 scores) and no smoking indoors (totally 40 scores) were evaluated according to the Criteria for Scoring of Smoke-free Organizations in Hangzhou City. Results: Totally 909 places were investigated, and the comprehensive score of smoke-free environment building was (82.83±14.13) points. There were 285 party and government administrations with a comprehensive score of (84.19±12.85) points, 65 medical institutions with a comprehensive score of (90.35±6.95) points, 65 educational places with a comprehensive score of (83.43±16.81) points, 403 dining and entertainment places with a comprehensive score of (80.68±14.75) points, and 91 open public places, with a comprehensive score of (82.34±14.77) points. There were 397 places with standardized tobacco control tips at entrances (43.67%), 308 places with tobacco control signs posted as required (33.88%), 707 places that set outdoor smoking areas correctly (77.78%), 68 places with smoking paraphernalia (7.48%), 28 places with tobacco sales (3.08%). There were 732 places without signs of indoor smoking (80.53%), 850 places without indoor smoking (93.51%) and 24 places without dissuading from smoking (2.64%). Conclusion The indoor no-smoking is overall satisfactory in public places in Hangzhou City; however, standardizing no-smoking tips at entrances, standardizing the posting of no-smoking signs and assignment of tobacco control materials remain to be improved.

Objective:To explore the inhibitory mechanism of herb cake-partitioned moxibustion on tumor growth in colitis-associated colorectal cancer(CAC)based on histone lysine demethylase 4D(KDM4D). Methods:Inbred male Sprague-Dawley rats were randomly divided into a normal group,a CAC group,a herb cake-partitioned moxibustion group,and an inhibitor group.Except the normal group,rats in the other three groups were treated with azoxymethane(AOM)combined with dextran sulfate sodium(DSS)to make CAC rat models.Rats in the normal group and the CAC group did not receive interventions;rats in the herb cake-partitioned moxibustion group received moxibustion at Qihai(CV6)and bilateral Tianshu(ST25),2 cones for one point each time,once a day for 30 d with 1-day rest every week;rats in the inhibitor group received intraperitoneal injection of KDM4D inhibitor,5-chloro-8-hydroxyquinoline(5-c-8HQ),once a day for 30 d.After intervention,the general condition,colon length,tumor number and volume,and histopathological colon changes were observed.The expression of adenomatous polyposis coli(APC),axis inhibitor(Axin),cyclin D1,matrix metalloproteinase(MMP)-7 and MMP-9 mRNAs were detected by real-time quantitative polymerase chain reaction.The proliferating cell nuclear antigen(PCNA),cleaved caspase3,KDM4D,APC,and Axin proteins were detected by immunohistochemistry. Results:Compared with the normal group,the general condition was poor,the colon length was significantly shortened(P<0.01),the number and volume of colonic tumors were increased(P<0.01),the structure of glandular duct was obviously disordered with"back-to-back"and cowall phenomenon,and also high-grade adenocarcinoma formed;the protein expression levels of PCNA and KDM4D were significantly increased(P<0.01),while cleaved caspase3,APC,and Axin were significantly reduced(P<0.01);the mRNA expression levels of cyclin D1,MMP-7,and MMP-9 were significantly increased(P<0.01),while APC and Axin were significantly reduced(P<0.01)in the CAC group.Compared with the CAC group,the general condition was improved,the length of colon was significantly increased(P<0.01),the number and volume of the colonic tumors were reduced(P<0.05),and the colon tissues showed epithelial cell proliferation with enlarged and deep staining nuclei,dysplasia and inflammatory cell infiltration;the protein expression levels of PCNA and KDM4D were significantly reduced(P<0.01),while the cleaved caspase3,APC,and Axin were significantly increased(P<0.01);the mRNA expression levels of cyclin D1,MMP-7,and MMP-9 were reduced(P<0.05),while the APC and Axin were increased(P<0.05)in the colon tissues of rats in the herb cake-partitioned moxibustion group and the inhibitor group. Conclusion:Herb cake-partitioned moxibustion regulated abnormally expressed KDM4D in CAC rats,activated APC and Axin,the upstream molecules of Wnt/β-catenin pathway,inhibited abnormally activated downstream molecules of Wnt/β-catenin pathway.This may be a key mechanism of herb cake-partitioned moxibustion in inhibiting CAC tumor growth.

Objective To establish a three-dimensional finite element model of pelvic anteroposterior compression (APC) for analysis of mechanisms for related liganentous damages.Methods A finite element model and a laboratory mechanical model of APC were established using the same pelvic specimens.In a finite element model of normal pelvic bones and ligaments,after the right pelvis was fixated the pubic symphysis (PS) was sectioned.Next,a manual external mobile force was gradually applied to the left hemipelvis to make the PS diastasis 10,20,30,40,60,80 and 100 mm apart.The mechanical experiment revealed the anterior sacroiliac ligament (ASIL) was ruptured when the PS diastasis reached 28 mm.After the strain value of ASIL was calculated through the finite element model,it was applied to the other pelvic ligaments.The displacement in front of the sacroiliac joint (SIJ),stress,strain and extent of injury and disruption of sacrotuberous/sacrospinous ligaments (STL/SSL) with a corresponding PS diastasis were observed and recorded.Results ASIL failed at the point when the PS diastasis was 28 mm and the displacement in front of SIJ was 7.41 ± 1.14 mm.The strain and maximum principal stress of ASIL calculated in the finite element model were 259.5％ and 543.24 MPa respectively.The maximum principal stress value of SSL was 35.00 MPa at the point of failure when the PS diastasis and the displacement in front of SIJ were 51 mm and 15.23 ±2.88 mm,respectively.When the PS diastasis and the displacement in front of SIJ were 100 mm and 7.5 mm respectively,the maximum principal stress value of STL was 16.17 MPa but the strained ligament was not ruptured.When the pelvis was rotated externally step by step,the ASIL failure was followed by the rupture of SSL but not necessarily by the STL failure.Conclusion As the finite element pelvic bone-ligament model established in this study can effectively simulate the mechanisms for APC injury,it can be used to evaluate different extents of pelvic ligamentous injury,providing a basis for the biomechanical study of pelvic bones and ligaments.

Objective To investigate the reliability of using the pubic symphysis diastasis of 25 mm and anterior separation distance of sacroiliac joint to differentiate anteroposterior compression (APC) type Ⅰ and Ⅱ injuries as well as assess the injury severity.Methods A total of 11 (seven males and four females) fresh cadaver specimens with 22 hemipelvis were collected.The pelvic APC injury test models including fixed hemipelvis (restricted group) and unfixed hemipelvis (non-restricted group) were established,with 11 hemipelvis in each group according to the random number table method.Meanwhile the specimens were divided into male group (14 hemipelvis) and female group (eight hemipelvis),simulating APC type injury external rotation hemipelvis.The public symophysis interval and anterior interval of sacroiliac joint of the original pelvis,the pubic symphysis diastasis and anterior diastasis of sacroiliac joint after anterior tibiofibular ligament failure,as well as the affected pelvis ligament and sacral ligament injury were recorded and compared between the restricted and non-restricted groups,male and female groups.Results There were no significant differences in the public symphysis interval of the original pelvis and anterior interval of sacroiliac joint between the restricted group and the non-restricted group (P ＞ 0.05).The pubic symphysis interval of the original pelvis was [(5.13 ± 0.61) mm] in male group and (4.03 ± 0.84)mm] in female group (P ＜ 0.05).When the anterior tibiofibular ligament ruptured,the pubic symphysis diastasis distance was (23.36 ± 7.27) mm,ranging from 12 to 41 mm,and the diastasis distance of anterior sacroiliac joint was (9.82 ± 3.25)mm,ranging from 5 to 18 mm.In terms of the public symphysis interval,there were no significant differences between male and female groups,restricted and the non-restricted groups (P ＞ 0.05).In terms of anterior interval of sacroiliac joint,there was significant difference between male and female groups (P ＜ 0.05) but no significant difference between the restricted and non-restricted groups (P ＞ 0.05).In the restricted group,sacrotuberous ligament injuries were found in four patients,and sacrospinous ligament injuries in five,whhile there were no obvious sacrospinous ligament and sacrotuberous ligament injuries in non-restricted group.There were 10 specimens with the pubic symphysis diastasis ≥23.36 mm and 10 specimens with the diastasis distance of anterior sacroiliac joint ≥9.82 mm (46％),and there were 15 specimens with at least the pubic symphysis interval ≥ 23.36 mm or the anterior interval of sacroiliac joint ≥ 9.82 mm (68％).Conclusions The public symphysis interval ≥ 23.36 mm or anterior interval of sacroiliac joint ≥ 9.82 mm can distinguish anteroposterior compression Ⅰ from Ⅱ injuries,and the combination of the two criteria can be beneficial to assessment of pelvic injury severity.

Spinocerebellar ataxias (SCAs), formerly known as autosomal dominant cerebellar ataxia, are a group of hereditary heterogeneous neurodegenerative disease that contains many subtypes. Spinocerebellar ataxia type 23 (SCA23), one type of SCAs, is caused by mutant prodynorphin (PDYN) gene. A 22-year-old patient was diagnosed with sporadic SCA23 due to gene detection, with a novel identified mutation, PDYN c.647C>T (p.P216L). Located in the dynorphin A-coding-region of PDYN gene, the pathogenic mechanism of the mutation may be relevant to the pathological changes caused by the variant including neurological dysfunction and death of cells. Mild improvement with the patient has been witnessed after active balance and speaking exercise.

OBJECTIVE: To explore the genetic mechanism underlying a case with para-Bombay phenotype. METHODS: The ABO and Lewis phenotype were identified with serological methods. The coding regions of exons 6 and 7 of the ABO and FUT1 genes were amplified with PCR and directly sequenced. Haploid sequence analysis was carried out on the variant sites of the FUT1 gene. RESULTS: Serological analysis confirmed that the proband has a rare para-Bombay phenotype. Direct sequencing revealed that he was a B.01/O.01.02 heterozygote for the ABO gene, and had heterozygous deletion for the 768 and 881-882 sites of the FUT1 gene. Further haploid analysis showed that the c.881_882delTT deletion has occurred in one haploid while c.768delC was present in the other haploid. The proband was therefore determined as a FUT1*01N.13/01N.20 heterozygote, which have resulted in frameshift in polypeptide chain p.Phe294Cysfs*40 and p.Val257Phefs*23, respectively. CONCLUSION A rare bi-allelic heterozygous deletion of para-Bombay phenotype has been identified in a blood donor. The c.881_882delTT and c.768delC deletions may decrease the activity of α-1,2-fucosyltransferase.
Animals ; Male ; ABO Blood-Group System/genetics* ; Alleles ; Fucosyltransferases/genetics* ; Genotype ; Heterozygote ; Mutation ; Phenotype ; Humans

Objective:To analyze the effect of meteorological conditions on mortality and population susceptibility of patients with acute myocardial infarction (AMI) in the Shantou area and to provide a scientific basis for the local public health system to prevent AMI.Methods:The AMI mortality data recorded in the resident cause of death surveillance database of Shantou from January 1, 2015, to December 31, 2020, were collected and the distribution lag nonlinear model was used to analyze the diurnal temperature range (DTR) and relative humidity (RH) on AMI mortality and the lag effect.Results:There were 13 932 deaths due to AMI in Shantou during the study period, with a male-to-female sex ratio of 1.3∶1. There was a significant association between high diurnal temperature difference exposure and low RH exposure and AMI deaths, with both single-day lag effects appearing and reaching a maximum at lag 2 day ( RR=1.019, 95% CI: 1.000-1.039; RR=1.018, 95% CI: 1.003-1.034); the cumulative lag effect was all maximal at lag 0-14 day ( RR=1.199, 95% CI: 1.025-1.401; RR=1.279, 95% CI: 1.117-1.465). The elderly (≥75 years) and female populations were susceptible to high DTR exposure and low RH exposure conditions. Conclusions:There was a significant association between DTR and RH and mortality of AMI in Shantou with a significant lag in their effects. Both female and elderly populations ≥75 years old were susceptible populations under high DTR and low RH exposure conditions.