Objective:To evaluate the effect of nicastrin (nct) gene on the biological functions of melanocytes in zebrafish.Methods:By using a morpholino oligonucleotide (MO) technology, a nct-MO sequence targeting the zebrafish nct mRNA was designed, so was a MO control (ctrl-MO) sequence. Then, the enhanced green fluorescent protein (EGFP) mRNA with MO target sequence at its 5′ end was synthesized, and co-microinjected with the nct-MO or ctrl-MO sequence into the zebrafish embryos to verify the silencing efficiency of nct-MO and observe changes in developmental phenotypes in zebrafish. With wild-type zebrafish as a blank control group, real-time fluorescence-based quantitative PCR (RT-PCR) was conducted to determine the mRNA expression of melanin synthesis-and notch signaling pathway-related genes, including mitfa, tyr, tyrp1a, tyrp1b, dct, pmela, notch1a, notch1b and hey1 genes. One-way analysis of variance was used for the comparison of means among multiple groups, and least significant difference (LSD) - t test for multiple comparisons. Results:Eight hours after zebrafish fertilization, green fluorescence was observed in the zebrafish embryos in the ctrl-MO+EGFP mRNA group, but not in the nct-MO+EGFP mRNA group or blank control group. Forty-eight hours after fertilization, the proportion of pigmented area among the whole area of the tail of zebrafish larvae was significantly lower in the nct-MO group (0.169 ± 0.083) than in the ctrl-MO group (0.258 ± 0.042, t=3.202, P=0.005) , and disorderly pigment distribution in the tails was observed in the nct-MO group. RT-PCR revealed significant differences in the mRNA expression of pmela, tyrp1a and hey1 genes among the nct-MO group, ctrl-MO group and blank control group (all P < 0.05) , but no significant difference was observed in the mRNA expression of mitfa, tyr, tyrp1b, dct, notch1a or notch1b genes among the 3 groups (all P>0.05) ; the relative expression levels of pmela and tyrp1a mRNAs were significantly lower in the nct-MO group (0.708 ± 0.028, 0.558 ± 0.136, respectively) than in the ctrl-MO group (1.023 ± 0.142, 1.016 ± 0.134, respectively, both P < 0.05) . Conclusion:The nct gene may affect biological functions of melanocytes by regulating melanin synthesis in zebrafish.

OBJECTIVETo explore the effects of moxibustion with seed-sized moxa cone at "Ganshu" (BL 18) on liver furiction and morphology in rat with precancerous lesion of hepatic cellular cancer MCC).

METHODSA total of 60 male Wistar rats were randomly divided into a normal group (10 rats), a model group (20 rats), a 20-day treatment group (15 rats) and a 40-day treatment group (15 rats). HCC model was established by intraperitoneal injection of diethylnitrosamine (DEN). Rats in the normal group received no treatment. Rats in the model group were treated with fixation. Rats in the 20-day treatment group and 40-day treatment group were treated by moxibustion with seed-sized moxa cone at "Ganshu" (BL 18), once every other day, for 20 days and 40 days, respectively. Blood sample in each group was collected 1 d before model establishment, 20 d, 40 d and 84 d after model establishment. Chemical method was applied to test the activity of ALT (alamine aminotransferase), AST (aspartate transaminase) and GGT (glutamyl transpeptidase); at the end of model establishment, all the rats were sacrificed to observe the liver morphology changes.

RESULTSAfter the first therapeutic course, the. content of ALT and AST in the 20-day treatment group was significantly lower than that in the model group (all P<0. 05); after the second therapeutic course, the content of ALT, AST and GGT in the 40-day treatment group was insignificantly lower than that in the model group (all P>0. 05). Under light microscope, the slice of liver tissue indicated that primary tumor was induced in the model group, and the tumor cells were stained and irregular; the cytoplasm in the 20-day treatment group was even, and the tumor cells were few with several nodules alone. In the 40-day treatment group the liver morphology was normal and the staining was even; the tumor cells were few without nodules or a few. Conclusion Moxibustion with seed-sized moxa cone at "Ganshu" (BL 18) could reduce the serum content of ALT, AST and GGT in rats with HCC, which could protect the liver and: delay the DEN-induced precancerous lesion on some levels.

Acupuncture Points ; Animals ; Aspartate Aminotransferases ; blood ; Humans ; Liver ; pathology ; Liver Neoplasms ; enzymology ; pathology ; therapy ; Male ; Moxibustion ; Rats ; Rats, Wistar ; gamma-Glutamyltransferase ; blood

Objective:To establish a presenilin enhancer-2 (PSENEN) gene-silenced human immortalized keratinocyte (HaCaT) cell model, and to evaluate the effect of PSENEN gene silencing on the proliferation of and γ-secretase expression in HaCaT cells.Methods:Three shRNAs targeting the PSENEN gene were constructed, and inserted into the linearized LV3-pGLV-h1-GFP-puro vector to establish a recombinant lentiviral expression plasmid. After restriction enzyme digestion and sequencing, lentiviral packaging and purification were performed, and lentiviral titer was determined. Cultured HaCaT cells were divided into 5 groups: shRNA1, shRNA2 and shRNA3 groups treated with the lentivirus solutions containing PSENEN gene-targeted shRNA1, shRNA2 and shRNA3 respectively, NC group treated with the lentivirus solution containing a negative control shRNA (shNC) , and blank group treated without lentivirus solution. After transfection, inverted fluorescence microscopy was performed, and transfection efficiency was determined by flow cytometry. Cell counting kit-8 (CCK8) assay was performed to evaluate the effect of PSENEN gene silencing on the proliferation of HaCaT cells, and real-time fluorescence-based quantitative PCR (qPCR) and Western blot analysis were conducted to determine the mRNA and protein expression of PSENEN, nicastrin (NCT) , presenilin-1 (PS1) and anterior pharynx defective 1a (APH1a) genes respectively. Statistical analysis was carried out by using repeated measures analysis of variance, one-way analysis of variance, and least significant difference t test for multiple comparisons. Results:Inverted fluorescence microscopy showed that fluorescence was observed in the shRNA1 group, shRNA2 group, shRNA3 group and NC group, and flow cytometry showed that the transfection efficiency was over 98% in the above 4 groups. qPCR and Western blot analysis revealed that the mRNA and protein expression of PSENEN gene significantly decreased in the shRNA1 (0.187 ± 0.010, 0.219 ± 0.097, respectively) , shRNA2 (0.163 ± 0.022, 0.208 ± 0.014, respectively) and shRNA3 (0.174 ± 0.009, 0.185 ± 0.062, respectively) groups compared with the NC group (1.054 ± 0.272, 1.076 ± 0.075, respectively, all P < 0.001) . CCK8 assay showed that the cellular proliferative activity significantly increased in the shRNA1 group compared with the NC group at 0, 12, 36 and 48 hours (all P < 0.05) , and there was no significant difference between the 2 groups at 24 or 60 hours (both P > 0.05) ; the cellular proliferative activity was significantly higher in the shRNA2 and shRNA3 groups than in the NC group at 0, 12, 24, 36, 48 and 60 hours (all P < 0.05) . There was no significant difference in the mRNA expression of NCT, PS1 and APH1a genes among the shRNA1 group, shRNA2 group, shRNA3 group, NC group, and blank group ( F= 8.168, 4.644, 1.981, respectively, all P > 0.05) , while the relative protein expression level of mature NCT (mNCT) , immature NCT (imNCT) , carboxyl-terminal fragment of PS1 (PS1-CTF) and APH1a significantly differed among the above 5 groups ( F= 39.268, 5.929, 27.842, 20.663, respectively, all P ≤ 0.01) . Compared with the NC group, the shRNA1, shRNA2 and shRNA3 groups all showed significantly decreased protein expression of mNCT, PS1-CTF and APH1a (all P < 0.01) , but insignificant changes in imNCT protein expression (all P > 0.05) . Conclusion:The PSENEN gene-silenced HaCaT cell model was successfully constructed, and the PSENEN gene silencing could lead to an increase in the cellular proliferative activity of HaCaT cells and a decrease in the protein expression of γ-secretase subunits mNCT, PS1-CTF and APH1a.

Binding Sites ; Histone-Lysine N-Methyltransferase ; chemistry ; metabolism ; Histones ; chemistry ; metabolism ; Humans ; Methylation ; Multiprotein Complexes ; chemistry ; metabolism ; Nuclear Proteins ; chemistry ; metabolism ; Protein Binding ; Protein Conformation ; Yeasts ; chemistry

Objective: To analyze the delay on detection, care seeking, diagnosis and treatment of tuberculosis among students in Inner Mongolia Autonomous Region (Inner Mongolia) from 2011 to 2022 and its influencing factors, so as to provide support for the prevention and treatment of tuberculosis among students. Methods: The general demographic indicators of students with tuberculosis in Inner Mongolia from January 1, 2011 to December 31, 2022 were collected from the infectious disease monitoring (new) module of the China Disease Prevention and Control Information System. General characteristics and trend of four types of delayed pulmonary tuberculosis patients in students were analyzed. The influencing factors were analyzed using univariate and multivariate Logistic regressions. Results: From 2011 to 2022, there were 6 032 cases of pulmonary tuberculosis among students in Inner Mongolia. The rates of delayed detection, delayed care seeking, delayed diagnosis, and delayed treatment were 51.71%, 64.01%, 7.82 %, and 2.30%, respectively. The results of multivariate Logistic regression analysis showed that tracking ( OR =1.51) in the patient source,league level diagnosis ( OR =3.16) in the diagnostic institution level,and county level diagnosis ( OR =2.41) were positively associated with delayed discovery ( P <0.05). At the level of diagnostic unit, league city level diagnosis ( OR =2.69), and county level diagnosis ( OR =3.67) associated with more delayed care seeking ( P <0.05). Referral ( OR =1.58) and follow up ( OR =2.55), floating population ( OR =2.05), further consultation with a doctor ( OR =2.11), and no results in imaging manifestations ( OR =2.19) were positively associated with delayed diagnosis( P <0.05). The factors contributing to delayed treatment were referral ( OR =1.84), follow up ( OR =4.91), active screening ( OR =5.46), and floating population( OR =1.95)( P <0.05). Conclusions From 2011 to 2022, the delay on detection and care seeking for tuberculosis patients among students in Inner Mongolia is at a relatively high level, while the delay in diagnosis and treatment is at a relatively low level but shows an increasing trend. It is necessary to focus on the factors associated with delays in identification, diagnosis and treatment in tuberculosis outbreak in the context of school to prevent or reduce school tuberculosis outbreak.

【Objective】 To understand the occupational exposure of blood center staff during blood collection and supply, analyze the influencing factors, explore preventive measures and improve the self-protection awareness of staff, so as to further improve the prevention of such occupational exposure in blood centers. 【Methods】 Seventy-two occupational exposure cases, which occurred and registered in Beijing Red Cross Blood Center from January 2010 to December 2021, were reviewed by retrospective survery methods, and such cases were classified and analyzed statistically. The incidence of blood-borne occupational exposure was compared by genders, departments, occurrence locations, education level, occupation type, work link, causes of exposure and body parts, so as to analyze the influencing factors, and then propose corresponding prevention strategies. 【Results】 From 2010 to 2021, a total of 72 blood-borne occupational exposure cases occurred. When classified by gender, the exposure rate of women was more than twice that of men; by age, the exposure rate of 18~25 years old was the highest, reaching 7.84%, but 26~40 years old group contributed the most cases, accounting for 70.83% (51/72); by working years, the exposure rate of staff with working years less than 3 years was relatively high, but the staff with 4~10 working years contributed the most cases, accounting for 54.17% (39/72); by the education level, the the exposure rate of stall with bachelor degree and below was the highest(90.28%, 65/72); by the professional title level, the exposure rate of staff with primary and intermediate professional titles was relatively high (94.44% , 68/72); by departments, the exposure rate of Blood Donation Service was the highest (81.94%, 59/72), which was twice or even three times than that of other departments; by occupational type, nurses had the highest exposure rate, accounting for 87.5% (63/72); by body parts, hands were the most vulnerable body parts to occupational exposure; by location, the exposure rate on mobile vehicles is four times than that in indoor places; by the number of exposure times, the incidence of one exposure cases accounted for 72.22%(52/72)while multiple exposure was rare by work link, the exposure rate of the sampling link was significantly higher than that of the other links. 【Conclusion】 Blood-borne occupational exposure of staff in blood centers is closely related to gender, age, working years, professional title level, department, occupational type, occurrence place, and body part, and has nothing to do with the education level. More attention should be paid to middle-aged female nurses on the front line of blood donation service, staff with primary or intermediate titles and undergraduate degrees. Otherwise, training and protection should be enhanced, operation methods be standardized, the blood collection environment be improved, and the management, automation and intelligent level of blood collection be promoted.

The fatty alk(a/e)ne biosynthesis pathway found in cyanobacteria gained tremendous attention in recent years as a promising alternative approach for biofuel production. Cyanobacterial aldehyde-deformylating oxygenase (cADO), which catalyzes the conversion of Cn fatty aldehyde to its corresponding Cn-1 alk(a/e)ne, is a key enzyme in that pathway. Due to its low activity, alk(a/e)ne production by cADO is an inefficient process. Previous biochemical and structural investigations of cADO have provided some information on its catalytic reaction. However, the details of its catalytic processes remain unclear. Here we report five crystal structures of cADO from the Synechococcus elongates strain PCC7942 in both its iron-free and iron-bound forms, representing different states during its catalytic process. Structural comparisons and functional enzyme assays indicate that Glu144, one of the iron-coordinating residues, plays a vital role in the catalytic reaction of cADO. Moreover, the helix where Glu144 resides exhibits two distinct conformations that correlates with the different binding states of the di-iron center in cADO structures. Therefore, our results provide a structural explanation for the highly labile feature of cADO di-iron center, which we proposed to be related to its low enzymatic activity. On the basis of our structural and biochemical data, a possible catalytic process of cADO was proposed, which could aid the design of cADO with improved activity.
Aldehyde Oxidoreductases ; chemistry ; genetics ; metabolism ; Amino Acid Sequence ; Amino Acid Substitution ; Bacterial Proteins ; chemistry ; genetics ; metabolism ; Binding Sites ; Biocatalysis ; Crystallography, X-Ray ; Gas Chromatography-Mass Spectrometry ; Ligands ; Molecular Dynamics Simulation ; Molecular Sequence Data ; Protein Structure, Tertiary ; Sequence Alignment ; Synechococcus ; enzymology

To explore the application of gene chip technology in the acupuncture research so as to provide evidences for the mechanism of acupuncture for regulating bodies. The literature on the application of gene chip technology in the acupuncture field from 2001 to 2016 was collected in PubMed, Springer, CNKI and WANFANG databases, which was analyzed and summarized. There were some achievements of the technology for acupuncture research, focusing on the five aspects, including the study of the relationship between meridian-point and viscera, the influencing factors of acupuncture effect, the effect and mechanism of acupuncture analgesia, the mechanism of acupuncture anti-aging, the effect and mechanism of acupuncture for diseases of each system. Gene chip technology plays an important role in researching acupuncture mechanism. It is an important technology for genomics study of acupuncture. However, there are also some disadvantages such as high cost, deficient data mining, non-uniform observation objects, deficient professionals, etc. All those need further resolution so as to promote the application of this technology in the acupuncture researching field.

There are various contents about diagnosis and treatment of low back pain in(), which are closely related to meridians and collaterals, forming a complete theoretical system of meridian differentiation for low back pain. Its theory and application are interpreted in this paper from five main aspects, including 18 meridians and collaterals densely distributing in the tendons and bones of the waist, the pathogenesis of low back pain lying in the disorders of the meridianand blood, the nature of low back pain being reflected by the color and shape of meridians and collaterals, the disordered meridians of low back pain being first identified based on syndrome differentiation, the acupuncture prescription being selected according to meridian differentiation. The purpose of this paper is to guide clinical practice, explore the advantages and characteristics of meridian differentiation, and provide theoretical reference and evidence for promoting the standardization of TCM.