To obtain and analyze the sequence of the nucleocapsid gene from bovine coronavirus, and to produce the fusion protein of the N gene in E.coli in order to use this recombinant protein for the study of bovine coronavirus. The N gene of BCV-DQ strain was amplified by RT-PCR, in which the primers were designed on the basis of N gene sequence of BCV-Mebus strain. The PCR products of 1 347 bp in length were cloned and sequenced, and then inserted into the prokaryotic vector pET30a. The recombinant plasmids were then transformed into Escherichia coli BL21 and identified by SDS-PAGE and Western blot assay. ELISA assay was optimized of N protein as the coating antigen to detect the viruses in the clinical samples. In comparison with 6 BCV strains in GenBank, the sequence identity was proved to be more than 98.3%. Result in SDS-PAGE showed that the fusion protein had a molecular weight of 60 ku, and could be specifically recognized by mouse serum against BCV. The indirect ELISA was used to test 256 serum samples collected from Heilongjiang province and 65.23% samples were positive. On testing field samples, an overall agreement of 95.31% was generated between the the neutralization test of viruses (VN) and indirect ELISA. It is apparent that the N gene was highly conservative and is expressed in E. coli in high level,also the prokaryotic expression products of this gene show a fine reactiongenicity in immune responses. It was also suggested that the N protein may be a useful antigen for sero-diagnosis and epidemiological investigation of BCV.

Objective:To evaluate the effectiveness of Minqing Ao dental desensitizer in the treatment of early childhood caries (ECC).Methods:41 0 teeth with ECC in 1 1 0 cases were divided into 3 groups.Minqing Ao dental desensitizer,fluoride toothpaste and ordinary toothpaste without fluoride were respectively used in group A,B and C.The laser fluorescence value(LFV)was meas-ured and compared before treatment,2,4,6 weeks and 6 months after treatment.Results:2 weeks after treatment,the LFV was not significantly changed in the 3 groups.4 weeks after treatment LFV in group A was decreased(P<0.01 ),the LFV in group A was lower than that in group B(P<0.05)and in group C(P<0.01 ).While there was no significant difference between group B and C(P>0.05).6 weeks after treatment LFV in group A and B was decreased(P<0.01 and P<0.05),there was significant difference be-tween each 2 groups(P<0.05).6 weeks and 6 months after treatment LFV in group A was lower than that in group B(P<0.01 ),and in group B was lower than that in group C(P<0.01 ).Conclusion:Minqing Ao is effective for treatment of early childhood caries.

Objective To study the natural developing characteristics of language skill of the autistic children. Methods 300 untreated autism children were included. Their developmental characteristics of individuals were evaluated by the Psycho-Educational Profile-Revised (PEP-R). Results The developmental quotient of most children (89.33%) was less than 70. And the developmental quotient gradually decreased with age (P<0.01). The scores of language performance and language verbal were deviation (P<0.01). Conclusion Language characteristics of autistic children without treatment is abnormal in development progress and order, and the situation would be worse with age.

Objective To analyze the clinical outcome of concurrent ehemo-radiotherapy in stage Ⅳ non-small cell lung cancer(NSCLC).Methods From Jan.1997 to Dec.2006,214 patients with patho logically or cytologically proven stage Ⅳ NSCLC were included in this analysis.Of those patients,98 re ceived radiotherapy concurrently with 3-week cycle chemotherapy(group A),18 received radiotherapy con currently with weekly chemotherapy(group B) ,44 received chemotherapy alone,37 received radiotherapy a lone and 13 received sequential chemo-radiotherapy.The primary tumor was treated by three-dimensional conformal radiotherapy(3DCRT) or conventional radiotherapy with conventional fraefionation or late-course accelerated hyperfraction (LA H RT).Group A received 21-28 days cycle cisplatin-based chemotherapy (cis platin combined with PTX,DTY,NVB or Vp-16) ,and group B received weekly DDP combined with PTX or topteeon for 4-6 weeks.Results The follow-up rate was 99%.The 1-and 2-year overall survival rates of group A,group B,chemotherapy alone,radiotherapy alone and sequential chemo-radiotherapy were 41% and 11% ,16% and 0,31% and 7% ,34% and 10% ,26% and 3% ,respectively(x2 = 11.18,P=0.025).The patients with concurrent 3DCRT,LAHRT and radiotherapy dose≥70 Gy had better survival in group A than those in chemotherapy alone group.Patients who received≥2 cycles chemotherapy with concurrent radio therapy had longer survival time than those who had ≥2 cycles chemotherapy alone. Conclusions Con current chemotherapy and 3DCRT,LAHRT with the dose ≥70 Gy can improve the overall survival of patients with stage Ⅳ non-small cell lung cancer.

Objective:To explore the effect of adipose derived mesenchymal stem cells to regulate the differentiation of macrophage RAW264.7.Methods:First,we used RAW264.7 cells to simulate macrophage and induced them to M 1 macrophage with lipopolysaccharide ( LPS,1 μg/ml) .Then we cultured these RAW264.7 cells in culture mediums which were previously used to culture adipose derived mesenchymal stem cells to imitate the transplantation of ADMSC .Last,the mRNA relative expression of IL-10, IGF-1,Arg-1,TNF-α,FIZZ1,SPHK-1 was detected by real-time PCR.The protein expression of IL-12 p40,IL-27 Rα,IL-10 was detected by Western blot.Results:After been cultured in ADMSCCM and induced by LPS ,M1 markers (TNF-αmRNA,IL-12 p40;P<0.05) of the RAW264.7 cells declined while M2 markers (IGF-1 mRNA,IL-10 mRNA,IL-10;P<0.05) rose.Conclusion: ADMSC can secrete soluble cytokines to induce the RAW264.7 cell,which have been induced to the M1 macrophages,to differentiate towards M2 macrophages.

Objective:To establish the reference value of the angle between the left ventricular inflow and outflow tract (LIOA) in normal fetuses in the second and third trimesters, and observe the correlation between fetal LIOA and gestational age, cardiac axis, cardiac size, and estimated fetal weigh (EFW).Methods:Fetal LIOA in normal fetuses with gestational age from 16 weeks to 39 + 6 weeks were obtained prospectively by two-dimensional ultrasound in the Second Xiangya Hospital from November 2022 to April 2023. Pearson′s correlation coefficient was used to analyze the correlation between fetal LIOA and gestational age, cardiac axis, cardiovascural size and EFW. Results:The LIOA range of 651 normal fetuses was (44.39±7.67)°, and it was found that LIOA was not related to gestational age. LIOA mildly positively correlated with the cardiac axis ( r=0.22, P<0.05) while not correlated with gestational age, cardiovascural size or EFW (all P>0.05). Conclusions:The range of LIOA in normal fetuses were established. Fetal LIOA is constant in the second and third trimesters and it is mildly positively correlated with the cardiac axis. Evaluating fetal LIOA may also provide information for future research on the fetal aortic hemodynamic development.

Objective To report a pedigree with X?linked dominant protoporphyria(XLDPP), and to detect 5?aminolevulinic acid synthetase 2(ALAS2)gene mutations in this pedigree. Methods A clinical investigation was performed in a pedigree with XLDPP, and relevant data were collected from family members. A next?generation sequencing method was applied to screen possible mutation sites, and Sanger sequencing was performed to determine pathogenic gene mutations. Dermoscopy was conducted to observe skin lesions in the patients with XLDPP, and the Fotofinder system and very high frequency (VHF) ultrasound system were utilized to assess the severity of photodamage. Liver and gallbladder ultrasonography as well as blood examination were performed for all the family members. Results A deletion mutation, c.1706?1709ΔAGTG, was detected in the ALAS2 gene on the X chromosomes of all the patients in this family, which led to replacement or loss of 19-20 C?terminal residues through transcriptional frameshifting, and eventually caused an increase in ALAS2 activity. In the patients with XLDPP, skin photodamage was relatively severe;protoporphyrin?induced hepatobiliary damage was observed and aggravated with age;anemia and iron deficiency occurred sometimes. Conclusion The deletion mutation c.1706?1709ΔAGTG of the ALAS2 gene may be the underlying cause of XLDPP in this pedigree.

OBJECTIVE: Thinking on the construction of the medical device type archives information system. METHODS: This paper introduces the concept and significance of medical device variety archives, and puts forward the overall construction idea and system framework of medical device variety archives by analyzing its construction difficulties. RESULTS: Considering the long-term nature and complexity of the construction of medical device variety archives, the system can be constructed in accordance with the three steps of system building, platform building and data management, and the overall technical architecture can be designed from the eight aspects of user layer, business application layer, application support layer, data resource layer, infrastructure layer, security, standards and operation and maintenance management. CONCLUSIONS Architecture design is the foundation of system construction, and its design rationality is very important for the success of system construction. The architecture design proposed in this study has a certain reference role for promoting the construction of medical device variety archives management system.
Information Systems ; Reference Standards

Chang-Kang-Fang (CKF) formula, a Traditional Chinese Medicine (TCM) prescription, has been widely used for the treatment of irritable bowel syndrome (IBS). However, its potential material basis and underlying mechanism remain elusive. Therefore, this study employed an integrated approach that combined ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q/TOF-MS) with network pharmacology to systematically characterize the phytochemical components and metabolites of CKF, as well as elucidating its underlying mechanism. Through this comprehensive analysis, a total of 150 components were identified or tentatively characterized within the CKF formula. Notably, six N-acetyldopamine oligomers from CicadaePeriostracum and eight resin glycosides from Cuscutae Semen were characterized in this formula for the first time. Meanwhile, 149 xenobiotics (58 prototypes and 91 metabolites) were detected in plasma, urine, feces, brain, and intestinal contents, and the in vivo metabolic pathways of resin glycosides were elaborated for the first time. Furthermore, network pharmacology and molecular docking analyses revealed that alkaloids, flavonoids, chromones, monoterpenes, N-acetyldopamine dimers, p-hydroxycinnamic acid, and Cus-3/isomer might be responsible for the beneficial effects of CKF in treating IBS, and CASP8, MARK14, PIK3C, PIK3R1, TLR4, and TNF may be its potential targets. These discoveries offer a comprehensive understanding of the potential material basis and clarify the underlying mechanism of the CKF formula in treating IBS, facilitating the broader application of CKF in the field of medicine.
Humans ; Tandem Mass Spectrometry/methods* ; Irritable Bowel Syndrome/drug therapy* ; Molecular Docking Simulation ; Drugs, Chinese Herbal/chemistry* ; Glycosides ; Chromatography, High Pressure Liquid/methods*