Objective To investigate the thermal stability of solid potassium iodate and potassium iodate as additive in the sodium chloride,vitamin E,vitamin C and yellow prussiate.Methods HCR-2 type Differential Thermal Analyzer was used to carried out the differential thermal analysis of the potassium iodate and the potassium iodate in the sodium chloride,vitamin E,vitamin C and the yellow prussiate,and differential thermal curves were obtained and analyzed.Results The decomposition temperature of solid potassium iodate was 525℃ ; when mixed with sodium chloride,potassium iodate was stable below 300 ℃ ; vitamin C was unstable at 170-200 ℃ and underwent chemical changes; iodate and vitamin C underwent oxidation-reduction reaction at 145 to 160 ℃;potassium iodate with vitamin E at 300 ℃ was stable; yellow prussiate at 300 ℃ was stable; iodized salt was stable at cooking temperature below 300 ℃.Conclusions The potassium iodate has good stability below 525 ℃,however,potassium iodate iodized salt in the cooking process is easy to react with vitamin C in vegetables causing iodine losses,so iodized salt should be added just before the dish is done.

Objective To determine the immune responses induced by recombinant Salmonella ty-phimurium expressing the secreting antigen ESAT-6 of Mycobacterium tuberculosis. Methods ESAT-6 cod-ing gene was cloned and identified by PCR and sequencing. Prokaryotic expression plasmid pYA33-esat car-rying the ESAT-6 coding sequence was constructed firstly and electro-transformed into an attenuated strain X4550 of Salmonella typhimurium, the recombinant bacteria was named as X4550(33-esat). C57BL/6 mice were immunized intranasally (I. N) with 108 CFU recombinant bacteria at day 0 and 18. Cells from spleen, lung, mesenteric lymph node (MLN) and Peyer's patch (PP) were collected from mice after second immu-nization, and the specific IFN-γ-secreting cells and IL-4-secreting cells were detected by ELISPOT assay u-sing ESAT-6 peptide as stimulus. Furthermore, CTL effects were in vivo evaluated by CFSE assay. Results The results showed that cellular immune responses specific for ESAT-6 could be detected by ELISPOT assay. In lung and PP cells, immune responses against ESAT-6 were biased toward Th1 type, the frequency of IFN-γ-secreting cells was much higher than that of IL-4-secreting cells. In splenocytes and MLN cells, the anti-gen specific immune responses acted as Thl and Th2 balance, the frequency of IFN-γ-secreting cells was close to that of IL-4-secreting cells. CFSE assay indicated that recombinant bacteria could induce the high level of CTL effects specific for ESAT-6 peptide. Conclusion These results suggested that recombinant Sal-monella typhimurium X4550(33-esat) not only can induce cellular immune responses, but also can elicit specific CTL responses after I. N immunization. It also provided the useful information for the control of infec-tious disease of tuberculosis.

OBJECTIVETo investigate the feasibility of real-time elastography for quantitative evaluation of liver fibrosis in a rat model.

METHODSA total of 70 male Wistar rats were included in the group for dimethylnitrosamine (DMN)-induced liver injury, and 10 saline-injected rats were used as normal control. Hepatic injury was induced by a single intraperitoneal injection of DMN at a dose of 50 mg/kg of body weight. Nine or ten rats in the group with DNM injected and one or two rats in the normal control group were randomly selected and sacrificed at each of the following post-injection time: day 5, 7, 10, 14, 21, 24, and 28. And their livers were taken for pathology analysis. All the rats underwent real-time elastography before sacrificed in order to acquire area ratio of low-strain region (% AREA) and liver fibrosis index (LF index) which were compared with the stage of liver fibrosis and grade of necroinflammatory pathologically. By the different data, Spearman correlation analysis, rank-sum test or receiver operating characteristic curve was used.

RESULTSAmong 58 successfully modeled rats, there were nine, 13, 14 and 12 rats of S1, S2, S3 and S4 liver fibrosis on pathology, respectively, which were with or without mild necroinflammatory. The other 10 rats were found to be S0 with severe necroinflammatory. Values of LF index and % AREA both increased with liver fibrosis stage (P less than 0.05). There was certain correlation between LF index and liver fibrosis stage (r=0.643, P=0.000), so was % AREA and liver fibrosis stage (r=0.662, P=0.000). As for LF index, Areas under the receiver operating characteristic curve (Az) was 0.943, 0.890, 0.743 and 0.821 for the diagnosis of hepatic fibrosis S1 or higher, S2 or higher, S3 or higher and S4, respectively; as for % AREA, they were 0.948, 0.883, 0.772 and 0.842, respectively. However, we found a significant difference for LF index or % AREA between S0 with and without severe inflammatory activity rats (P=0.005 and P=0.017).

CONCLUSIONReal-time elastography is available for quantitative assessment of liver fibrosis in rats induced by DMN, but severe inflammatory activity can affect its accuracy.

Animals ; Dimethylnitrosamine ; adverse effects ; Elasticity Imaging Techniques ; Liver ; pathology ; Liver Cirrhosis, Experimental ; chemically induced ; pathology ; Male ; Rats ; Rats, Wistar

The herbs used as the material for traditional Chinese medicine are mostly planted at mountain areas where there is appropriate weather condition. Due to the complex terrain condition in mountain area, the planting at such places is always associated with a scattered pattern, which inevitably bring difficulty to the work of accurate measurement for the planting area. Advanced research about monitoring the planting area for medicine-herb under complex environmental condition is so critical and meaningful for developing the medicine-herb planting to a large scale and an industry. Therefore, this research, taking the planting of Herpetospermum pedunculosum in Luding county as an example, utilizes RS&GIS: using the image data from TM and ETM datum as the remote sensing information source, integrating GPS data from quadrat survey on the spots, and analyzing on a GIS platform, to extract the planting area of H. pedunculosum in Luding county. The results indicate that: (1) the method and technology used to estimate the planting area of H. pedunculosum are feasible and can be applied to monitoring the planting area of medicine-herb at mountain area under complex environmental conditions: (2) the planting area of H. pedunculosum in Luding county is estimated to be 857. 246 acres by the methods utilized in this research, which is closed to the value from local statistical data with an error range less than 5%; (3) in consideration of those inevitable errorrs generating from the procedures such as remote sensing data acquisition, data preprocessing, data interpretation, and so on, and the fact that the classification of remote sensing data, which must be implemented by those above procedures, was used directly to extract the herb type, the accuracy of area estimation for the herb is significantly influenced by the accuracy of classification for herb type, and the estimated area is inevitably associated with some error.
China ; Cucurbitaceae ; chemistry ; growth & development ; Geographic Information Systems ; Medicine, Tibetan Traditional ; Remote Sensing Technology ; methods

Primary health care institutions and physicians play a fundamental role and are the first line in the prevention and control of pandemics.Since the outbreak of coronavirus disease-2019(COVID-19) in 2019, nearly 4 million primary healthcare professionals across the country have been actively participated in pandemic screening, routine diagnosis and treatment, observation (or monitoring) isolated subjects, and crossing management.Facing outbreaks of pandemics, the key issue for primary health care institutions is how to improve the prevention and control capabilities, and how to take effective and comprehensive response measures.Based on the practical experiences of primary health care institutions in prevention and control for COVID-19 in China together with domestic and international experience and lessons in history, this paper discussed the following issues:internal and external structure of primary health care institutions, medical material storage and reservation, technical support, capability of medical personnel, and continuous health management in residents.This study aims to provide suggestions to improve the capability of primary health care institutions in pandemics prevention and control.

OBJECTIVETo report the results of clinical characteristics, enzyme activity determination and mutation analysis of GLB1 gene in a Chinese patient with mucopolysaccharidosis (MPS) type IVB (Morquio B disease).

METHODA 14-year-old Chinese boy with MPS type IVB was firstly diagnosed by blood leucocytes galactosamine-6-sulfate sulfatase (GALNS) and β-galactosidase (GLB1) determination, who was characterized by short stature, multiplex skeletal abnormalities, difficulty in walking. PCR-sequencing analysis was applied to detect the mutations in GLB1 of the patient.

RESULTThe patient was characterized by dwarfism, pectus carinatum, kyphosis, normal intelligence, and no neurologic damage of spasms, linguistic capacity and so on. The patient had normal GALNS enzyme activity and very low GLB1 enzyme activity [5.03 nmol/(h·mg) vs. normal value 118 - 413 nmol/(h·mg) ] in leukocytes. A compound heterozygous missense mutations c.442C > T(p.R148C)/c.1454A > G(p.Y485C) in GLB1 gene were detected in this patient. The mutation p.Y485C is a novel variant. With the method of gene analysis of new variant, the mutation p.Y485C was considered to be a pathogenic mutation.

CONCLUSIONThe MPS IVB patient showed severe multiple skeletal deformities, normal intelligence, no neurologic damage and very low GLB1 enzyme activity, who carries compound heterozygous mutations p.R148C/p.Y485C. The mutation p.Y485C in GLB1 gene may be a novel pathologic mutation of MPS type IVB.

Adolescent ; Amino Acid Sequence ; Asian Continental Ancestry Group ; genetics ; Chondroitinsulfatases ; genetics ; metabolism ; DNA Mutational Analysis ; Humans ; Joints ; pathology ; Male ; Molecular Sequence Data ; Mucopolysaccharidosis IV ; enzymology ; genetics ; pathology ; Mutation, Missense ; Pedigree ; Polymerase Chain Reaction ; Radiography ; Spine ; diagnostic imaging ; pathology ; beta-Galactosidase ; genetics ; metabolism

OBJECTIVETo investigate the high expression of HPV16L2N120E7E6 fusion protein by prokaryotic expression system, and evaluate its immunogenicity and antitumor efficacy in vaccinated mice.

METHODSThe HPV16L2N120E7E6 fusion gene, its codons were optimized to increase the expression of the protein, was constructed by overlap extension PCR and inserted into prokaryotic expression vector pET9a. Then the fusion protein was expressed by inducing with IPTG in E. coli strain BL21 (DE3) harboring with plasmid pETL2N120E7E6, and further detected by SDS-PAGE and Western-blot. Finally, the humoral and cellular immune responses were measured by ELISA and ELISPOT, respectively, in vaccinated mice with the purified HPV16L2N120E7E6 fusion protein, and the antitumor efficacy was assessed in mice using the TC-1 tumor challenge model.

RESULTSThe codon-optimized HPV16L2N120E7E6 fusion gene was highly expressed in E. coli strain BL21 (DE3) harboring with plasmid pETL2N120E7E6, and the amount of fusion protein was nearly 48.6% of the total bacterial protein. The purified fusion protein could induce high titer of specific antibody against L2, E7 and E6 in vaccinated mice. When accompanied with the adjuvant CpG, the fusion protein was able to elicit strong and moderate cellular immune responses in vaccinated mice against peptide HPV16E7(49-57) and peptide pools of HPV16E6, respectively. Furthermore, the tumor therapeutic experiment showed that HPV16L2N120E7E6 + CpG could prevent the tumor formation in 80.0% (8/10) vaccinated mice.

CONCLUSIONSThe data of this study suggest that HPV16L2N120E7E6 fusion protein could be a promising candidate vaccine for treatment of chronic HPV16 infection and post-operative adjuvant therapy for cervical cancer.

Adjuvants, Immunologic ; pharmacology ; Animals ; Cancer Vaccines ; immunology ; therapeutic use ; Capsid Proteins ; genetics ; immunology ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; Codon ; Escherichia coli ; immunology ; metabolism ; Female ; Humans ; Immunization ; methods ; Immunotherapy ; methods ; Mice ; Mice, Inbred C57BL ; Neoplasm Transplantation ; Oligodeoxyribonucleotides ; immunology ; Oncogene Proteins, Viral ; genetics ; immunology ; metabolism ; Papillomavirus E7 Proteins ; genetics ; immunology ; metabolism ; Papillomavirus Vaccines ; immunology ; therapeutic use ; Plasmids ; Recombinant Fusion Proteins ; genetics ; immunology ; metabolism ; Repressor Proteins ; genetics ; immunology ; metabolism

Objective To construct luciferase reporter plasmids of truncated fragments of different lengths of human guanylate binding protein 5(GBP5)gene promoter and analyze the transcriptional activity of each fragment to determine the core regulatory region.Methods GBP5promoter sequence was amplified by PCR,truncated into five fragments of different lengths and connected to pGL3-basic plasmid.The constructed recombinant plasmids pGL3-GBP5-11/21/31/41/51were transfected into 293FT cells and detected for luciferase activity.The binding sites of transcription factors in GBP5promoter region were predicted by JASPAR software,and Yin-Yang transcription factor 1(YY1)targeting the core regulatory region was selected and verified for the transcriptional regulatory activity.The CDS sequence of YY1 was amplified by PCR to construct the overexpression plasmid pIRES2-EGFP-YY1,which was then co-transfected to 293FT cells with plasmids pGL3-GBP5-21(-1 623 ～ +47 bp)and internal reference plasmid pRL-CMV,and detected for luciferase activity to analyze the regulation of transcription factor YY1 on GBP5 promoter activity.Results Colony PCR and double enzyme digestion identification proved that the plasmid of human GBP5 promoter reporter gene was correctly constructed;JASPAR software predicted that there were multiple transcription factor binding sites such as STAT1,YY1 and Foxp3 in GBP5promoter region.Double luciferase activity assay showed that pGL3-GBP5-21(-1 623 ～ +47 bp)showed the highest promoter activity,while the promoter activity of pGL3-GBP5-41(-520 ～ +47 bp)decreased significantly,suggesting that the core region of GBP5 promoter was located at upstream-1 623 ～-520 bp of 5 'UTR;Overexpression of YY1 significantly activated the GBP5 promoter activity and regulated the expression of GBP5.Conclusion The core regulatory region of human GBP5 promoter was located in upstream-1 623 ～-520 bp of the 5 'UTR,with a binding site of transcription factor YY1 existing in this region.Meanwhile,overexpression of YY1 significantly effected the activity of GBP5 promoter.

UNLABELLEDOBJECTIVE; To evaluate the clinical value of different CT diagnostic criteria for peripancreatic artery and vein invasion in pancreatic carcinoma through comparison with the findings on surgical exploration.

METHODSOf 72 patients of having suspected pancreatic carcinoma were examined by multiplane spiral CT. Among 43 confirmed by surgical pathology; 15 underwent pancreaticoduodenectomy; 28 were found to have unresectable tumors. The peri-pancreatic major vessels including the superior mesenteric artery, celiac artery, hepatic artery, superior mesenteric vein and portal vein were explored carefully during surgical exploration.

RESULTSThe criteria for peri-pancreatic artery invasion was the presence of one of the following signs: artery embeded in tumor, or more than half of the artery circumference involved by tumor with wall irregularity or stenosis. The sensitivity of the above described criteria was 75.0% (12/16). If the criteria of tumor involvement exceeding half of the vessel circumference were adhered to, the sensitivity was 87.5% (14/16), which was high than the former, but the specificity was lower than that of the former one (90.2% versus 95.1%). The criteria for peri-pancreatic vein invasion was presence of any of the following signs: vein obliteration, more than half of the vein circumference involved by tumor, vein wall irregularity, vein stenosis, tear-drop sign of superior mesenteric artery. The sensitivity of the above described criteria was 92.9% (39/42), higher than that of the criteria that more than half of the vessel circumference was involved by the tumor (69.0%, 29/42), but the specificity of both criteria was the same (97.4%, 37/38).

CONCLUSIONFor assessing peri-pancreatic artery and vein invasion, using the combination of different CT diagnostic criteria has higher accuracy than when using only criteria of more than half of vessel circumference involved by tumor.

Adult ; Aged ; Carcinoma, Pancreatic Ductal ; diagnosis ; surgery ; Celiac Artery ; diagnostic imaging ; Female ; Hepatic Artery ; diagnostic imaging ; Humans ; Male ; Mesenteric Artery, Superior ; diagnostic imaging ; Mesenteric Veins ; diagnostic imaging ; Middle Aged ; Neoplasm Invasiveness ; Pancreatic Neoplasms ; diagnosis ; surgery ; Pancreaticoduodenectomy ; Portal Vein ; diagnostic imaging ; Reproducibility of Results ; Sensitivity and Specificity ; Tomography, Spiral Computed ; methods

OBJECTIVEMany children were found to have low free carnitine level in blood by tandem mass spectrometry technology. In some of the cases the problems occurred secondary to malnutrition, organic acidemia and other fatty acid oxidation metabolic diseases, and some of cases had primary carnitine deficiency (PCD). In the present article, we discuss the diagnosis of PCD and evaluate the efficacy of carnitine in the treatment of PCD.

METHODWe measured the free carnitine (C0) and acylcarnitine levels in the blood of 270 000 neonates from newborns screening program and 12 000 children with suspected clinical inherited metabolic diseases by tandem mass spectrometry. The mutations of carnitine transporter protein were tested to the children with low C0 level and the diagnosis was made. The children with PCD were treated with 100 - 300 mg/kg of carnitine.

RESULTSeventeen children were diagnosed with PCD, 6 from newborn screening program and 11 from clinical patients. Mutations were found in all of them. The average C0 level [(2.9 ± 2.0) µmol/L] in patients was lower than the reference value (10 µmol/L), along with decreased level of different acylcarnitines. The clinical manifestations were diverse. For the 6 patients from newborn screening, 4 were asymptomatic, 1 showed hypoglycaemia and 1 showed movement intolerance from 2 years of age. For the 11 clinical patients, 8 showed hepatomegaly, 7 showed myasthenia, 6 showed cardiomyopathy, 1 showed chronic abdominal pain, and 1 showed restlessness and learning difficulty. Among these patients, 14 cases were treated with carnitine. Their clinical symptoms disappeared 1 to 3 months later. The C0 level in the blood rose to normal, with the average from (4.0 ± 2.7) µmol/L to (20.6 ± 8.3) µmol/L (P < 0.01). However, the level was still lower than the average level of healthy children [(27.1 ± 4.5) µmol/L, P < 0.01].

CONCLUSIONSeventeen patients were diagnosed with PCD by the test levels of free carnitine and acylcarnitines in blood with tandem mass spectrometry, and gene mutation test. Large dose of carnitine had a good effect in treatment of the PCD patients.

Cardiomyopathies ; diagnosis ; drug therapy ; genetics ; Carnitine ; analogs & derivatives ; blood ; deficiency ; genetics ; Child, Preschool ; DNA Mutational Analysis ; Female ; Follow-Up Studies ; Humans ; Hyperammonemia ; diagnosis ; drug therapy ; genetics ; Infant ; Infant, Newborn ; Male ; Muscular Diseases ; diagnosis ; drug therapy ; genetics ; Mutation ; Neonatal Screening ; methods ; Organic Cation Transport Proteins ; deficiency ; genetics ; Reference Values ; Tandem Mass Spectrometry