Clinical application differences between TW3-Carpal and TW3-RUS based artificial intelligence-assisted bone age assessment system: a real-world pilot study / 中华放射学杂志

Objective:To investigate the differences between Tanner-Whitehouse (TW)3-Carpal and TW3-RUS(radius, ulna and short bone)-based artificial intelligence (AI)-assisted bone age assessment system using real world data.Methods:The image data of 262 children who received X-ray examination of left wrist in the Affiliated Children′s Hospital, Capital Institute of Pediatrics from July to September 2021 were retrospectively collected. The AI bone age assistant methods based on TW3-RUS and TW3-Carpal criteria were used to obtain the bone age results, respectively. Two senior pediatric radiologists evaluated the bone age on the basis of TW3-RUS and TW3-Carpal criteria, and the averaged values of two reviewers was calculated and taken as the gold standard reference. The cases were stratified into six age groups at 3-year intervals according to the gold standard reference, including 1-3 ( n=10), 4-6 ( n=35), 7-9 ( n=70), 10-12 ( n=118), 13-15 ( n=27) and 16-18 ( n=2) years old groups. Intraclass correlation coefficient (ICC) was used to evaluate the consistency between AI results and the gold standard bone age results. Pearson correlation method was used to measure the reliability between AI results and the gold standard results. The difference of bone age results between using TW3-RUS and TW3-Carpal criteria in different age groups was compared using paired t-test. Results:As for the whole sample, the results based on TW3-RUS criteria were 8.9±3.1 years old for AI assessment and 8.7±2.9 years old for the golden standard reference, with the ICC of 0.983; and the results based on TW3-Carpal criteria were 8.7±3.0 years old for AI and 8.8±2.8 years old for the golden standard reference, with the ICC of 0.976. Positive correlation were found in both TW3-RUS ( r=0.985, P<0.001) and TW3-Carpal criteria groups ( r=0.978, P<0.001). There were significant differences between TW3-RUS and TW3-Carpal at age groups of 7-9( t=-3.36, P=0.001), 10-12( t=-1.77, P=0.046), and 13-15 years old ( t=1.84, P=0.040). The bone age assessment using TW3-RUS and TW3-Carpal criteria were both in good agreement with the gold standard reference in age group of 4-6 years old (ICC=0.929 and 0.940), as well as in age group of 7-9 years old (ICC=0.882 and 0.927, respectively), with the results using TW3-Carpal criteria were slightly higher. As for the age groups of 10-12 and 13-15 years old, the method using TW3-RUS criteria showed excellent agreement with the gold standard reference (ICC=0.962 and 0.963, respectively), which were better than the performance of method using TW3-Carpal criteria (ICC=0.744 and 0.605, respectively). Conclusions:AI-assisted bone age system based TW3-Carpal and TW3-RUS criteria both show good reliability and accuracy in the bone age measurements. The AI method based TW3-Carpal criteria shows better performance in age group of 4-9 years old, while the method based on TW3-RUS criteria may be better for children of age 10-15 years old.

Research on rapid detection technology of airborne pathogenic organisms based on up-conversion phosphor technology / 中华预防医学杂志

Objective: To explore the application of up-conversing phosphor technology (UPT) to detect pathogenic organisms in the air. Methods: The performance of UPT was verified with Staphylococcus aureus, Yersinia pestis and Escherichia coli O157 as simulated strains, including stability, specificity, sensitivity and response time tests; Air particle sampler is used to collect air samples in the field microenvironment test chamber, and UPT is used for detection. At the same time, compared with the traditional culture method, the practicability of UPT is verified. Results: The coefficient of variation in laboratory was 9.62% and 8.02% when the concentration of 107 CFU/ml and 108 CFU/ml were detected by UPT. The results were less than the allowable target, and the detection system had good stability. The specificity of UPT was verified by Staphylococcus aureus. The results showed that no non-Staphylococcus aureus was detected, and the positive detection rate of different kinds of Staphylococcus aureus was 100%. The specificity of the detection system was good. The sensitivity of UPT for detecting Staphylococcus aureus was 104 CFU/ml. Detection sensitivity of Yersinia pestis ≥103 CFU/ml; The detection sensitivity of Escherichia coli O157 is ≥103 CFU/ml, and the response time of UPT to bacteria is within 15 min (all 10 min 15 s). The detection results of bacteria contentration in the air of the on-site microenvironment test cabin by UPT showed that when the concentration of Escherichia coli O157 in the air reached above 104 CFU/m3, the detection results of UPT were positive, and with the increase of air concentration, the numerical concentration measured by UPT showed an upward trend, which was positively correlated with the concentration of bacteria in the air. Conclusion: UPT may be feasible as a rapid method to evaluate the species and contentration of pathogenic organisms in the air.

Hydnocarpin inhibits malignant progression of triple negative breast cancer via CNOT4-mediated ubiquitination and degradation of YAP / 中国中药杂志

This study aims to investigate the effect and mechanism of hydnocarpin(HC) in treating triple negative breast cancer(TNBC). Cell counting kit-8(CCK-8), xCELLigence real-time cellular analysis(RTCA), and colony formation assay were employed to determine the effects of HC on the proliferation of two TNBC cell lines: MDA-MB-231 and MDA-MB-436. The effects of HC on the migration and invasion of TNBC cells were detected by high-content analysis, wound-healing assay, and Transwell assay. The changes in the epithelial-mesenchymal transition(EMT) and the expression of invasion-and migration-associated proteins [E-cadherin, vimentin, Snail, matrix metalloproteinase-2(MMP-2), and MMP-9] were detected by Western blot. Western blot and RT-qPCR were employed to determine the protein and mRNA levels of Yes-associated protein(YAP) and downstream targets(CTGF and Cyr61). TNBC cells were transfected with Flag-YAP for the overexpression of YAP, and the role of YAP as a key target for HC to inhibit TNBC malignant progression was examined by CCK-8 assay, Transwell assay, and wound-healing assay. The pathway of HC-induced YAP degradation was detected by the co-treatment of proteasome inhibitor with HC and ubiquitination assay. The binding of HC to YAP and the E3 ubiquitin ligase Ccr4-not transcription complex subunit 4(CNOT4) was detected by microscale thermophoresis(MST) assay and drug affinity responsive target stability(DARTS) assay. The results showed that HC significantly inhibited the proliferation, colony formation, invasion, and EMT of TNBC cells. HC down-regulated the protein and mRNA levels of CTGF and Cyr61. HC down-regulated the total protein level of YAP, while it had no effect on the mRNA level of YAP. The overexpression of YAP antagonized the inhibitory effects of HC on the proliferation, migration, and invasion of TNBC cells. HC promoted the degradation of YAP through the proteasome pathway and up-regulated the ubiquitination level of YAP. The results of MST and DARTS demonstrated direct binding between HC, YAP, and CNOT4. The above results indicated that HC inhibited the malignant progression of TNBC via CNOT4-mediated degradation and ubiquitination of YAP.

Study on Periplaneta americana Polypeptide PAE2 in Reversing Multidrug Resistance for Liver Cancer / 中国实验方剂学杂志

Objective:To investigate the effect and mechanism of PAE₂， a polypeptide of <italic>Periplaneta americana， </italic>in reversing multidrug resistance （MDR） for liver cancer <italic>in vivo</italic>. Method:Balb/c-nude mice were inoculated with HepG2 and HepG2/ADM cells under the armpits to establish animal models of liver cancer sensitive strains and animal models of MDR respectively. After successful modeling， the nude mice were randomly divided into normal group， HepG2 model group， HepG2/ADM model group， sorafenib group （positive drug control group， <italic>ig</italic> 30 mg·kg^-1）， HepG2/ADM+PAE₂ （<italic>iv</italic>） low， medium and high dose groups （50， 100， 200 mg·kg^-1）， HepG2/ADM+PAE₂ （<italic>ig</italic>） low， medium， and high dose groups （50， 100， 200 mg·kg^-1）， skim cream group （<italic>ig</italic> 200 mg·kg^-1）， and CⅡ-3 group （<italic>ig</italic> 200 mg·kg^-1）， all of which received corresponding drug treatment. The body weight and tumor volume of nude mice were measured and recorded every 2 days. The next day after the last administration， tumor tissues of nude mice were taken to record the tumor weight. The effect of <italic>P. americana </italic>polypeptide PAE₂ on permeability-glycoprotein（P-gp）， lung resistance protein（LRP） ， breast cancer resistance protein（BCRP）， protein kinase C（PKC）， glutathione S-transferase-π（GST-π）， topo-isomerase typeⅡ（ToPoⅡ）， multidurg resistance gene 1（MDR1）and Multidrug resistance-associated proteins（MRP1） of the protein level and gene level expression in tumor tissues were determined by immunohistochemistry （IHC） and real-time quantitative polymerase chain reaction （Real-time PCR）. In addition， both oral and intravenous administration groups were set up at the same time for preliminary study on the basic pharmacokinetic characteristics of <italic>P. americana </italic>polypeptide PAE₂. Result:After the successful modeling， the body weight of the nude mice was significantly lower than that in the normal mice（<italic>P</italic><0.05）. After treatment with corresponding drugs， the body weight increased to a certain extent， but it was still not as good as the normal nude mice. In <italic>iv</italic> administration， the medium-dose <italic>P. americana </italic>polypeptide PAE₂ showed the best anti-tumor effect as compared with the model group （<italic>P</italic><0.05）， while in oral administration， the anti-effect increased with the increase of the dose， so the high-dose group showed the best effect （<italic>P</italic><0.05）. Preliminary crude extract CII-3 had no obvious anti-tumor effect， and skim cream showed a certain anti-tumor effect （<italic>P</italic><0.05）. <italic>P. americana </italic>polypeptide PAE₂ had certain effects on MDR related proteins and enzymes<italic> in vivo</italic>， mainly by inhibiting the expression of LRP and BCRP in tumor tissues and affecting the expression of these related proteins and genes to different degrees to inhibit intracellular drugs outflow， thereby promoting tumor apoptosis， and the effect was superior to that of the <italic>P. americana</italic> crude extract CⅡ-3 and skim cream. Conclusion:<italic>P. americana</italic> polypeptide PAE₂ may reduce the drug efflux， promote intracellular drug accumulation and apoptosis by affecting the expression of related proteins and enzymes that mediate multidrug resistance， thereby exerting a reverse effect on HepG2/ADM cells Balb/c MDR in nude mice.

Calculation of Likelihood Ratio for Identifying Half Sibling Relationship When Both Biological Mothers Participate / 法医学杂志

Objective To derive the formulae for likelihood ratio （LR） calculation of half sibling relationships when both mothers participate. Methods Based on the fact that both biological mothers participate in the identification of half sibling relationship between the two individuals, test hypothesis for the identification of half sibling relationship was established. Conditional probability ratios of genetic evidence under null hypothesis and alternative hypothesis conditions were simplified, and then applied to a real case of half sibling relationship identification. At the same time, the LR of half sibling relationships under the assumption that only a single biological mother or none of the biological mothers participate were respectively calculated. Results In the cases of identification of half sibling relationship from same fathers, with no biological father involved, after the same genetic indicator test analysis, when both biological mothers participate in the identification, the accumulated LR value was higher than that of accumulated LR with only a single biological mother or no parents participating. Conclusion When the autosome STR test is used for the identification and analysis of half sibling relationship between two individuals, the calculation of LR is more simple, intuitive and operable with both mothers participating. The biological mothers should participate in the test as much as possible, otherwise the number of STR loci would need to be increased for a more specific conclusion.

Fluacrypyrim induced apoptosis in acute myeloid leukemia cell lines / 国际药学研究杂志

Objective To investigate the effect of fluacrypyrim on the induction of apoptosis in human acute myeloid leuke-mia(AML)cell lines,NB4,THP-1 and HL-60,and explore the related mechanisms.Methods Trypan blue dye exclusion assay was used to estimate the growth of NB4,THP-1,and HL-60 cells after treatment with various concentrations of fluacrypyrim(1.25, 2.5,5 and 7.5 μmol/L)for 72 h.Cell apoptosis was evaluated by AnnexinⅤ-FITC/PI double stainning for the NB4 and THP-1 cells treated with fluacrypyrim(1.25,2.5 and 5 μmol/L)for 48 h as well as the HL-60 cells treated with fluacrypyrim(2.5,5 and 7.5 μmol/L) for 72 h.Western blotting was used to examine the protein expression of apoptotic regulators Bax,Mcl-1 and Caspase 3 in the NB4 cells treated with fluacrypyrim(1.25,2.5 and 5 μmol/L)for 24 h.Then,NB4 Cells were pretreated with Caspases inhibitor benzyloxy-carbonyl-Val-Ala-Asp-fluoromethylketone(Z-VAD-FMK)and exposed to fluacrypyrim at 2.5 μmol/L for 24 h,which was then evaluat-ed for the apoptosis using AnnexinⅤ-FITC/PI double stainning.Western blotting was used to examine the expression of the phosphory-lated and total proteins of mitogen-activated protein kinase(MAPK)signaling molecules,ERK,JNK and P38,in the NB4 cells treat-ed with fluacrypyrim(1.25,2.5 and 5 μmol/L)for 24 h. NB4 Cells were pretreated with ERK inhibitor U0126,JNK inhibitor SP600125,or P38 inhibitor SB203580 for 1 h and then exposed to fluacrypyrim at 2.5 μmol/L for 24 h,which was then analyzed for the apoptosis by the AnnexinⅤ-FITC/PI double stainning.Results The proliferation of NB4,THP-1 and HL-60 cells was inhibited by the treatment with fluacrypyrim(2.5,5 and 7.5 μmol/L)for 72 h.The apoptosis induced in the NB4 and THP-1 cells by the fluacry-pyrim treatment at 5 μmol/L for 48 h and in the HL-60 cells by the fluacrypyrim treatment at 7.5 μmol/L for 72 h were significant as compared with the control group(P<0.01).Mechanically,fluacrypyrim at the concentrations of 2.5 and 5 μmol/L effectively up-regu-lated the expression of Bax(P<0.01 and P<0.05)for the 2.5 and 5 μmol/L,respectively,down-regulated the expression of Mcl-1 (P<0.01)and activated Caspase 3(P<0.01)in the NB4 cells when compared with the control group(P<0.01).The pretreatment of the NB4 cells with Z-VAD-FMK blocked the apoptosis induced by fluacrypyrim.Furthermore,the fluacrypyrim(2.5 and 5 μmol/L) treatment increased the ERK,JNK and P38 phosphorylation(P<0.01),while the pretreatment of the NB4 cells with U0126 signifi-cantly inhibited the the fluacrypyrim-induced apoptosis(P<0.01),as compared with the control group.Conclusion Fluacrypyrim effectively inhibits the cell proliferation and induces caspase-dependent cell apoptosis in AML cells.Activation of ERK/MAPK signal-ing pathway might play an important role in the action of fluacrypyrim.

Efficacy and Safety of Orlistat versus Metformin in Reducing Body Weight of Overweight or Obese Patients:A Meta-analysis / 中国药房

OBJECTIVE: To evaluate therapeutic efficacy and safety of orlistat versus metformin in reducing body weight of overweight or obese patients, and to provide evidence-based reference.METHODS: Retrieved from PubMed, Embase, Ovid, Web of Science, Cochrane Library, Chinese Journal Full-text Database, Wanfang database and VIP, RCTs about orlistat alone or combined with metformin (trial group) versus metformin alone (control group) in reducing body weight, BMI and the incidence of ADR of overweight or obese patients were collected. Meta-analysis was performed by using Rev Man 5. 3 statistical software after data extraction and quality evaluation with modified Jadad scale. RESULTS: A total of 9 RCTs were included, involving 502 patients. The results of Meta-analysis showed that, when orlistat combined with metformin, the reduction of BMI in trial group was significantly better than control group, with statistical significance [SMD= -0. 74, 95％CI (- 1. 22,-0. 26),P=0. 002]. There was no statistical significance in the reduction of body weight [SMD= -0. 04, 95％CI (-0. 27,0. 20), P=0. 76] or the incidence of ADR [RR=1. 07, 95％CI (0. 68, 1. 68), P=0. 78] between 2 groups. CONCLUSIONS: Both orlistat and metformin can reduce body weight with good safety. Combined use of these two drugs can reduce body weight more significantly.

Evaluation of iodixanol combined with low voltage in CT imaging quality of solitary pulmonary nodules / 实用医学杂志

Objective To explore the feasibility of low concentration contrast medium and low-voltage combined with adaptive statistical iterative reconstruction(ASRI)technique in enhanced CT imaging of solitary pulmonary nodules. Methods A total of 40 patients with solitary pulmonary nodules who underwent routine exam-inations and were pathologically confirmed from February 2015 to February 2017 were collected and divided into conventional group(conventional dose,high osmolar contrast,using filtered back projection reconstruction)and low dose group(low voltage,low concentration isotonic contrast,iterative reconstruction). Results Subjective scoring of conventional group(3.97 ± 0.57)and low dose group(4.01 ± 0.54)indicated no statistical significance (P > 0.05). No significant difference was found regarding to reconstructed image quality,SNR and CNR in both two groups.The dose length product(DLP)and effective dose(ED)in low dose group were lower than those in the conventional group[(283.52 ± 11.50)mGy/cm vs(370.74 ± 29.56)mGy/cm;(3.65 ± 0.32)mSV vs(5.11±0.25) mSV],and the difference was statistically significant(P < 0.05). Conclusions Low concentration isotonic con-trast agent(iodixanol 270 mgI/L)and low voltage(100 kV)combined with ASIR technology could satisfy the clini-cal need in enhanced CT imaging of solitary pulmonary nodule.

Mutations in a Large Pedigree with Y-STR Genetic Markers / 法医学杂志

OBJECTIVE@#To explore the mutation of Y-STR loci in meiotic allelic transmission in a large pedigree.@*METHODS@#The oral swabs of 163 male individuals were collected from a Lin pedigree. Twenty-two Y-STR genetic markers were typed with AGCU Y24 fluorescent detection kit (AGCU Y24 system), which also contained 16 Y-STR markers included in Yfiler multiple amplification kit (Yfiler system). The genotyping results of Y-STR loci were compared between each two males in the pedigree.@*RESULTS@#There were 20 and 30 kinds of haplotypes obtained with Yfiler and AGCU Y24 systems in 163 male individuals from the Lin pedigree, respectively. The rates referred to haplotype differences (RRHD) of these two typing systems between male pairs were 0.910 5 and 0.922 7, respectively. The average number of marker differences were 6.582 1 and 9.824 8, respectively. The RRHD increased along with the incidents of meiosis.@*CONCLUSION@#Y-STR mutation leads to different Y-STR haplotypes among the male members in a paternal pedigree and the rate of difference increases along with the incidents of meiosis.

Effect of GW003 on the granulocyte macrophage colony formation ability of bone marrow cells in vitro / 中国实验血液学杂志

The aim of this study was to investigate the effect of GW003 on the ability of granulocyte colony forming in vitro of bone marrow cells. The bone marrow samples was collected from normal rhesus, the patients with leukemia in stages of remission and chemotherapy respectively, and the nucleated cells were separated and cultured for 12 days after addition of different concentrations of GW003 or rhG-CSF, or G-CSF mutant. Then the amount of colony-forming unit-granulocyte-macrophage was counted. The results indicated that GW003 could enhance the ability of bone marrow nucleated cells of rhesus to forming CFU-GM in vitro, and its effect was much better than that of rhG-CSF or G-CSF mutant at the same concentration(®). The GW003 showed dose-response relationship to CFU-GM level (r = R(2) = 0.965, P = 0.003, in a certain concentration), the GW003 also could enhance CFU-GM formation of marrow nucleated cells in leukemic patients, especially for patients receiving chemotherapy. The GW003 could relieve the marrow suppression caused by chemotherapy significantly. It is concluded that the GW003 can significantly improve the ability of bone marrow cells to form granulocyte colony in vitro as well as effectively alleviate bone marrow suppression.

Effect of homeobox A2 gene RNA interference on HCC cell behavior / 安徽医科大学学报

Objective To study the role of homeobox A2(HOXA2) gene in cell growth,cellcycle and apoptosis of hepatoma cells, and discuss the feasibility that HOXA2 gene would be a potential therapy target of hepatoma. Methods Real-time quantitative polymerase chain reaction and revers transcriptional polymerase chain reaction were performed to examine HOXA2 expression in tumorous and non-tumorous tissue of patients was matched with liver cancer. siRNAs were chemically synthesized to interference HOXA2 in PLC/PRF/5 and MHCC-97L cells. Growth curve and soft-agar colony formation assay were performed to evaluate cell growth, and cell cycle and apop-tosis were analyzed by flow cytometry. Results HOXA2 was upregulated in HCC samples compared with matched non-tumor tissues(P<0.05). Two siRNAs against HOXA2 gene were designed and synthesized to specially knock-down HOXA2 in PLC/PRF/5 and MHCC-97L cells. HOXA2 knockdown inhibited cellular growth and soft-agar colony formation in PLC/PRF/5 and MHCC-97L cells. And Fowcytometry analysis revealed that HOXA2 knock-down by RNA interference could result in G1 arrest and S decrease and promoted cellular apoptosis. ConclusionHOXA2 gene has an important role in cell growth of hepatoma cells.

Age estimation using content of sjTREC in human peripheral blood / 法医学杂志

OBJECTIVE@#To determine and verify the correlation formula of age estimation using the content of signal joint T-cell receptor excision DNA circle (sjTREC) in human peripheral blood and to discuss its application value in forensic biological practice.@*METHODS@#The samples of peripheral blood stains were collected from 30 healthy unrelated individuals whose ages were known. The DNAs were extracted from the samples stored at room temperature after 4 weeks. The content of sjTREC was measured by real-time fluorescent quantitative PCR technique, and the TATA box binding protein (TBP) was selected as reference genes. The age of each sample was predicted with the formula which was Age = -7.181 5 Y-42.458 +/- 9.42 (Y = dCtTBP-sjTREC), and the result was compared with the real age of each individual to determine the accuracy of the formula.@*RESULTS@#sjTREC and TBP gene were detectable in all 30 samples of peripheral blood. The contents of sjTREC in human peripheral blood showed a decreasing tendency with aging. The accuracy rate for the age estimation by this method was 76.67%.@*CONCLUSION@#The method for the age estimation with the content of sjTREC was simple, fast, sensitive, and good species specific with important potential application prospect.

Construction and expression of recombinant adenovirus containing human catalase gene in vitro / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To construct the adenovirus vector containing recombinant human catalase (CAT) and to express the recombinant gene in vitro.</p><p><b>METHODS</b>Total RNA was extracted from human leukocytes and full-length human CAT cDNA was obtained with RT-PCR method. The CAT gene was cloned into pcDNA3.1(+) vector and pcDNA3.1(+)CAT was constructed. The positive clones were confirmed by the restriction enzyme digestion and gene sequencing. The CAT gene was cloned into the entry vector pENTR1A, and pENTR1A-CAT vector was constructed. By LR reaction pENTR1A-CAT and pAd/CMV/V5-DEST was recombined in vitro, and the recombinant adenovirus pAd/CMV/V5-DEST-CAT was obtained. The positive pAd/CMV/V5-DEST-CAT was confirmed by sequencing and transfected into 293A cells with Pac I linearization and Lipofectamine 2 000, and the recombinant virus particles were packaged and amplified in the cells. The expression of CAT protein and CAT enzyme activities of the recombinant virus were determined by Western blot and 240 nm UV absorption methods.</p><p><b>RESULT</b>High expression of recombinant adenovirus was obtained and the expressed human catalase had high enzyme activity.</p><p><b>CONCLUSION</b>Ad/CMV/V5-DEST-CAT vector containing human catalase gene has been constructed successfully; and the expressed enzyme in 293A cells has high activity.</p>

High dose granulocyte colony-stimulating factor enhances survival and hematopoietic reconstruction in canines irradiated by 2.3 Gy mixed fission neutron and gamma ray / 中国实验血液学杂志

This study was purposed to evaluate the effects of recombinant human granulocyte colony-stimulating factor (rhG-CSF) on hematopoietic reconstruction and survival in beagles exposed to mixed fission neutron and γ-ray. 13 beagles were unilaterally exposed to single dose of 2.3 Gy 90% neutrons. The experiments were divided into 3 groups: irradiation control group (no any treatment, n = 4), supportive care group (n = 5) and rhG-CSF plus supportive care group (n = 4, abbreviated as rhG-CSF group) in which the beagles were subcutaneously injected with 200 µg/kg of rhG-CSF early at half an hour and 24 hours post-irradiation respectively. The results showed that 2.3 Gy 90% neutron irradiation induced a severe acute radiation sickness of bone marrow type. The administration of rhG-CSF increased the survival rate from 60% in supportive care group to 100%. Twice injection of rhG-CSF in the first 24 hours reduced duration of neutropenia, enhanced neutrophil nadir and promoted neutrophil recovery when compared with control cohort administered clinical support. The number of colony-forming cells (CFU-GM, CFU-E, and BFU-E) in peripheral blood of rhG-CSF treated canines increased 2-to 5-fold relative to those of the supportive care group on day 3. All canines treated with rhG-CSF achieved hematopoietic reconstruction as evidenced by the pathological section of sternum while severe shortage of hemopoietic cells remained in the cohorts given supportive care alone. It is concluded that the combination of supportive care and high-dose rhG-CSF can accelerate hematopoietic recovery and enhance survival of dogs exposed to 2.3 Gy mixed neutron and gamma ray.

Calculation of the avuncular index / 法医学杂志

OBJECTIVE@#To introduce the method of avuncular index (AI) calculation.@*METHODS@#Identity by decent coefficient, coancestry coefficient and AI law were employed in identification of uncle-niece relationship, when autosomal STR loci were detected to determine controversial uncle-niece relationship.@*RESULTS@#The results of AI calculation were coincidental using identity by descent coefficien, coancestry coefficient and AI law.@*CONCLUSION@#The results are coincidental using three methods in the different situations. AI index is higher with participation of children's mother.

The infection of dendritic cells by recombinant adenoviral vector carrying HBsAg-HSP70 chimeric gene and its biological characteristics observations / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To test the infeciton efficiency of recombinant adenoviral vector carrying HBsAg-HSP70 chimeric gene and to abserve its biological characteristics.</p><p><b>METHODS</b>Peripheral blood mononuclear cells (PBMC) were separated from healthy blood donor and they were infected by Ad-HSP70-HBsAg on the first day of isolation. DCs were induced in medium with cytokines IL-4, GM-CSF and TNF-alpha in vitro. The biological characteristics of DC induced were analyzed by inverted fluorecent microscope, RT-PCR, flow cytometer (FACS), and mixed lymphocyte reaction (MLR).</p><p><b>RESULTS</b>The traced gene-GFP were abserved in DCs by inverted fluorecent microscope and HSP70-HBsAg gene mRNA expression was detected by RT-PCR after the Ad-HSP70-HBsAg infection. FACS analysis shown that the expression of CD1a, CD80, CD86 and HLA-DR on surfece of two groups of DCs were similar. MLR showed that there are not a statitic difference of stimulated index (SI) between two groups.</p><p><b>CONCLUSION</b>Results indicated that Ad-HSP70-HBsAg can effectively infected DCs without affecting its biological characteristics.</p>

Cloning of Rcet3, a novel gene related to family 2 cystatins / 南方医科大学学报

<p><b>OBJECTIVE</b>To clone the full-length Rcet3 gene, a novel gene related to family 2 cystatins, from mouse testis or other tissues.</p><p><b>METHODS</b>Rcet3 gene was cloned using digital differential display (DDD) and RT-PCR was performed for cloning the full-length Rcet3 gene from adult mouse testis cDNA library with sequence analysis.</p><p><b>RESULTS</b>Rcet3 cDNA was 610 bp in length, consisting of 4 exons to encode a protein with 140 amino acid residues. The encoded protein contained a potential signal peptide and a cystatin domain, but lacked critical consensus site important for cysteine protease inhibition. These characteristics could be seen in the Cres subgroup related to the family 2 cystatins. Rcet3 was specifically expressed in adult mouse testis, epididymis and the cerebrum, but at higher levels in the testis than in the epididymis and cerebrum.</p><p><b>CONCLUSION</b>Rcet3 may be a new member of Cres subgroup of family 2 cystatins.</p>

Expression of fusion gene PAX3/PAX7-FKHR and chromosomal aberration in rhabdomyosarcoma / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To detect the PAX3/PAX7-FKHR fusion transcripts to identify genetic alteration in embryonal rhabdomyosarcoma (ERMS) and alveolar rhabdomyosarcoma (ARMS) tissues.</p><p><b>METHODS</b>One-step reverse transcription- polymerase chain reaction (RT-PCR) were used to detect the expression of the PAX3/PAX7-FKHR fusion transcrips in 16 cases of rhabdomyosarcoma (7 cases of ARMS, 9 cases of ERMS) and 16 specimens were compared to the surrounding normal tissue. Comparative genomic hybridization (CGH) was employed to detect the genomic imbalance (DNA loss or amplification) in 16 RMS cases.</p><p><b>RESULTS</b>PAX3-FKHR fusion transcripts were positive in 3/7 and PAX 7-FKHR fusion transcripts were positive in 2/7 of ARMS patients, respectively, and were all negative in ERMS and Control tumors. There were different chromosome variations for each RMS, chromosome amplification was frequently seen in 1p36 (69%), 5q32 (56%), 8q21 (63%), 13q14 (69%), 19q (63%), 20q (56%). Chromosome loss was frequently seen in 3p21-pter (56%), 9p23-pter (50%), 10q (69%), 16/16q24 (56%).</p><p><b>CONCLUSION</b>One-step RT-PCR assay for detection specific fusion gene provides a useful tool for confirmation of the diagnosis of RMS in diagnostically difficult cases and in retrospective studies. Chimeric gene transcript resulting from specific chromosomal translocations is a reliable index for the molecular diagnosis of RMS.</p>

Genomic imbalance and chromosome disorders in hepatoblastoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To establish stable techniques of comparative genomic hybridization (CGH) and apply them to elucidate the genetic characteristics of hepatoblastoma (HB), and to explore the characteristics and clinical significance of loss of heterozygosity (LOH) at 1p36 in HB.</p><p><b>METHODS</b>CGH was employed to detect the genomic imbalance (DNA loss or amplification) in 20 cases of HB, and PCR-simple repeated sequence polymorphism was employed in 30 cases of HB to detect the loss of heterozygosity for 6 satellites at chromosome 1p36.</p><p><b>RESULTS</b>There were different chromosome variations for each HB. chromosome amplification was frequently seen in 1q, 2q,2p, 8q, 8p, 12q and 22q. Chromosome loss was often seen in 1p, 4q, 4p, 16q, 17p and 18q. The frequency of LOH at 6 loci on chromosome 1 was 63.3% totally (19/30), with the highest D1S199 (66.7%) and D1S450 next to it (46.7%).</p><p><b>CONCLUSION</b>There were chromosome zones with DNA amplification or loss in hepatoblastoma. There are extensive LOH at 1p36 in hepatoblastoma. The corresponding amplification of oncogene and loss of antioncogene may take part in the development of hepatoblastoma.</p>