Strengthening medical ethics education in clinical practice of internal medicine / 上海预防医学

With rapid society development and the constant changes in people's thinking， the medical ethics issue has become more prominent. Medical interns are often facing ethical problems in clinical practice. Due to the lack of understanding of the experience in medical ethics， they often handle the problems poorly， which leads to intensified disputes between doctors and patients. The paper discusses some ethical problems that often appear in the clinical practice of internal medicine， and suggests several methods to improve the medical ethics education of interns in the clinical process.

Effect and Mechanism of Traditional Chinese Medicine on Prevention and Treatment of Pathological Scar / 中国实验方剂学杂志

Pathological scar is a kind of skin fibrotic disease caused by abnormal wound healing, including hypertrophic scar and keloid. Pathological scar may lead to aesthetic flaws, limb dysfunction and local discomfort in patients. Due to the complexity of the wound healing process, the formation of scar is affected by many factors. In addition to traditional surgical, laser, cryostatic and hormone injection methods for the treatment of pathological scar, there are new therapies, such as mesenchymal stem cell therapy, fat transplantation, interferon, and botulinum toxin. They are widely used in clinical practice, but with such problems as high prices and many side effect. Traditional Chinese medicine (TCM) has a long history in treating pathological scar. In recent years, in vivo and in vitro studies have shown that TCM has effect IN reducing inflammation, inhibiting fibroblast proliferation, regulating fibroblast activation and migration, inducing fibroblast apoptosis and autophagy, promoting the degradation of extracellular matrix (ECM) and reducing angiogenesis in general. Besides, TCM has also a certain regulatory role in the signaling pathways, such as transforming growth factor-β1 (TGF-β1)/Smads, phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin (PI3K/Akt/mTOR) and sonic hedgehog (Shh). There are still some contradictions in relevant studies, and specific mechanisms remain to be further improved. This paper summarizes the study content, findings and relevant mechanisms of different TCM based on in vivo and in vitro experiments, analyzes the advantages and disadvantages of TCM in the prevention and treatment of pathological scar, and its prospects in clinical application, so as to provide basis and ideas for future scar studies.

Effect factors and mechanisms of borneol's bidirectional regulation on blood-brain barrier permeability / 中国中药杂志

In recent twenty years, there are a lot of studies about the effect of borneol on permeability of blood-brain barrier(BBB); however, it isDODOrt of regular conclusions of effect factors and in-depth analysis of functional mechanisms. The current researching data were collected and analyzed in this paper for illuminating the effect factors and mechanisms of borneol on permeability of BBB.The following conclusions were obtained: five factors about borneol influencing the permeability of BBB. First, opticity activity of borneol had no significant effect on action effects. Second, dose of borneol in the range of 50.00-200.00 mg•kg⁻¹, did not affect the effect direction, but only affect its action intensity either with use alone or combination use. Third, the borneol can increase the permeability of physiological BBB, and decrease the permeability of pathological BBB. Fourth, regardless of using singly or using compatibility with musk, borneol can decrease the permeability of BBB in different brain disease models. Fifth, when used with astragalus, catalpol or puerarin, borneol can increase the permeability of BBB and promote the drugs through BBB in pathological conditions. The target spots and mechanisms of borneol's bidirectional regulation on the permeability of BBB are related to the structure and function of cerebral endothelial cells, the exocytosis effects of P-gp and low pinocytosis internal transport effects. On one hand，borneol can down-regulate P-gp by inhibiting NF-κB to reduce the exocytosis effects of P-gp and promote the blood brain barrier pinocytosis to increase the permeability of BBB; On the other hand，borneol can reduce the degradation of basement membrane of blood vessel and tight junctions by inhibiting the expression of IL-1β, MMP-9 to decrease the permeability of BBB；moreover，borneol has bidirectional regulation effects on blood-brain barrier permeability by influencing the signaling pathways of Ca2+-eNOS-NO, VEGF-eNOS-NO. However, the detailed mechanisms that borneol regulates and controls the permeability of BBB are so complicated, so they shall be further proved and clarified.

Borneol promotes catalpol and puerarin crossing through blood-brain barrier in focal cerebral ischemic rats / 中国中药杂志

Previous studies showed that borneol could promote some drugs crossing through the blood-brain-barrier (BBB) at certain conditions. However, the mechanism has not been clarified yet. This study aimed to investigate the effect of bornrol on promoting catalpol and puerarin through BBB and explore the relevant mechanism. The focal cerebral ischemic rats were divided into 7 groups randomly and then were administered corresponding drugs: model group (M, solvent), catalpol-puerarin group (ZG, catalpol 45 mg•kg⁻¹+puerarin 200 mg•kg⁻¹), catalpol-puerarin-bornrol group(ZGB, catalpol 45 mg•kg⁻¹+puerarin 200 mg•kg⁻¹ +bornrol 200 mg•kg⁻¹), catalpol-bornrol group(ZB, catalpol 45 mg•kg⁻¹ +bornrol 200 mg•kg⁻¹), puerarin-bornrol group(GB, puerarin 200 mg•kg⁻¹ +bornrol 200 mg•kg⁻¹), butoxamine-ZG group(BTX+ZG, butoxamine 1.5 mg•kg⁻¹+ catalpol 45 mg•kg⁻¹+puerarin 200 mg•kg⁻¹), and butoxamine-ZGB group(BTX+ZGB, butoxamine 1.5 mg•kg⁻¹+ catalpol 45 mg•kg⁻¹+puerarin 200 mg•kg⁻¹ +bornrol 200 mg•kg⁻¹). Another 10 sham-operated rats were set as control (S). Ten minutes after the administration, the cerebrospinal fluid was taken to test the content of catalpol and puerarin, and the brain tissue was taken to test the expression of β2-adrenergic receptor, eNOS, and NO. Compared with the M group, the ZG group showed content of catalpol is 26.673 μg•L⁻¹ and the content of puerarin is below the detection limit;the expression levels of β2-adrenergic receptor, eNOS and the contents of NO in brain tissue are no significant difference. Compared with the ZG group, the ZGB, ZB and GB groups showed significantly increased content of catalpol andpuerarin, as well as the expression of β2-adrenergic receptor, eNOS and NO in the brain tissue (P<0.05). The content of catalpol in BTX+ZG group changed non-significantly. Compared with the ZGB group, the BTX+ZGB group presented significantly decreased content of catalpol and puerarin and reduced expression of eNOS and NO in the brain tissue (P<0.05）.The results demonstrated that borneol could dramatically promote catalpol and puerarin crossing through BBB in the focal cerebral ischemic rats. Moreover, the effect may be related to the up-regulation of β2-adrenergic receptor and the increasing expression of eNOS and NO.

Molecular basis of partial D phenotypes in Chinese / 中国实验血液学杂志

To investigate the molecular basis of partial D phenotypes in Chinese, D variants with weak D expression was screened by using indirect anti-human globulin test (IAT) method, the polymerase chain reaction-sequence specific primer (PCR-SSP) method was employed to amplify RHD specific exons and their flanking regions. The amplification products were sequenced directly to determine the molecular basis of D variants. The results showed that ten cases of partial D phenotypes, including one case of D Va (Kou.), one case of D Va (Hus.), one case of D Va-like (YH.), and seven cases of D VI type III, were detected from 22 cases of weak D phenotype respectively. All ten cases of partial D phenotypes had one RHD allele deleted. In conclusion, the molecular basis of ten cases of partial D phenotype was confirmed, including D Va (Kou.) and D Va-like (YH.) phenotypes reported firstly in Chinese population.

FUT3 gene polymorphism associated with Lewis blood group in Chinese Zhejiang population / 中国实验血液学杂志

To investigate the alpha-1, 3/4-fucosyltransferase gene (FUT3) polymorphism associated with Lewis blood group in Zhejiang population, the Lewis phenotypes of 183 random samples from Chinese blood donors in Zhejiang province were identified by standard serological techniques. The entire coding region of FUT3 gene were amplified by PCR from genomic DNA of 39 Lewis negative and 9 Lewis positive phenotype samples and sequenced directly. The haplotypes of FUT3 allele were identified by TOPO cloning sequencing method. The results showed that the frequency of true Le (a-b-) phenotype in Zhejiang population was 10.4% according to serological and molecular biological methods. Five nucleotide acid variant sites (59T > G, 202T > C, 314C > T, 508G > A and 1067T > A) were detected in all 48 sequencing samples. Besides the wild type Le allele, 2 common (le(59, 1067) and le(59, 508) and 3 rare non-functional le alleles (le(59), le(1067) and le(202, 314) were found in this population. In conclusion, the polymorphism of non-functional FUT3 allele was found to be relatively variable in Chinese Zhejiang population.

Variants 467C > T and 539G > C of the alpha-1,3-N-acetylgalactosaminyltransferase allele responsible for A2 subgroup / 中国实验血液学杂志

The purpose of this study was to investigate the molecular genetic basis of A2 subgroup and identify the novel alleles at ABO locus in Chinese Han population. All seven exons and their flanking sequences, enhancer and promoter in the ABO gene of five samples from individuals with serological discrepancies were amplified by polymerase chain reaction (PCR); the PCR products were screened by directly sequencing; the haplotypes of exon 6 and 7 were analyzed by TOPO cloning sequencing. The results showed that five samples were identified as A2 or A2B subgroup by serological technology. The A201 and A205 alleles were confirmed in one A2B individual and one A2 individual, respectively. A novel A2 variant allele was identified in three A2B individuals. The two nucleotide acid alterations (467C > T and 539G > C) at the exon 7 resulting in two amino acid substitutions (P156L and R180P) in this novel allele were observed, when compared with A101 allele. It is concluded that the polymorphism of A2 allele is found to be relatively variable in Chinese population, and a novel A208 allele responsible for A2 subgroup is firstly reported.

Establishment of genotyping method for DEL phenotype in Zhejiang Han population / 中国实验血液学杂志

In order to establish a genotyping method for DEL phenotype in Zhejiang Han population, an AS-PCR method was developed according to the RHD 1227A allele sequence. Its specificity and sensitivity were assessed in two Rh negative populations whose RHD 1227A or DEL phenotype status was known. The results showed that in evaluation of the method by detecting 50 RHD 1227A positive and 50 RHD 1227A negative individuals, the genotyping method displayed a sensitivity of 100% and a specificity of 100%; in evaluation of the method by detecting 33 DEL positive and 89 DEL negative individuals, the sensitivity was 100%, however, there were two serologically negative samples which were confirmed as positive using genotyping method. After re-testing these two samples with serological method and sequence analysis, it was found that original serological method gave false negative results and genotyping method still showed 100% specificity. The minimal target DNA concentration of this genotyping method is 8.13 ng/microl. In conclusion, designed genotyping method can be used to identify DEL phenotype efficiently in Zhejiang Han Rh negative population.

Molecular basis of Rh DEL phenotype in Zhejiang Han population / 中国实验血液学杂志

This study was purposed to investigate the molecular basis of Rh DEL phenotype. Rh DEL phenotypes were identified by a serologic adsorption-elution method, the nucleotide sequences of ten RHD exons and exon-intron boundary regions were evaluated by a RHD gene-specific PCR-SSP (PCR-SSP, polymerase chain reaction-sequence specific primer) and sequencing. The results showed that out of 122 random Rh negative donors 35 Rh DEL phenotypes were identified through serologic method, including 6 RhCCdee (17.14%), 28 RhCcdee (80.00%), and 1RhCcdEe (2.86%). Sequence analysis indicated that all DEL phenotypes harbored a RHD 1227 G > A mutation in exon 9. D zygosity test revealed that 29 DEL phenotypes (28 RhCcdee and 1 RhCcdEe) had one RHD gene deleted, and 6 DEL phenotypes (6 RhCCdee) had homogenous RHD gene. It is concluded that RHD 1227A is an important genetic marker for Rh DEL phenotype in Zhejiang Han population.

RHD 1227A allele frequency among Rh negative population and random population / 中国实验血液学杂志

To investigate the frequency of RHD 1227A allele in Rh negative population and random population, an AS-PCR (allele specific-polymerase chain reaction) method was employed to detect RHD 1227A allele. RHD gene copy was determined by D zygosity test and RHD exon 9 nucleotide sequence analysis. The results showed that among 143 Rh negative donors, forty-one RHD 1227A allele carriers were detected, and 8 (19.51%) out of which were RhCCdee, 32 (78.05%) were RhCcdee, and 1 (2.44%) was RhCcdEe. Thirty-five Rh negative RHD 1227A carriers had RHD gene deletion, and the remaining carriers were RHD 1227A homozygous. Seven (1.43%) individuals were detected with RHD 1227A allele among 489 random donors. They were all G/A heterozygous at RHD 1227 site. Serological test indicated that they were normal Rh positive phenotype. It is concluded that the frequency of RHD 1227A allele is 16.43% among Rh negative population and 0.72% among the random population.

278C > T variant of the alpha-1, 3-galactosyltransferase allele responsible for Bw subgroup / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the molecular genetic basis of the Bw variant and identify novel alleles at ABO locus in Chinese Han population.</p><p><b>METHODS</b>Serological techniques were performed to characterize erythrocyte phenotype of a proband. Mutations of the ABO gene were screened by polymerase chain reaction, reverse transcription-polymerase chain reaction and DNA sequencing.</p><p><b>RESULTS</b>The proband was identified as Bw phenotype by serological technology and family study. A novel Bw variant allele was identified in the gDNA and cDNA. The novel allele was observed a missense mutation (278 C to T) at the exon 6 which resulted in an amino acid substitution (P93L) compared with B101 allele. The 278 C to T was the first report mutation position in exon 6 among Bw alleles, so the P93L amino acid substitution was different from others Bw variants which had amino acid substitutions in a conserved functional domain reported previously.</p><p><b>CONCLUSION</b>A novel Bw allele (278 C to T) responsible for Bw variant is reported in Chinese population.</p>

Study on the frequency of alpha-1,2-fucosyltransferase gene h4 allele (C35T) in Chinese population / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the h4 allele (C35T) frequency of alpha-1,2-fucosyltransferase gene in Chinese population.</p><p><b>METHODS</b>The polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method for identifying C35T variant was established by using PCR to amplify a 125 bp FUT1 gene fragment, including C35T variant sequence, and the PCR product was digested by Hae III restriction enzyme. One hundred and fifty-eight random samples from Chinese blood donor were screened by PCR-RFLP.</p><p><b>RESULTS</b>Among 158 Chinese individuals with normal ABO and H blood group phenotypes, 8 and 83 were homozygous with 35T/T and 35C/C, respectively, while 67 were heterozygous with 35C/T. The allele frequencies were compatible with Hardy-Weinberg equilibrium.</p><p><b>CONCLUSION</b>The C35T substitution of FUT1 gene is not a mutation which gives rise to a non-functional h allele responsible for para-Bombay phenotype but a single nucleotide polymorphism in Chinese population.</p>

Study on the molecular genetics basis for one para-Bombay phenotype / 中国实验血液学杂志

To investigate the molecular genetics basis for one para-Bombay phenotype, the red blood cell phenotype of the proband was characterized by standard serological techniques. Exon 6 and 7 of ABO gene, the entire coding region of FUT1 gene and FUT2 gene were amplified by polymerase chain reaction from genomic DNA of the proband respectively. The PCR products were purified by agarose gels and directly sequenced. The PCR-SSP and genescan were performed to confirm the mutations detected by sequencing. The results showed that the proband ABO genotype was A(102)A(102). Two heterozygous mutations of FUT1 gene, an A to G transition at position 682 and AG deletion at position 547-552 were detected in the proband. A682G could cause transition of Met-->Val at amino acid position 228, AG deletion at position 547-552 caused a reading frame shift and a premature stop codon. The FUT2 genotype was heterozygous for a functional allele Se(357) and a weakly functional allele Se(357), 385 (T/T homozygous at position 357 and A/T heterozygous at 385 position). It is concluded that the compound heterozygous mutation--a novel A682G missense mutation and a 547-552 del AG is the molecular mechanism of this para-Bombay phenotype.