[Specifications for diagnosis and treatment of non-neonatal tetanus].

Tetanus consists of neonatal tetanus and non-neonatal tetanus. Non-neonatal tetanus remains a serious public health problem, although neonatal tetanus has been eliminated in China since 2012. Non-neonatal tetanus is a potential fatal disease. In the absence of medical intervention, the mortality rate of severe cases is almost 100%. Even with vigorous treatment, the mortality rate is still 30%-50% globally. These specifications aim to regulate non-neonatal tetanus diagnosis and treatment in China, in order to improve medical quality and safety. These specifications introduce the etiology, epidemiology, pathogenesis, clinical manifestations and laboratory tests, diagnosis, differential diagnosis, grading and treatment of non-neonatal tetanus.

[Specifications for diagnosis and treatment of non-neonatal tetanus].

Tetanus consists of neonatal tetanus and non-neonatal tetanus. Although neonatal tetanus in China has been eliminated since 2012, non-neonatal tetanus remains a serious public health problem. Non-neonatal tetanus is a potential fatal disease, and the mortality rate of severe cases is almost 100% in the absence of medical intervention. Even with vigorous treatment, the mortality rate is still 30~50% globally. In order to standardize the diagnosis and treatment of non-neonatal tetanus in China, this specification is hereby formulated. This standard includes etiology, epidemiology, pathogenesis, clinical manifestations, laboratory tests, diagnosis, differential diagnosis, classification, grading and treatment of non-neonatal tetanus.

Targeting Mdmx to treat breast cancers with wild-type p53.

The function of the tumor suppressor p53 is universally compromised in cancers. It is the most frequently mutated gene in human cancers (reviewed). In cases where p53 is not mutated, alternative regulatory pathways inactivate its tumor suppressive functions. This is primarily achieved through elevation in the expression of the key inhibitors of p53: Mdm2 or Mdmx (also called Mdm4) (reviewed). In breast cancer (BrCa), the frequency of p53 mutations varies markedly between the different subtypes, with basal-like BrCas bearing a high frequency of p53 mutations, whereas luminal BrCas generally express wild-type (wt) p53. Here we show that Mdmx is unexpectedly highly expressed in normal breast epithelial cells and its expression is further elevated in most luminal BrCas, whereas p53 expression is generally low, consistent with wt p53 status. Inducible knockdown (KD) of Mdmx in luminal BrCa MCF-7 cells impedes the growth of these cells in culture, in a p53-dependent manner. Importantly, KD of Mdmx in orthotopic xenograft transplants resulted in growth inhibition associated with prolonged survival, both in a preventative model and also in a treatment model. Growth impediment in response to Mdmx KD was associated with cellular senescence. The growth inhibitory capacity of Mdmx KD was recapitulated in an additional luminal BrCa cell line MPE600, which expresses wt p53. Further, the growth inhibitory capacity of Mdmx KD was also demonstrated in the wt p53 basal-like cell line SKBR7 line. These results identify Mdmx growth dependency in wt p53 expressing BrCas, across a range of subtypes. Based on our findings, we propose that Mdmx targeting is an attractive strategy for treating BrCas harboring wt p53.

Human leukocyte antigen class II alleles in Caucasian women with primary biliary cirrhosis.

In the current study, we investigated human leukocyte antigen (HLA) class II alleles in Caucasian women with primary biliary cirrhosis (PBC), a disease that preferentially affects women. Alleles of DRB1, DQA1, and DQB1 were determined by DNA-based HLA typing for women with PBC (n = 72) and healthy women (n = 381). All study subjects were Caucasian. HLA DRB1*08 was significantly increased in women with PBC compared to healthy women. The increase was primarily due to the DRB1*0801 allele, also the most common DRB1*08 allele among controls. DQB1*04 and DQA1*0401 were significantly increased. DRB1*1501, DQA1*0102, and DQB1*0602 were associated with decreased risk. Analyses conducted comparing parous women with PBC to parous healthy women (n = 68 and n = 282, respectively) yielded similar significant results. Although the DRB1*08-DQA1*0401-DQB1*04 haplotype was significantly associated with PBC, consistent with other studies, this haplotype nevertheless represented only 19% (14/72) of all PBC patients and can account for only a minority of the risk of PBC.

Male microchimerism in women with systemic sclerosis and healthy women who have never given birth to a son.

BACKGROUND: Male DNA or cells are often used to measure microchimerism in a woman. In studies of autoimmune diseases male microchimerism is most often attributed to the previous birth of a son. OBJECTIVE: To determine the frequency of male microchimerism in healthy women or women with systemic sclerosis who had never given birth to a son. METHODS: Real time quantitative polymerase chain reaction targeting the Y chromosome specific sequence DYS14 was employed to test DNA extracted from peripheral blood mononuclear cells of 26 women with systemic sclerosis and 23 healthy women who had never given birth to a son. RESULTS: are expressed as the genome equivalent number of male cells per million host cells (gEq/mil).Results: Male DNA was found in 15% of women with systemic sclerosis (range 0 to 23.7 gEq/mil) and in 13% of healthy women (range 0 to 5.1 gEq/mil). Although two women with male DNA had an induced abortion, most had no history of spontaneous or induced abortion (either systemic sclerosis or healthy). CONCLUSIONS: Microchimerism with male DNA can be found in the circulation of women who have never given birth to a son. Thus sources other than a male birth must be considered when male DNA is used to measure microchimerism. Although other studies are needed, there was no apparent difference in women with systemic sclerosis and healthy women. Possible sources of male DNA include unrecognised male pregnancy or unrecognised male twin, an older male sibling with transfer through the maternal circulation, or sexual intercourse alone.

HLA allelic variants encoding DR11 in diffuse and limited systemic sclerosis in Caucasian women.

OBJECTIVE: We investigated HLA class II alleles in women with systemic sclerosis (SSc), a rare disease that preferentially affects women. METHODS: Specific alleles of DRB1, DQA1 and DQB1 were determined by DNA-based HLA typing for women with SSc (n = 102) and healthy women (n = 533). All study subjects were Caucasian. DRB1, DQA1 and DQB1 allele frequencies of women with SSc were compared with those of healthy women. RESULTS: Among women with SSc, 29.4% (30/102) and among healthy women 10.7% (57/533) had DRB1*11. Allele frequencies were compared for women with SSc and healthy women (each woman has two alleles). The allele frequency of DRB1*11 was 15.7% (32/204 alleles) in SSc women and 5.8% (62/1066 alleles) in healthy women (P = 0.000002). The increase of DRB1*11 was found both in diffuse (P = 0.0001) and limited SSc (P = 0.002) (allele frequencies 15.0 and 17.2%, respectively). Among women with diffuse SSc, there was a disproportionate increase of the DRB1*1104 allele (P = 0.0004) with no increase of DRB1*1101 (P = 1.00). In contrast, in limited SSc the strongest association was with DRB1*1101 (P = 0.008), with a less significant increase of DRB1*1104 (P = 0.04). CONCLUSIONS: An increase of DRB1*11 in SSc is consistent with other reports. Although present in both diffuse and limited SSc disease subsets, the increase was predominantly due to over-representation of DRB1*1104 in women with diffuse SSc. Women with limited SSc had a preponderance of DRB1*1101, the most common allele in healthy women. DRB1*1104 and DRB1*1101 differ by a single amino acid at position 86, where the former has valine and the latter glycine.

Toxicity of allyl alcohol in primary cultures of freshly isolated and cryopreserved hepatocytes maintained on hydrated collagen gels.

The toxicity of allyl alcohol was compared in freshly isolated and cryopreserved hepatocytes that were either placed in suspension or maintained on hydrated collagen gels in a sandwich configuration. The purpose of this study was to evaluate whether the two types of cells displayed the same sensitivity to allyl alcohol when maintained in vitro over relatively prolonged periods of time. The important differentiated functions of urea synthesis, secretion of albumin, and metabolism of ethoxycoumarin, a model drug substrate, were used as end points of toxicity. Cryopreserved hepatocytes incubated in physiological buffer shortly after removal from liquid nitrogen were more sensitive to allyl alcohol than freshly isolated hepatocytes. In contrast, cryopreserved and freshly isolated hepatocytes maintained on hydrated collagen gels responded identically to allyl alcohol. Thus, the increased sensitivity of cryopreserved hepatocytes in suspension to allyl alcohol is a transient phenomenon that disappears after the cells have been allowed to recover on hydrated collagen gels. Dissipation of the mitochondrial membrane potential by allyl alcohol, as indexed by rhodamine 123 fluorescence, was also the same in freshly isolated and cryopreserved hepatocytes maintained on hydrated collagen matrices. This loss of mitochondrial membrane potential caused by allyl alcohol preceded inhibition of albumin and urea biosynthesis. Collectively, the results indicate that cryopreserved cells maintained on hydrated collagen gels provide a useful system to define the actions of certain hepatotoxic agents over relatively prolonged periods of time in vitro.

Actions of selected hepatotoxicants on freshly isolated and cryopreserved rat hepatocytes.

The present study compares the actions of the hepatotoxic agents allyl alcohol, acetaminophen, and carbon tetrachloride on energy metabolism in freshly isolated and cryopreserved rat hepatocytes. After 30 min incubation of freshly isolated hepatocytes at 37 degrees C to allow metabolic equilibration, hepatocytes were supplemented with cryoprotectants and cooled in a stepwise manner to liquid nitrogen temperature. Hepatocytes stored in liquid nitrogen for 2 weeks to 6 months were thawed and centrifuged through Percoll to remove damaged cells. Despite similarities in energy status as indexed by ATP content and the rate of urea synthesis in freshly isolated and cryopreserved hepatocytes, cryopreserved hepatocytes were more sensitive to hepatotoxicants. All three hepatotoxicants caused ATP and rates of urea synthesis to decline to a greater extent in cryopreserved than in freshly isolated hepatocytes. Rates of oxygen uptake were higher in cryopreserved cells than in freshly isolated hepatocytes and declined in cryopreserved cells but not in freshly isolated cells during the initial period of incubation. Rates of mitochondrial respiration stimulated with site-specific substrates were comparable in freshly isolated and cryopreserved cells permeabilized with digitonin. Allyl alcohol and acetaminophen inhibited site-specific respiration to the same extent in both groups of cells. Collectively, these results suggest that increased sensitivity to hepatotoxic agents and elevated oxygen consumption in cryopreserved hepatocytes recovered after storage in liquid nitrogen are related to higher demand for energy in these cells rather than to permanent injury caused by cryopreservation and irreversible uncoupling of oxidative phosphorylation.