Molecular Subtyping of Triple-Negative Breast Cancers by Immunohistochemistry: Molecular Basis and Clinical Relevance.

BACKGROUND: Molecular subtyping of triple-negative breast cancers (TNBCs) via gene expression profiling is essential for understanding the molecular essence of this heterogeneous disease and for guiding individualized treatment. We aim to devise a clinically practical method based on immunohistochemistry (IHC) for the molecular subtyping of TNBCs. MATERIALS AND METHODS: By analyzing the RNA sequencing data on TNBCs from Fudan University Shanghai Cancer Center (FUSCC) (n = 360) and The Cancer Genome Atlas data set (n = 158), we determined markers that can identify specific molecular subtypes. We performed immunohistochemical staining on tumor sections of 210 TNBCs from FUSCC, established an IHC-based classifier, and applied it to another two cohorts (n = 183 and 214). RESULTS: We selected androgen receptor (AR), CD8, FOXC1, and DCLK1 as immunohistochemical markers and classified TNBCs into five subtypes based on the staining results: (a) IHC-based luminal androgen receptor (IHC-LAR; AR-positive [+]), (b) IHC-based immunomodulatory (IHC-IM; AR-negative [-], CD8+), (c) IHC-based basal-like immune-suppressed (IHC-BLIS; AR-, CD8-, FOXC1+), (d) IHC-based mesenchymal (IHC-MES; AR-, CD8-, FOXC1-, DCLK1+), and (e) IHC-based unclassifiable (AR-, CD8-, FOXC1-, DCLK1-). The κ statistic indicated substantial agreement between the IHC-based classification and mRNA-based classification. Multivariate survival analysis suggested that our IHC-based classification was an independent prognostic factor for relapse-free survival. Transcriptomic data and pathological observations implied potential treatment strategies for different subtypes. The IHC-LAR subtype showed relative activation of HER2 pathway. The IHC-IM subtype tended to exhibit an immune-inflamed phenotype characterized by the infiltration of CD8+ T cells into tumor parenchyma. The IHC-BLIS subtype showed high expression of a VEGF signature. The IHC-MES subtype displayed activation of JAK/STAT3 signaling pathway. CONCLUSION: We developed an IHC-based approach to classify TNBCs into molecular subtypes. This IHC-based classification can provide additional information for prognostic evaluation. It allows for subgrouping of TNBC patients in clinical trials and evaluating the efficacy of targeted therapies within certain subtypes. IMPLICATIONS FOR PRACTICE: An immunohistochemistry (IHC)-based classification approach was developed for triple-negative breast cancer (TNBC), which exhibited substantial agreement with the mRNA expression-based classification. This IHC-based classification (a) allows for subgrouping of TNBC patients in large clinical trials and evaluating the efficacy of targeted therapies within certain subtypes, (b) will contribute to the practical application of subtype-specific treatment for patients with TNBC, and (c) can provide additional information beyond traditional prognostic factors in relapse prediction.

Clinical Observation of Zikun Decoction in Treatment of Syndrome of Yin Deficiency and Blood Dryness in Delayed Menorrhea Cycle Due to Decreasing Ovarian Reservation / 中国实验方剂学杂志

Objective:To observe the effect of Zikun decoction (ZKD) in treatment of Yin deficiency and blood dryness in delayed menorrhea due to decreasing ovarian reservation．Method:A total of 60 cases were randomly divided into two groups．The observation group was given ZKD orally，and the control group was treated with complex packing estradiol tablets/estradiol and dydrogestero for three months．The control group began to take medicine on the first day of menstruation for 28 days, while the observation group began to take medicine on the fifth day of menstruation, and stopped taking medicine on the first day of the next menstruation. Three menstrual cycles were followed up. The clinical syndrome scores, follicle-stimulating hormone (FSH), luteinizing hormone (LH), inhibin B (INHB), FSH/LH, Estradiol (E2), anti-müllerian hormone (AMH), and antral follicle count (AFC) scores of the patients before and after treatment were compared.Result:After treatment, there was no significant difference in clinical efficiency between observation group and control group, but the traditional Chinese medicine syndrome score of observation group was significantly reduced (P<0.05), observation group and control group had the same curative effect in improvement of FSH, FSH/LH, AMH and AFC (P<0.05), but ZKD was better than complex packing estradiol tablets/estradiol and dydrogestero in improvement of INHB.Conclusion:ZKD has a good treatment effect in the treatment of DOR. It can effectively improve patients' sex hormone levels, increase antral follicle count, improve the ovarian reserve function and promote menstruation.

A hesperetin derivative plays a role in immunoregulatory effect on human macrophages.

The immune system is an important physiological defense system. Its balance and stability are closely related to the body's health. Once the immune system loses its dynamic balance, the immune response will be blocked, which will lead to the occurrence of various diseases. Hesperetin is a kind of natural flavonoids extracted from citrus fruits of Rutaceae and it has many pharmacological activities. However, its water solubility and liposolubility are poor, and it is easy to be quickly metabolized in vivo, so it is difficult to maintain high blood drug concentration. Therefore, its derivative (HES) was found by structural modification. In this study, THP-1 cells were used as experimental model to investigate the immunomodulatory effect of HES in vitro. The results showed that HES participates in immune response by enhancing phagocytosis of macrophages to promote the release of NO, IL-6 and IL-1ß, and enhancing immunity by up-regulating the expression of Bcl-2 and Bcl-XL proteins. This study provides a theoretical and practical basis for the development of HES as an immunomodulator in the future.

Simultaneous multi­gene mutation screening using SNPscan in patients from ethnic minorities with nonsyndromic hearing­impairment in Northwest China.

The present study aimed to investigate the molecular etiology of nonsyndromic hearing impairment (HI) in hearing impaired populations of Hui, Tibetan, and Tu ethnicities in northwest China. A total of 283 unrelated subjects with HI who attended special education schools in northwest China were enrolled in the present study. Single-nucleotide polymorphisms (SNPs) in three common deafness­related genes, gap junction protein ß2 (GJB2), solute carrier family 26 member 4 (SLC26A4) and mitochondrially encoded 12S RNA (mtDNA12SrRNA), were detected using a SNPscan technique. GJB2 mutations were detected in 14.89% of Hui patients, 9.37% of Tibetan patients and 11.83% of Tu patients. The most prevalent GJB2 mutation in the Hui and Tu patients was c.235delC. In the Tibetan patients, the c.109G>A SNP exhibited the highest allele frequency. SLC26A4 mutations were detected in 10.64% of Hui patients, 6.25% of Tibetan patients, and 8.6% of Tu patients. The most common SLC26A4 mutation was c.919­2A>Gin the Hui, Tibetan, and Tu patients, and the second most common SLC26A4 mutations in these patients were c.1517T>G, c.1226G>A andc.2168A>G, respectively. The mutation rates ofmtDNA12SrRNA in the Hui, Tibetan, and Tu patients were 1.06, 5.21, and 5.38%, respectively. These findings demonstrate that the mutation spectra of these deafness­related genes are unique amongst these three ethnic groups. This information will be helpful in designing a protocol for genetic testing for deafness and for achieving accurate molecular diagnoses in northwest China.

Risk of congenital anomalies associated with antithyroid treatment during pregnancy: a meta-analysis.

To evaluate the association of either propylthiouracil or methimazole treatment for hyperthyroidism during pregnancy with congenital malformations, relevant studies were identified by searching Medline, PubMed, the Cochrane Library and EMBASE. We intended to include randomized controlled trials, but no such trials were identified. Thus, we included cohort studies and case-control studies in this meta-analysis. A total of 7 studies were included in the meta-analyses. The results revealed an increased risk of birth defects among the group of pregnant women with hyperthyroidism treated with methimazole compared with the control group (odds ratio 1.76, 95% confidence interval 1.47-2.10) or the non-exposed group (odds ratio 1.71, 95% confidence interval 1.39-2.10). A maternal shift between methimazole and propylthiouracil was associated with an increased odds ratio of birth defects (odds ratio 1.88, 95% confidence interval 1.27-2.77). An equal risk of birth defects was observed between the group of pregnant women with hyperthyroidism treated with propylthiouracil and the non-exposed group (odds ratio 1.18, 95% confidence interval 0.97-1.42). There was only a slight trend towards an increased risk of congenital malformations in infants whose mothers were treated with propylthiouracil compared with in infants whose mothers were healthy controls (odds ratio 1.29, 95% confidence interval 1.07-1.55). The children of women receiving methimazole treatment showed an increased risk of adverse fetal outcomes relative to those of mothers receiving propylthiouracil treatment. We found that propylthiouracil was a safer choice for treating pregnant women with hyperthyroidism according to the risk of birth defects but that a shift between methimazole and propylthiouracil failed to provide protection against birth defects.

Risk of congenital anomalies associated with antithyroid treatment during pregnancy: a meta-analysis

To evaluate the association of either propylthiouracil or methimazole treatment for hyperthyroidism during pregnancy with congenital malformations, relevant studies were identified by searching Medline, PubMed, the Cochrane Library and EMBASE. We intended to include randomized controlled trials, but no such trials were identified. Thus, we included cohort studies and case-control studies in this meta-analysis. A total of 7 studies were included in the meta-analyses. The results revealed an increased risk of birth defects among the group of pregnant women with hyperthyroidism treated with methimazole compared with the control group (odds ratio 1.76, 95% confidence interval 1.47-2.10) or the non-exposed group (odds ratio 1.71, 95% confidence interval 1.39-2.10). A maternal shift between methimazole and propylthiouracil was associated with an increased odds ratio of birth defects (odds ratio 1.88, 95% confidence interval 1.27-2.77). An equal risk of birth defects was observed between the group of pregnant women with hyperthyroidism treated with propylthiouracil and the non-exposed group (odds ratio 1.18, 95% confidence interval 0.97-1.42). There was only a slight trend towards an increased risk of congenital malformations in infants whose mothers were treated with propylthiouracil compared with in infants whose mothers were healthy controls (odds ratio 1.29, 95% confidence interval 1.07-1.55). The children of women receiving methimazole treatment showed an increased risk of adverse fetal outcomes relative to those of mothers receiving propylthiouracil treatment. We found that propylthiouracil was a safer choice for treating pregnant women with hyperthyroidism according to the risk of birth defects but that a shift between methimazole and propylthiouracil failed to provide protection against birth defects. .

Comparison of the pharmacokinetics of linezolid in burn and non-burn rabbits.

Linezolid is effective on many resistant organisms for the treatment of severe infections in burns. However, its pharmacokinetics was difficult to predict after major burns. The study aimed to describe the pharmacokinetic properties of linezolid administered intravenously at a dose of 10 mg/kg in severely burned rabbits in comparison to that in non-burns. Linezolid concentrations were quantitatively analyzed by high-performance liquid chromatography. The direct consequence of the physiological changes after burn injury was lower plasma linezolid concentrations. In addition, burn injury induced significantly altered pharmacokinetic parameters with higher inter-individual variability. The distribution volume and clearance rate were increased (2.88 vs. 1.92 L/kg, P > 0.05; 0.28 vs. 0.20 L/h/kg, P < 0.05), and the AUC0-∞ was significantly lower (37.99 vs. 51.47 mg/L h, P < 0.05). However, there were almost no changes in half-life and mean residence time. These results suggested that therapeutic drug monitoring and dosage individualization of linezolid in patients with severe burns were necessary.

Analysis of common deafness gene mutations in deaf people from unique ethnic groups in Gansu Province, China.

CONCLUSIONS: The GJB2 gene mutation characteristic of Dongxiang was the interaction result of ethnic background and geographical environment, and Yugur exhibited the typical founder effect. The SLC26A4 gene mutation characteristic of Dongxiang was related to caucasian backgrounds and selection of purpose exons, i.e. ethnic background and the penetrance of ethnic specificity caused the low mtDNA1555A>G mutation frequency in Dongxiang. OBJECTIVES: To determine the prevalence of GJB2 and SLC26A4 genes and mtDNA1555A>G mutations and analyze the ethnic specificity in the non-syndromic sensorineural hearing loss (NSHL) of unique ethnic groups in Gansu Province. METHODS: Peripheral blood samples were obtained from Dongxiang, Yugur, Bonan, and ethnic Han groups with moderately severe to profound NSHL in Gansu Province. Bidirectional sequencing (or enzyme digestion) was applied to identify the sequence variations. RESULTS: The pathogenic allele frequency of the three gene mutations was different. The frequency of the GJB2 gene among the Dongxiang, Yugur, Bonan, and ethnic Han groups was 9.03%, 12.5%, 5.88%, and 12.17%, respectively. No difference was found between the ethnic groups. The frequencies of the SLC26A4 genes were 3.23%, 8.33%, 0%, and 9.81%, respectively. The mutation frequency of mtDNA1555A>G was 0%, 0%, 0%, and 6.03%, respectively. No difference was found between the ethnic groups, except for the Dongxiang and ethnic Han groups, both in SLC26A4 gene and mtDNA1555A>G.

[Effect of Dingguier umbilical paste on rats with functional dyspepsia and mice with splenic asthenia].

OBJECTIVE: To observe the effect of Dingguier umbilical paste on rats with functional dyspepsia and mice with splenic asthenia, and investigate the related mechanism. METHOD: Functional dyspepsia models of rats were made by irregular food intake plus diluted hydrochloric acid. Successional treatments were offered for 14 days. The rats weights, contents of serum NO, AChE and MC were measured. The rats with splenic asthenia were made by rhubarb feed, and observed the affection of gastric emptying. RESULT: Compared with those in the model control group, the weight of rats in all dosages Dingguier umbilical paste groups increased obviously (P < 0.05), pepsin activity of rats in the dosage (1.34 g x kg(-1)) Dingguier umbilical paste groups was significantly higher and the contents of NO and quantities of MC in the dosage (2.67 g x kg(-1)) Dingguier umbilical paste groups decreased clearly (P < 0.05), and the contents of serum AChE in all dosages Dingguier umbilical paste groups rose apparently. The weight of mice with splenic asthenia increased obviously, accelerated gastric emptying, and improved the symptom. CONCLUSION: Dingguier umbilical paste has significant improvement of indigestion. The related mechanism may be to reduce the content of serum NO and the quantity of MC and enhance the content of serum AChE.

A randomized trial of Chinese medicine Lirukang Granule combined with psychological intervention for cyclomastopathy and menoxenia.

OBJECTIVE: To observe the influence of therapy with Chinese medicine Lirukang Granule (, LRKG) combined with psychological intervention on anxiety states and sex hormones in patients with cyclomastopathy and menoxenia. METHODS: A total of 470 subjects were randomly assigned to three groups by the net-central randomization system, the treatment group (161 patients, treated with LRKG and psychological intervention), the Chinese medicine group (157 patients, treated with LRKG), and the psychological intervention group (152 patients, treated with psychological intervention). The dose of LRKG was 12 g three times per day; psychological intervention included establishing relations, cognitive intervention and psychological persuasion, 30-40 min per session, once a week. The therapy duration for all groups was three months. The efficacy was compared and anxiety state/State-Trait Anxiety Invertory (STAI) scoring was measured before and after treatment. The serum estradiol (E2), progesterone (P), prolactin (PRL) and follicle stimulating hormone (FSH) levels of 60 patients selected randomly from each group during the luteal phase were measured before and after treatment, and a group of 20 healthy women were evaluated for comparison. A follow-up was arranged for one year after treatment. RESULTS: Thirty subjects were lost to follow-up. (1) Comparison of efficacy: the markedly effective rate and the total effective rate of the treatment group were 86.67% (131/150) and 98.00% (147/150), respectively; of the Chinese medicine group, 64.58% (93/144) and 90.27% (130/144), respectively; and of the psychological intervention group, 0% (0/146) and 3.42% (5/146), respectively. The markedly effective rate and the total effective rate in the treatment group were significantly higher than those in the Chinese medicine and psychological intervention groups (P < 0.05). (2) Comparison of STAI scoring: STAI scoring was decreased dramatically in the treatment group after treatment compared with that of the Chinese medicine group (P < 0.01), but there was no significant difference compared with the psychological intervention group. (3) Comparison of levels of sex hormones: E2, P, PRL and FSH of the three patient groups were disordered before treatment, and significantly different from healthy women (P < 0.01). After treatment, the levels of P and FSH of the treatment group were significantly increased (P < 0.01), E2 and PRL were significantly reduced, which were also significantly decreased compared with the psychological intervention groups (P < 0.01). (4) FOLLOW-UP: the markedly effective rate and the total effective rate of the treatment group remained higher than those of the other two groups after one year of treatment (P < 0.05). (5) Adverse reactions: no obvious adverse reactions were found among the three groups. CONCLUSIONS: Therapy with Chinese medicine combined with psychological intervention was effective for short-term and long-term treatment of cyclomastopathy and menoxenia. The mechanism might be related to the regulation of sex hormones.

[Experimental study on stabilization of sewage sludge by MOC].

Magnesium oxychloride cement (MOC) was used for the stability agent in the stabilization experiments of sewage sludge. It is found that MgCl2 in MOC is a kind of water-absorbent, water absorption of MgCl2 can be achieved at 1.55 mL/g (per 100 g sludge). Meanwhile, some water in sludge can be combined with MOC in the hydration reaction and sludge moisture content can be reduced efficaciously. The crystal structure of 3 phase and 5 phase, which occurred in the hydration process, makes the sludge compressive strength as high as 85.14 kg/cm2. The best ratio of MOC/sludge is 3/100, and MgO/MgCl2 is 3/1. Mg-Si-Al gel system is formed with Si2+, Al3+, Cu2+ in the sludge under alkaline condition, and it plays an important role in the stabilization of the heavy metals in sludge. Leaching experiments of sludge show that heavy metal concentrations of Cu, Zn, Cd, Cr, As in lixivium from sludge are lower than leachability standard.

Hemin modulates cytokine expressions in macrophage-derived foam cells via heme oxygenase-1 induction.

Lipid-laden foam cells were considered to be targets for therapeutic intervention in atherosclerosis. Several studies proposed new approaches to alter both lipid accumulation and inflammatory responses in macrophages. Finding anti-inflammatory signals during foam cell formation would provide new valid targets for anti-atherosclerotic treatment. The aim of the present study was to see whether oxidized low-density lipoprotein (ox-LDL) can active heme oxygenase (HO)-1 expression level in a human monocyte line, U937 cells, associated with the increase of cytokine secretion. We used hemin (HO-1 activator) and zinc protoporphyrin IX (ZnPP IX, HO-1 inhibitor) to determine the effect of HO-1 on the regulation of cytokine expressions. The results showed that hemin can significantly decrease pro-inflammatory cytokines interleukin (IL)-1beta and tumor necrosis factor (TNF)-alpha levels, while enhancing IL-10 production in a dose-dependent manner in U937 foam cells. ZnPP IX did not significantly affect cytokine levels in foam cells. Our present results suggested that HO-1 is an important anti-inflammatory therapeutic target through inhibiting pro-inflammatory cytokines and enhancing anti-inflammatory cytokines for the management of atherogenesis.

[Acceleration of burn wound healing with topical application of recombinant human epidermal growth factor ointments].

OBJECTIVE: To investigate the efficiency of recombinant human epidermal growth factor (rhEGF) on burn wound healing and to explore the effective density of the ointments. METHODS: A total of 120 cases of burn in superficial II degree and profound II degree were randomly divided into 2 groups. In the first group of 15 cases of superficial II degree, the wounds were treated by rhEGF ointments of different density, 0.5 microgram/g, 10 micrograms/g and 50 micrograms/g, to screen out the effective density. And in the other 105 cases of the second group, optimal density of the ointments based on the result of the first group were employed to treat the burn wound in superficial II degree and profound II degree, with the self-corresponding wounds of the same degree as control, to study the efficiency of rhEGF on wound healing, according to the wound healing time, and adverse reaction of the ointment. RESULTS: In the first group, the average healing time of superficial II wound treated by ointments of 10 micrograms/g and 50 micrograms/g significantly shortened when compared with that treated by ointments of 0.5 microgram/g(P < 0.01), but there was no obvious difference between the cases treated by ointments of 10 micrograms/g and 50 micrograms/g. In the second group, the healing time of superficial II wound treated by ointments of 10 micrograms/g was (8.39 +/- 2.25) days, (9.52 +/- 2.56) days in the control (P < 0.01); and healing time of profound II burn treated by ointments of 10 micrograms/g was (16.80 +/- 2.99) days, (18.27 +/- 3.17) days in the control (P < 0.01). And healing rates of burn wound at different periods were higher than those of the control. CONCLUSION: The above results indicate that rhEGF ointments can enhance burn wound healing significantly, and the ointment of 10 micrograms/g is a good choice for clinical application.