Impaired wound healing is a common complication of diabetes mellitus and a major morbidity that leads to pain and severely diminished quality of life. Diabetic wounds are commonly associated with defective immune cell responses or abnormality of extracellular matrix. Various types of electrical stimulation interventions have been used to promote tissue healing. However, it is unclear whether high-voltage pulsed current stimulation (HVPCS) enhances diabetic wound healing. In this study, the effects of HVPCS on wound healing were investigated in diabetic rats. Three groups of rats (10 per group) were used: non-diabetic control, diabetic control, and diabetic rats that were administered HVPCS for 40 minutes daily for 1 week. Rats from control groups were administered sham interventions. Dorsal incision wounds were generated in all animals, and wound-healing rate was determined during one-week intervention. After interventions, we measured the relative expression levels of collagen type I (collagen-I), α-smooth muscle actin (α-SMA), and transforming growth factor-β1 (TGF-β1) mRNAs in the wounded skin. Wound closure was delayed in diabetic control rats compared to the non-diabetic control rats, and the diabetic control rats showed the reduced expression levels of collagen-I, α-SMA and TGF-β1 mRNAs. Importantly, compared to diabetic control rats, rats with HVPCS showed accelerated wound closure and healing (p < 0.01) and restored expression levels of collagen-I (p = 0.02), α-SMA (p = 0.04), and TGF-β1 (p = 0.01) mRNAs. In conclusion, HVPCS may be beneficial for enhancing the healing of diabetic wounds by restoring the expression levels of TGF-β1, collagen-I, and α-SMA.
REM sleep behavior disorder (RBD) is one type of parasomnia characterized by nocturnal complex motor activity associated with dream mentation. Growing evidence has indicated that RBD is a preclinical stage of neurodegenerative diseases. Therefore screening RBD patient is becoming important. The RBD Questionnaire-Hong Kong (RBDQ-HK) is an effective questionnaire to screen RBD patients. However, it is hard to distinguish RBD with the questionnaire from severe OSAS patients, who could mimic some symptoms of RBD patients. Therefore, we made RBDQ-Beijing by adding two screening questions for OSAS into original RBDQ-HK, including habitual loud snoring and witnessed apnea during sleep. To validate and compare these two questionnaires, 224 subjects were enrolled and screened with these questionnaires, and consequently analyzed with video-polysomnography. Receiver-operator characteristics curve analysis was conducted to attain the best cut-off values of the RBDQ-HK and RBDQ-Beijing. For the RBDQ-HK, the sensitivity was 97.1% and the specificity was 83.2%. More than half of misclassified RBD patients were proved to be severe OSAS patients. For the RBDQ-Beijing, the sensitivity was 95.8% and specificity was 94.3%, indicating that our questionnaire is able to distinguish RBD from severe OSAS patients. In conclusion, RBDQ-Beijing is of help to improve the specificity in RBD screening without excluding the patients with RBD combined OSAS. Therefore the RBDQ-Beijing may be a better screening and preliminary diagnostic tool for RBD than the RBDQ-HK. Moreover, the RBDQ-Beijing would be important for early diagnosis of neurodegenerative diseases and for prevention of injuries to the patient or the patient’s bed partner.
Keratoconus (KC) is a non-inflammatory eye disease characterized by progressive corneal thinning and asymmetrical conical protrusion of the cornea. Fuchs endothelial corneal dystrophy (FECD) is a degenerative, slowly progressive disease of the corneal endothelium that is characterized by alteration in corneal endothelial cell morphology and progressive loss of these cells. They are unrelated eye diseases that may ultimately lead to vision loss. Their pathogenesis is largely unknown, although impaired apoptosis has been suggested to be responsible for both diseases. Therefore, we studied the frequency of the c.–671A>G polymorphism of the apoptosis-related FAS gene and the c.–844T>C polymorphism of the FAS ligand (FASLG) gene in patients with FECD (221 individuals) or KC (264) and controls (300). Each polymorphism is located within the putative cis-acting element of the respective promoter. Risk of KC or FECD was estimated with unconditional multiple logistic regression with adjustment for various factors, including age, sex, allergies, and family history. The T/T genotype and the T allele of the c.–844T>C polymorphism were associated with increased occurrence of KC, while the C allele was associated with decreased KC occurrence. The G allele of the c.–671A>G polymorphism was associated with increased occurrence of FECD, while the A allele was associated with decreased FECD occurrence. The C/C-A/A combined genotype was associated with reduced risk of FECD, whereas the T/T-G/A combined genotype increased risk of KC. In conclusion, variability in the expression of the FAS and FASLG genes may be involved in the pathogenesis of KC and FECD.
Circulating tumor cells (CTCs) have been implicated in cancer prognosis and follow up. Detection of CTCs was considered significant in cancer evaluation. However, due to the heterogeneity and rareness of CTCs, detecting them with a single maker is usually challenged with low specificity and sensitivity. Previous studies concerning CTCs detection in lung cancer mainly focused on non-small cell lung carcinoma. Currently, there is no report yet describing the CTC detection with multiple markers in lung adenocarcinoma. In this study, by employing quantitative real-time PCR, we identified four candidate genes (mRNA) that were significantly elevated in peripheral blood mononuclear cells and biopsy tissue samples from patients with lung adenocarcinoma: cytokeratin 7 (CK7), Ca2+-activated chloride channel-2 (CLCA2), hyaluronan-mediated motility receptor (HMMR), and human telomerase catalytic subunit (hTERT). Then, the four markers were used for CTC detection; namely, positive detection was defined if at least one of the four markers was elevated. The positive CTC detection rate was 74.0% in patients with lung adenocarcinoma while 2.2% for healthy controls, 6.3% for benign lung disease, and 48.0% for non-adenocarcinoma non-small cell lung carcinoma. Furthermore, in a three-year follow-up study, patients with an increase in the detection markers of CTCs (CK7, CLCA2, HMMR or hTERT) on day 90 after first detection had shorter survival time compared to those with a decrease. These results demonstrate that the combination of the four markers with specificity and sensitivity is of great value in lung adenocarcinoma prognosis and follow up.
Maternal undernutrition can affect offspring’s physical status and various health parameters that might be transmittable across several generations. Many studies have focused on undernutrition throughout pregnancy, whereas maternal undernutrition prior to pregnancy is not sufficiently studied. The objective of our study was to explore the effects of food restriction prior to and during pregnancy on body weight and longevity of the second generation offspring. Adult female Wistar rats (“F0” generation) were 50% food restricted for one month prior to pregnancy (pre-pregnancy) or during pre-pregnancy and pregnancy. The third group was fed normally (control). The first generation offspring were normally fed until the 6th month of age to produce the second generation offspring; namely, the first-generation female rats were mated with male breeders from outside the experiment. The second generation offspring thus obtained were observed until natural death (up to 36 months). Compared to the controls, the second-generation male offspring whose “grandmothers (F0 females)” undernourished only during pre-pregnancy were significantly heavier from the 8th month of age, whereas no significant weight difference was found in the male offspring whose “grandmothers” were food-restricted during pre-pregnancy and pregnancy. Shorter lifespan was observed in the second-generation male offspring of “grandmothers” that were food-restricted either during pre-pregnancy or during pre-pregnancy and pregnancy. By contrast, no differences in body weight and lifespan were observed in all second-generation female offspring. In conclusion, maternal caloric restriction prior to pregnancy increases the body weight and shortens the longevity of the second-generation male offspring, indicating the sex-dependent transgenerational effect of maternal caloric restriction.
Systemic lupus erythematosus (SLE) is a systemic autoimmune disease that affects women of childbearing years. Pregnancy with SLE is associated with an increase risk of adverse maternal and fetal outcomes. Recently, tacrolimus has been used for steroid-resistant SLE. However, because of limited information regarding the use of immunosuppressants during pregnancy, many SLE patients give up pregnancy. We report two SLE patients receiving tacrolimus who hoped to become pregnant. Patient 1 was a 31-year-old woman diagnosed with SLE 6 years earlier and treated with tacrolimus for 3 years because her symptoms were not controlled with other immunosuppressants. Patient 2 was a 28-year-old woman diagnosed with SLE 13 years earlier and treated with tacrolimus for 3 years because her symptoms were not controlled with prednisolone alone. The medical ethics board in our hospital approved the use of tacrolimus during pregnancy and lactation, and informed consent was obtained from the patients. Both patients were well controlled during pregnancy with prednisolone (Patient 1: 12 mg/day and Patient 2: 10 mg/day) and tacrolimus therapy (3 mg/day). They had healthy newborns and continued breastfeeding with tacrolimus therapy. The blood concentrations of tacrolimus 12 hours after taking tacrolimus was 3.0 ng/ml in Patient 1 and 2.9 ng/ml in Patient 2, and their newborns’ blood concentrations of tacrolimus 1 hour after breastfeeding were 0.2 ng/ml and 0.5 ng/ml, respectively. Both newborns are healthy for at least 3 years after birth. This is the first report on the safety of tacrolimus for pregnancy and lactation in patients with SLE.
Immune cell Toll-like receptors (TLRs) recognize conserved microbial components, leading to immune and inflammatory responses. However, TLRs are also expressed in cancer cells, including melanoma cells, which express TLR2-4. TLR4 ligands have received attention as immunotherapies; therefore, we assessed the expression of TLR4 in human melanoma specimens (29 primary lesions and 28 metastatic lesions) representing different types of melanoma. A high percentage (≥ 90%) of melanoma lesions expressed TLR4, as judged by immunohistochemistry. Next, the role of TLR4 in cell proliferation and migration was assessed using the TLR4-positive (TLR4+) melanoma cell lines 501mel and 888mel, and TLR4-negative (TLR4‒) 928mel melanoma cells. Lipopolysaccharide (LPS), a TLR4 agonist, increased the proliferation of TLR4+ melanoma cells but not of TLR4‒ 928mel cells. The proliferation-inducing effect of LPS in 888mel cells was abolished by blockade of TLR4 signaling via treatment with short interfering RNA (siRNA) targeting TLR4 or myeloid differentiation primary response gene 88 (MyD88), a molecule downstream of TLR4. However, knockdown of TLR4 or MyD88 expression did not affect the LPS-induced proliferation of 501mel cells, suggesting that residual TLR4 signaling is sufficient to maintain cell proliferation. By contrast, LPS increased the migration of TLR4+ melanoma cells, and this effect was substantially inhibited by TLR4 or MyD88 knockdown. Furthermore, TLR4 knockdown decreased cell migration even in the absence of LPS, suggesting the presence of an endogenous TLR4 ligand(s) in melanoma cells. TLR4 signaling may contribute to melanoma progression, and caution should be exercised when using TLR4 ligands as adjuvant therapy for cancer.
Targeted anticancer therapies have been developed to interfere with specific target molecules including those of downstream pathways required for tumor growth and progression. Mammalian target of rapamycin (mTOR) has been considered as one of the target molecules of cancer growth, and its inhibitors have been reported to exert an anticancer effect in various malignant tumors. The pulmonary disorder is one of the major side effects of anticancer drugs including mTOR inhibitor (mTORi), and the diagnosis of lung injury induced by medication is difficult because of non-specific nature of the radiological findings. In this study, we present the detailed autopsy findings of a patient who developed diffuse alveolar damage (DAD) following mTORi treatment for metastatic renal cell carcinoma. We also studied 19 cases of DAD derived from other diseases and 9 cases with non-pathological lung. Of interest, pneumocytes of the patients with DAD, who received other anticancer drugs or contacted bacteria, demonstrated significantly lower mTOR activities than pneumocytes of those with non-pathological lung tissue, as judged by the immunohistochemical analysis. In contrast, both pneumocytes and T cells in DAD tissues of the patient treated with mTORi showed higher mTOR activities than those of patients with DAD of other causes, suggesting that the enhanced mTOR signaling may be involved in the development of DAD after mTORi treatment. This unexpected finding needs to be confirmed in other patients treated with mTORi. In conclusion, the attenuated mTOR signaling in pneumocytes may contribute to the pathogenesis of DAD in patients without mTORi treatment.
Every year, 14 million cases of obstetric hemorrhage occur worldwide, causing 127,000 maternal deaths. About 75% of postpartum hemorrhage cases are due to atonic uterus, which is loss of uterine muscular tone or strength for contraction of the uterus after delivery. The prediction of atonic uterus is therefore important for the prevention of postpartum maternal death. However, prediction of occurrence of atonic uterus is difficult before delivery, because the precise pathophysiological mechanism to trigger this condition remains unclear. Here, we present a case of severe postpartum hemorrhage due to atonic uterus. A 35-year-old woman gave a birth by vaginal delivery to a healthy boy. However, due to intractable massive hemorrhage after the removal of the retained placenta, we performed supravaginal hysterectomy as the best option for survival. Pathological examination showed that implantation site intermediate trophoblasts (ISITs) formed unusually large clumps in the decidua, diagnosed as exaggerated placental site (EPS). EPS is thought to be a condition consisting of an excessive number of ISITs. ISITs are differentiated from a trophoblast lineage in the process of placenta formation. ISITs anchor the placenta to the maternal tissue and are considered to maintain pregnancy, but the postpartum role of these cells remains unclear. Excessive infiltration of ISITs, namely EPS, may cause postpartum atonic uterus. In this article, we also reviewed the literatures on EPS. The present case and other reported cases indicate that EPS causes mass formation in the uterus, continuous uterine bleeding, and massive hemorrhage, resulting in hysterectomy.
Biliary tract cancers include cancers of the gallbladder and extrahepatic bile ducts, and its prognosis is poor. The anterior gradient 2 (AGR2) is a protein disulfide isomerase and is highly expressed in various human cancers, such as breast, prostate and pancreas cancers. AGR2 is expressed in normal cholangiocytes and its expression is maintained during biliary carcinogenesis. However, the clinical significance of AGR2 expression in biliary tract cancer has not yet been assessed. Thus, we examined the expression of AGR2 protein in biliary tract tumors using immunohistochemistry and its association with various clinicopathologic parameters. This study included 100 patients who underwent surgery for biliary tract cancers: 46 men and 54 women with a mean and median age of 64.2 and 65.0 years, respectively. AGR2 expression was detected in ductal epithelial cells of the normal biliary tract and in 95% of biliary tract cancer tissues. While the AGR2 expression was not associated with cancer location, patient age, patient sex, degree of regional lymph node metastasis (N-status), or residual status, the AGR2 expression level was decreased with increased tumor size (T-status, p = 0.006) and progression of tumor stage (p = 0.009). Moreover, well-differentiated cancers tended to show higher AGR2 expression than poorly differentiated cancers (p = 0.068); in fact, AGR2 expression was not associated with patient survival (Kaplan-Meier analysis, p = 0.415). Thus, AGR2 is of limited value as a prognostic marker for biliary tract cancer. In conclusion, the expression of AGR2 is decreased with the progression of biliary tract cancer.