[Multidisciplinary regenerative treatment and mechanisms for rescuing a severe calciphylaxis patient with human amnion-derived mesenchymal stem cells].

Calciphylaxis is a rare disease with severe pain and high-mortality due to cutaneous ischemic necrosis and infection that currently lacks proved effective therapies. The occurrence of calciphylaxis in end stage kidney disease (ESKD) patients is known as calcific uremic arteriolopathy (CUA), which is characterized histologically by dermal microvessel calcification, intimal fibroplasia and microthrombosis. Here we innovatively treated a severe CUA patient with human amnion-derived mesenchymal stem cells (hAMSCs). A 34-year-old uremic woman was presented with progressive, painful malodorous ulcers in buttocks and mummified lower limbs. Skin pathological features supported the diagnosis of calciphylaxis. The patient was refractory to conventional multidisciplinary symptomatic therapies. With the approval of our hospital ethics committee, she was treated with hAMSCs including intravenous and local intramuscular injection, and external application of hAMSC culture supernatant to the wound area. During 15-month follow-up, the patient had regeneration of skin and soft tissues, with improved blood biochemical, inflammatory, mineral and bone metabolic indices and immunoregulation effects. After 15-month hAMSC treatment, the score of pain visual analog scale (VAS) decreased from 10 to 0, Bates-Jensen wound assessment tool (BWAT) score decreased from 65 to 13, and wound-quality of life (Wound-QoL) questionnaire score decreased from 68 to 0. We propose that hAMSC treatment is promising for CUA patients. The therapy is potentially involved in the multiple beneficial effects of inhibiting vascular calcification, stimulating angiogenesis and myogenesis, modulating adverse inflammatory and immunologic responses, promoting re-epithelialization and restoring skin integrity.

[Clinical features of 141 fatal cases of coronavirus disease in Jinyintan Hospital in Wuhan, China].

Objective: To describe the epidemiological characteristics and clinical features of patients with fatal coronavirus disease (COVID-19), in order to provide evidence for clinical diagnosis and treatment. Methods: In this retrospective study, we analyzed data on 141 fatal cases of confirmed COVID-19 that occurred among patients in Jinyintan Hospital in Wuhan, China, from January 20 to March 6, 2020. We analyzed their epidemiological characteristics, clinical and radiological features, laboratory results, and treatment. Results: Of the 141 patients (49 females, 92 males), the median age was 77 years (range: 24-92 years). The most likely source of exposure included the Huanan seafood market (n=3, 2%), family members (n=6, 4%), and hospital-acquired infection (n=8, 6%). The remaining 116 patients (72%) had no known source of exposure. Of the patients, 101 (72%) had chronic diseases. The most common comorbidities were hypertension, diabetes and coronary heart disease. The most common clinical manifestations were fever (n=121, 85%), dry cough (n=77, 54%), shortness of breath (n=23, 16%), and chest pain (n=15, 10%). Less common clinical manifestations included fatigue (n=7, 4%), headache (n=3, 2%), disorders of consciousness (n=2, 1%), diarrhea (n=2, 1%) and lumbago (n=1, 0.7%). In terms of laboratory tests, the absolute value of lymphocytes in most patients was reduced (n=132, 94%), but C-reactive protein (n=141, 100%), procalcitonin(n=121, 89%), serum amyloid (n=140, 99%) were significantly increased. The most common findings on imaging of the lungs were bilateral multiple mottling and ground-glass opacity (n=101, 72%), mainly in the lower lobes (n=15, 10%), with lesions being more common on the right. Other imaging findings included diffuse consolidation (n=4, 3%), ground-glass opacity and consolidation (n=20, 14%), and pneumothorax (n=1, 0.7%). All patients were treated with antibiotics and antiviral drugs. Other treatments included immunoglobulin (n=49, 35%), corticosteroids (n=45, 32%), continuous renal replacement therapy (n=24, 17%), and extracorporeal membrane oxygenation (n=12, 9%). All patients were treated with oxygen therapy. The mode of administration included invasive mechanical ventilation (n=61, 43%), noninvasive mechanical ventilation (n=65, 46%), and nasal catheter oxygen inhalation (n=15, 11%). The direct causes of death were acute respiratory distress syndrome (n=90, 64%), multiple organ failure (n=24, 17%), sudden cardiac arrest (n=11, 8%), viral myocarditis (n=8, 5%), acute myocardial infarction (n=4, 3%), cerebrovascular accident (n=3, 2%), and acute gastrointestinal bleeding (n=1, 0.7%). Conclusions: Risk factors for death due to COVID-19 included older age, male sex, and the presence of comorbidities. The most common direct causes of death were acute respiratory distress syndrome, multiple organ failure, sudden cardiac arrest, and viral myocarditis.

[Enteral nutrition support reduces toxicity of chemotherapy in patients with advanced or metastatic esophageal cancer].

OBJECTIVE: To assess the impact of enteral nutrition support on response and toxicity of the first-line chemotherapy in those patients with advanced or metastatic esophageal cancer. METHODS: We collected the clinical data of 118 patients with unresectable advanced or metastatic esophageal cancer who received the first-line chemotherapy in our center from July 2014 to December 2016. All these 118 esophageal cancer patients were then divided into two groups: the nutrition group (received enteral nutrition support in addition to chemotherapy) and the control group (received chemotherapy only). Differences were analyzed before and after chemotherapy in each of the nutritional indicators including Karnofsky performance status (KPS), weight, body mass index (BMI), hemoglobin (Hb), number of lymphocytes (Lymph), total protein (TP), albumin (Alb), triglycerides (TG), total cholesterol (TC) in both groups. And differences of the efficacy and toxicities of the first-line chemotherapy between the two groups were also analyzed. RESULTS: (1) Weight, BMI and Hb were all significantly decreased after chemotherapy in the control group (P<0.001), while there was no significant change of weight and BMI in the nutrition group, just with Hb decrease only. However, there was no significant change of all the other nutrition indicators after chemotherapy in both groups. (2) Compared with the control group, the nutrition group had significantly lower incidence of grade 3 to 4 hematologic toxicities after chemotherapy (15.4% vs. 42.1%, P=0.004). In addition, the incidence of grade 3 to 4 nonhematologic toxicities after chemotherapy was also lower in the nutrition group but without statistical significance (0 vs. 9.2%, P=0.123). Logistic regression model was then used for multivariate analysis to identify the factors that affected the toxicity of chemotherapy in these patients, and the results showed that nutrition therapy was an independent influencing factor of grade 3 or higher hematological toxicity after chemotherapy in the patients with esophageal cancer (P=0.008, RR=6.048, 95%CI: 1.589-23.027). (3) The response rate of chemotherapy between the control group and the nutrition group had not significant difference. CONCLUSION: Enteral nutrition support in addition to chemotherapy could improve nutrition status and reduce toxicity of chemotherapy in advanced or metastatic esophageal cancer patients.

[CD20-positive T cell lymphoma: clinicopathological features of five cases].

Objective: To investigate the clinicopathological characteristics of the T cell lymphomas with CD20 expression, and to better understand this rare entity. Methods: Two-hundred cases of T-cell lymphoma diagnosed in the Department of Pathology of the Affiliated Hospital of Qingdao University from November 2016 to February 2020 were examined, and 5 cases of CD20-positive T-cell lymphomas were identified and included. Combined with clinical data and review of the literature, the clinicopathological characteristics of the disease were analyzed. Results: The five patients were all male, and had an average age of 56 years (range, 47 to 64 years). There were 2 cases of monomorphic epitheliotropic intestinal T-cell lymphoma, 2 cases of mycosis fungoides (1 case was plaque stage and the other was tumor stage) and 1 case of indolent T-cell lymphoproliferative disorder of the gastrointestinal tract. Immunohistochemistry showed that all 5 cases expressed multiple T cell markers (CD3/CD4/CD5/CD7/CD8) and only one of B cell markers (CD20). Three of the 5 cases were negative for CD20 at the first diagnosis, while CD20 was diffusely positive on the second biopsy from the recurrence or progression of the disease, without expression of CD79a or PAX5. Epstein-Barr encoding region (EBER) in situ hybridization was negative in all 5 cases. T-cell receptor gene analysis showed monoclonal rearrangement of ß or/and δ chains;Ig rearrangements were all polyclonal. None of the five patients were treated with rituximab, and 4 patients survived with disease and 1 patient survived without disease at the end of follow-up. Among them, the patient with mycosis fungoides at the cancerous stage has progressed rapidly and had poor quality of life. Conclusions: CD20-positive T-cell lymphoma is extremely rare. Its prognosis is closely related to the type of T-cell lymphoma, clinical stage and initial therapeutic effect. However, the expression of CD20 indicates the recurrence or progression of the disease, and the prognosis is relatively poor. When CD3 expression is absent in T-cell lymphoma, it is easy to be misdiagnosed as B-cell lymphoma. The combination of multiple immunohistochemical antibodies and molecular detection can improve the accuracy of diagnosis.

Identification and interaction analysis of key genes and microRNAs in atopic dermatitis by bioinformatics analysis.

BACKGROUND: Atopic dermatitis (AD) is a skin disease that carries a major health burden, but the exact mechanism of the disease is not yet known. AIM: To identify the key genes and micro (mi)RNAs in AD, and to explore their potential molecular mechanisms. METHODS: From the Gene Expression Omnibus (GEO) database, we downloaded microarray data for GSE32924 (mRNA profile) and GSE31408 (miRNA profile), which were analysed using GEO2R. Functional and pathway enrichment analyses were performed using the DAVID database, and the protein-protein interaction network was constructed with Cytoscape software. In addition, targets of differentially expressed miRNAs (DEMs) were predicted by the online resource miRDB. RESULTS: In total, 328 differentially expressed genes (DEGs) were identified, including 121 upregulated and 207 downregulated genes. Gene Ontology analyses showed that upregulated genes were significantly enriched in immune responses, while downregulated genes were mainly involved in epidermis development. In addition, we identified three DEMs, all of which were downregulated. Hsa-let-7a-5p may target CCR7, and hsa-miR-26a-5p probably targets HAS3. CONCLUSIONS: We identified lists of DEGs and DEMs in AD. Bioinformatics and interaction analysis may provide new clues for further studies of AD.

[Preliminary results of femtosecond laser-assisted Descemet stripping endothelial keratoplasty for bullous keratopathy].

OBJECTIVE: To evaluate the preliminary results of femtosecond laser-assisted Descemet stripping endothelial keratoplasty (FS-DSEK) for the treatment of bullous keratopathy. METHODS: In the nonrandomized, prospective, consecutive case series, 7 eyes with bullous keratopathy induced by cataract surgery received FS-DSEK surgery between September and December 2013. Standard Descemet stripping endothelial keratoplasty procedure was performed. The Descemet membrane and abnormal endothelial layer were stripped from the central recipient posterior surface in a diameter of 7.75 mm. The graft was fixed to the edge of recipient cornea with one single stitch. The donor lenticule was produced by Intralase 150 femtosecond laser with a diameter of 7.75 mm. The adherence of donor lenticule to the recipient posterior stroma and postoperative donor lenticule dislocation were monitored in the early stage after surgery. Best spectacle-corrected visual acuity, refraction, endothelial cell density and thickness of grafts were measured preoperatively, and at 1 week, 1, 3, 6 and 12 months after FS-DSEK. RESULTS: Two eyes had graft dislocation, with a gap between the graft and recipent posterior stroma at 2 days after surgery. The grafts were repositioned after bubbles were added into the anterior chamber. All grafts were well adherent to the recipent posterior stromal surface at 1 week postoperatively. One graft failed at 3 months postoperatively, and penetrating keratoplasty was performed. At 12 months, the best spectacle-corrected visual acuity was 0.04 to 0.7, the mean endothelial cell density was (1 698.0±251.8) cells/mm(2), and the mean thickness of grafts was (124.2 ± 11.4) µm. CONCLUSION: Femtosecond laser-assisted Descemet stripping endothelial keratoplasty was effective in treating bullous keratopathy. Stitching the graft to the recipent cornea can prevent postoperative graft dislocation effectively.

Life and death of human vascular endothelial cells by stimulation with free fatty acids through death receptors.

Diabetic individuals may have elevated levels of serum free fatty acids and may exhibit injury to the vascular endothelial cells. This study was undertaken to determine the relationship between various free fatty acids (FFAs) and vascular endothelial cell injury and the molecular mechanisms linking FFA-induced vascular endothelial cells injury or protection. We observed the survival of HUVECs exposed to different FFAs, and our results revealed that the effects of various FFAs on the cell survival of HUVECs were significantly different. Palmitic acid (PA) markedly decreased the HUVEC survival rate in a time- and dose-dependent manner, but arachidonic acid (AA) significantly increased the cell survival rate and could partially prevent cellular apoptosis induced by PA. Interestingly, PA and AA could activate the same target receptor, TNF-R1. PA induced the apoptosis of HUVECs by initiating the death pathway (TNF-R1/TRADD/caspases 8 pathway), whereas AA enhanced cell survival to protect vascular endothelial cells by activating the survival pathway (TNF-R1/RIP/NF-κB 50/NF-κB 65).

LncRNA TUG1 influences osteoblast proliferation and differentiation through the Wnt/ß-catenin signaling pathway.

OBJECTIVE: Long non-coding ribonucleic acids (lncRNAs) play a vital role in bone development, but the function of lncRNA taurine up-regulated gene 1 (TUG1) in osteoblast proliferation and differentiation is still unknown. MATERIALS AND METHODS: The expression of TUG1 and the messenger RNA (mRNA) expressions of the Wnt/ß-catenin signaling pathway markers [Runt-related transcription factor 2 (Runx2), Frizzled-2, axis inhibition protein 2 (Axin 2) and ß-catenin] at 0 d, 1 d, 7 d and 14 d after in vitro culture of osteoblasts were detected, respectively, by Reverse Transcription-Polymerase Chain Reaction (RT-PCR). The effects of TUG1 on the Wnt/ß-catenin signaling pathway markers and osteoblast proliferation and differentiation were studied through the silencing of TUG1 by short hairpin TUG1 (shTUG1). Furthermore, the effects of the Wnt/ß-catenin signal on osteoblast proliferation and differentiation was verified by Wnt/ß-catenin signal inhibitors. RESULTS: With the continuous differentiation of osteoblasts, the level of TUG1 was significantly increased. The mRNA levels of the Wnt/ß-catenin signaling pathway markers (Runx2, Frizzled-2, Axin 2 and ß-catenin) also showed the same increasing trend. ShTUG1 notably reduced the activity of alkaline phosphatases (ALPs), the levels of osteocalcin and osteopontin and osteoblast proliferation activity. In addition, the silencing of TUG1 by shTUG1 resulted in significant reductions in the proteins of the Wnt/ß-catenin signaling pathway markers (Runx2, Frizzled-2, Axin 2 and ß-catenin), and Wnt/ß-catenin inhibitors markedly reduced osteoblast proliferation and differentiation activity. CONCLUSIONS: LncRNA TUG1 inhibition can suppress the Wnt/ß-catenin signaling pathway and reduce osteoblast proliferation and differentiation.

[Application of the five-level pediatric emergency triage system: a single center study].

Objective: To assess the effectiveness in optimizing resources and shortening critical children's waiting time in pediatric emergency department (PED) with five-level pediatric emergency triage system (PETS). Methods: This retrospective study was conducted in the First Affiliated Hospital of Xiamen University after PETS was applied. The data of patients who visited the pediatric emergency department from January 2015 to December 2017 were collected and analyzed, including age, sex, diseases, visiting time, triage rate and destination. Results: A total of 375 985 patients were included, among whom males were 225 308 (59.9%) and females were 150 677 (40.1%), all younger than 14 years of age. The number of critical cases (level Ⅰ, level Ⅱ and level Ⅲ) was increased from 4 719 (3.7%) in 2015, 12 209 (10.2%) in 2016 to 16 188 (12.7%) in 2017. The number of non-critical patients (level Ⅴ) decreased year by year, as from 98 213 (76.8%) in 2015 to 75 210 (62.6%) in 2016 and 78 857 (61.7%) in 2017. The patients who classified as level Ⅰ or levelⅡaccording to the PETS were seen immediately by physician (n=1855, 0.5%). Overall, 119 738 patients (98.3%) who were classified as level Ⅲ or level Ⅳ could be seen by physician in a timely manner according to triage guidelines, while 2 112 patients (1.7%) could not. The mean waiting time was 9.09 min in level Ⅲ, 17.7 min in level Ⅳ, and 55.76 min in level Ⅴ patients, respectively. The critical cases admitted to the intensive care units were 175 (36.2%) in 2015, 350 (62.8%) in 2016 and 374 (66.2%) in 2017. The etiologies were respiratory diseases (73.3%), gastrointestinal diseases (15.8%) and infectious diseases (3.1%). Conclusion: The application of PETS could optimize emergency resources and shorten the waiting time of critically ill children.

Two novel Co(II) complexes with two different Schiff bases: inhibiting growth of human skin cancer cells.

Using two flexible Schiff bases, H2L1 and H2L2, two new cobalt II (Co(II))-coordination compounds, namely, Py3CoL1 (1) and Py3CoL2 (2) (Py=pyridine, L1=3,5-ClC6H2(O)C=NC6H3(O)-4-NO2, L2=3,5-BrC6H2(O)C=NC6H3(O)-4-NO2) have been synthesized under solvothermal conditions. Single crystal X-ray structural analysis revealed that compounds 1 and 2 are both six-coordinate in a distorted octahedral geometry, and the 1D chain structure was formed by the π…π and C-H…O interactions or C-H…Cl interaction. The in vitro antitumor activities of 1, 2 and their corresponding organic ligands Py, L1, and L2 were studied and evaluated, in which three human skin cancer cell lines (A-431, HT-144 and SK-MEL-30) were used in the screening tests.

The role and clinical applications of bioactive lysolipids in ovarian cancer.

OBJECTIVE: To review the current understanding of the role of bioactive lysolipids in ovarian cancer and their potential clinical applications. METHODS: A MEDLINE search and our own work, including some unpublished work, are the major sources of the review. The MEDLINE search terms used included lysophosphatidic acid, lysophophatidylcholine (LPC), lysophosphatidylinositol (LPI), sphingosine-1-phosphate, and sphingosylphosphorylcholine (SPC). RESULTS: Elevated lysolipid levels were detected in plasma and ascites samples from patients with ovarian cancer compared with samples from healthy controls or patients with nonmalignant diseases. These lysolipids regulate growth adhesion, production of angiogenic factors, and chemotherapeutic drug resistance in ovarian cancer cells. Ovarian cancer cells were likely to be at least one of the sources for elevated lysolipid levels in the blood and ascites of patients with ovarian cancer. CONCLUSIONS: Bioactive lysolipid levels might be sensitive markers for detecting gynecologic cancers, particularly ovarian cancer. The prognostic value of lysolipids in ascites is worth further investigation. Bioactive lysolipid molecules can affect both the proliferative and metastatic potentials of ovarian cancer cells; therefore, regulation of the production or degradation of these lipids and interception of the interaction between these lipids and their receptors could provide novel and useful preventative or therapeutic measures.

[A study of potential risk of HBsAg negative with RPHA].

An experimental study of potential risk of hepatitis B virus infection was carried out on 1,074 donors who were HBsAg negative by RPHA. Those so-called "qualified donors" were reexamined by ELISA and SPRIA for HBsAg, HBeAg, anti-HBc and anti-HBc-IgM, and 89 of them examined for HBV-DNA by spot hybridization. The results showed that their positive rates of HBsAg, anti-HBc-IgM, HBeAg and HBV-DNA were 5.96%, 3.35%, 2.14% and 65.17% respectively, with a total positive rate of 9.68%. The present so-called "qualified donors" are undoubtedly playing the role of source of infection to those who need blood transfusion.

Herb-drug interactions: methods to identify potential influence of genetic variations in genes encoding drug metabolizing enzymes and drug transporters.

Herbal supplements are often used concomitantly with conventional medications resulting in considerable potential for herb-drug interactions. These interactions, which are generally through interfering with pharmacokinetic and/or pharmacodynamic pathways, may result in beneficial effects or more often adverse reactions such as toxicity or treatment failure and may be influenced by multiple environmental and/or genetic factors. The pharmacogenetic approach may help to identify some interactions which may be more pronounced or only occur in specific groups of subjects although the complex nature of the herbal medicines may limit the discovery of such an interaction. Preclinical studies such as gene expression profiling in rodent liver may help to define metabolic pathways influenced by herbal medicines and facilitate more accurate targeting of human in vivo studies. This review discusses the mechanisms of herb-drugs interaction and the potential influence of genetic variation on herb-drug interactions based on available clinical evidence.

Two novel Co(II) complexes with two different Schiff bases: inhibiting growth of human skin cancer cells

Using two flexible Schiff bases, H2L1 and H2L2, two new cobalt II (Co(II))-coordination compounds, namely, Py3CoL1 (1) and Py3CoL2 (2) (Py=pyridine, L1=3,5-ClC6H2(O)C=NC6H3(O)-4-NO2, L2=3,5-BrC6H2(O)C=NC6H3(O)-4-NO2) have been synthesized under solvothermal conditions. Single crystal X-ray structural analysis revealed that compounds 1 and 2 are both six-coordinate in a distorted octahedral geometry, and the 1D chain structure was formed by the π…π and C-H…O interactions or C-H…Cl interaction. The in vitro antitumor activities of 1, 2 and their corresponding organic ligands Py, L1, and L2 were studied and evaluated, in which three human skin cancer cell lines (A-431, HT-144 and SK-MEL-30) were used in the screening tests.

[Seroepidemiological study of HBV infection in Hunan area].

A seroepidemiological study of HBV infection was carried out in a "normal population" of 3,089 in Hunan area by means of SPRIA and ELISA in 1983-1986. The positivity rates of HBsAg, anti-HBs and anti-HBc were 17.87%, 31.76% and 73.03% respectively. The overall rate of HBV infection was 80.77%, All the positive rates of HBsAg, anti-HBs and anti-HBc peak at the age of 10-15 years, then, it remains on a similar level. This results indicate that Hunan is a high epidemic area of HBV and the age of persons at peak infection rate is very young. About half of HBsAg positive persons is HBeAg positive and a part of persons who are both HBsAg and HBeAg positive has the lesion of liver function. Thus these persons could be the reservoirs of HBV infection and it was necessary to treat and deal with them seriously.

Lysophospholipids increase interleukin-8 expression in ovarian cancer cells.

OBJECTIVES: We have previously described that bioactive lysophospholipids-lysophosphatidic acid (LPA), sphingosine 1-phosphate (S1P), and sphingosylphosphorylcholine (SPC)-are present in ascitic fluids from patients with ovarian cancer. To understand the role of these lipids in ovarian cancer, we investigated the effects of these lipids on interleukin-8 (IL-8) production in ovarian cancer cells. IL-8 is a proinflammatory and proangiogenic factor, which is potentially involved in ovarian cancer development. METHODS: The Clontech PCR-Select cDNA subtraction method (Clontech Laboratories, Inc., Palo Alto, CA) was used to identify genes potentially regulated by LPA in HEY and OCC1 ovarian cancer cell lines. Northern blot analysis was used to confirm and examine IL-8 mRNA regulation by lysolipids. Enzyme-linked immunosorbent assay (ELISA) was used for detecting secreted IL-8. RESULTS: We describe here that LPA, S1P, and SPC increased mRNA levels (2- to 7-fold) and protein secretion (2- to 12-fold) of IL-8 from ovarian cancer cells (HEY, OCC1, and SKOV3) in vitro. These regulations were both dose- and time-dependent. All three lipids increased the stability IL-8 mRNA in HEY cells. In contrast to malignant ovarian cancer cells, immortalized human ovarian epithelial cells did not respond to any of these lipids to increase the secretion of IL-8, although these cells secreted similar basal levels of IL-8 (310 pg/ml/10,000 cells). Two breast cancer cell lines (MCF7 and T47D) secreted lower basal levels of IL-8 (48-80 pg/ml/10,000 cells), compared with ovarian cancer cells (200-500 pg/ml/10,000 cells). MCF7 cells responded to LPA, but not S1P and SPC, by increasing the secretion of IL-8. T47D and MCF10A, an immortalized breast cell line, did not respond to LPA, S1P, or SPC to increase IL-8 secretion. CONCLUSIONS: LPA, S1P, and SPC regulate the mRNA and protein levels of the proinflammatory and proangiogenic factor IL-8 in ovarian cancer cells. The pathological significance of these regulations in ovarian cancer remains to be further investigated.