Exploring symptom clusters in Chinese patients with peritoneal dialysis: a network analysis.

BACKGROUND: In recent years, the research on symptom management in peritoneal dialysis (PD) patients has shifted from a single symptom to symptom clusters and network analysis. This study collected and evaluated unpleasant symptoms in PD patients and explored groups of symptoms that may affect PD patients with a view to higher symptom management. METHODS: The symptoms of PD patients were measured using the modified Dialysis Symptom Index. The symptom network and node characteristics were assessed by network analysis, and symptom clusters were explored by factor analysis. RESULTS: In this study of 602 PD patients (mean age 47.8 ± 16.8 years, 47.34% male), most had less than 2 years of dialysis experience. Five symptom clusters were obtained from factor analysis, which were body symptom cluster, gastrointestinal symptom cluster, mood symptom cluster, sexual disorder symptom cluster, and skin-sleep symptom cluster. Itching and decreased interest in sex may be sentinel symptoms, and being tired or lack of energy and feeling anxious are core symptoms in PD patients. CONCLUSIONS: This study emphasizes the importance of recognizing symptom clusters in PD patients for better symptom management. Five clusters were identified, with key symptoms including itching, decreased interest in sex, fatigue, and anxiety. Early intervention focused on these symptom clusters in PD patients holds promise for alleviating the burden of symptoms.

Transcriptome analysis of Mythimna separata: De novo assembly and detection of genes related to beta-cypermethrin resistance.

The oriental armyworm Mythimna separata (Walker) is a devastating pest of cereal crops mainly in Asia and Oceania and recently become resistant to beta-cypermethrin (beta-CP). However, molecular biological studies of its response to beta-CP are scarce, and related genomic information is not available. In this study, we sequenced and de novo assembled the transcriptome of beta-CP susceptible M. separata (MsS-whole, abbr. MsS-W). A total of 30,486 unigenes were obtained, with an N50 length of 506 bp. A number of 21,051 unigenes were matched to public databases, of which 10,107 were classified into 59 gene ontology subcategories, 5792 were assigned into 25 clusters of orthologous groups of proteins subgroups and 12,123 were assigned to 257 Kyoto Encyclopedia of Genes and Genomes pathways. A total of 729 simple sequence repeats were detected. In addition, a total of 323 cytochrome P450-associated sequences from nine lepidopterous species, of which 130 were from M. separata, were analyzed using the maximum likelihood method and Bayesian inference. Among the 130 cytochrome P450-associated sequences from M. separata, 60 were dropped into CYP3 clan, which is associated with metabolizing xenobiotics and plant natural compounds. Furthermore, the beta-CP susceptible (MsS-2) and resistant (MsR-2) M. separata population transcriptomes were sequenced. Certain critical genes involved in beta-CP detoxification were detected and verified by quantitative real-time polymerase chain reaction. Collectively, our results provided a basis for further studies of the molecular mechanism of insecticide resistance in M. separata.

Spiradiclisdetianensis (Rubiaceae, Ophiorrhizeae), a new species from southwestern Guangxi, China.

A new species of Rubiaceae, Spiradiclisdetianensis is described from a limestone karst area of southwestern China. This new species is morphologically similar to S.cordata and S.spathulata. All of them have rosetted habit and long peduncles, but it differs from the former by the cuneate leaf bases (vs. basally cordate) and much longer corolla tubes (1.8-2.2 cm long vs. ca. 5 mm long), and from the latter mainly by its tubular-funnel shaped corolla (vs. slenderly salver shaped), 4.5-6.8 (vs. 1.5-2) mm in diam, inside throat and corolla densely puberulent (vs. glabrous except a ring of long hairs at the middle). It also resembles to S.tubiflora, but differs clearly by its subrosulate habit (vs. procumbent to creeping), longer leaf blades (7.0-10.5 cm vs. 0.5-2.5 cm) and longer corolla tubes (18-22 mm vs. 14-16 mm). At same time, color photos, illustrations, detailed descriptions and conservation status of the new species are provided.

Oropharyngeal administration of colostrum for preventing necrotizing enterocolitis and late-onset sepsis in preterm infants with gestational age ≤ 32 weeks: a pilot single-center randomized controlled trial.

BACKGROUND: Oropharyngeal administration of colostrum (OAC) may provide immunoprotective and anti-inflammatory effects that potentially reduce the incidence of necrotizing enterocolitis (NEC) and late-onset sepsis and improve short-term outcomes. Our objective was to evaluate the role of OAC in the early prevention of NEC and late-onset sepsis in preterm infants with gestational age (GA) ≤ 32 weeks. METHODS: A pilot, single-center, 1:1 parallel randomized controlled trial was conducted in a 40-bed tertiary neonatal intensive care unit (NICU) in China from 1 January 2019 to 30 September 2020. Preterm infants were randomly divided into two groups with GA ≤ 32 weeks. The OAC group included preterm infants who received 0.4 ml of maternal colostrum via the oropharyngeal route every 3 h for 10 days beginning within the first 48 h after birth, and the control group included preterm infants who received normal saline instead. Data from the two groups were collected and compared. RESULTS: A total of 127 infants in the OAC group and 125 infants in the control group were enrolled. The incidence of NEC (Bell stage 2 or 3) and late-onset sepsis were lower in the OAC group [2.36% vs. 10.40%, relative risk (RR) 0.23 (95% confidence interval (CI) 0.07, 0.78), adjusted RR 0.23 (95% CI 0.06, 0.84); 4.72% vs. 13.60%, RR 0.35 (95% CI 0.14, 0.85), adjusted RR 0.36 (95% CI 0.14, 0.95)]. In addition, the incidence of proven sepsis and intraventricular hemorrhage (IVH) (stage 3 or 4) were lower in the OAC group [2.36% vs. 8.80%, RR 0.27 (95% CI 0.08, 0.94); 1.57% vs. 7.20%, RR 0.22 (95% CI 0.05, 0.99)], and the time to achieve full enteral feeding was shorter (23.13 ± 9.45 days vs. 28.50 ± 14.80 days). No adverse reactions were observed in either group. CONCLUSIONS: Oropharyngeal administration of colostrum is a safe and simple NICU procedure that may yield a potential effect in decreasing the incidences of NEC, late-onset sepsis, and severe IVH and shorten the time to achieve full enteral feeding in preterm infants with GA ≤ 32 weeks. TRIAL REGISTRATION: Chinese Clinical Trial Registry, ChiCTR1900023697 , Registered 8 June 2019, retrospectively registered.

Leptomischus hiepii, a new species of Rubiaceae from Vietnam.

Leptomischus hiepii, a new species of the tribe Argostemmateae from Son La province, northwestern Vietnam, is described and illustrated. Morphologically it allies to L. wallichii, L. erianthus and L. funingensis by sharing a similar habit, large stipules and similar corolla shape, but it differs by its anisophyllous leaves, 1-flowered inflorescences, homostylous flowers and tubular-campanulate corollas.

Downregulation of Notch1 inhibits the invasion and metastasis of human gastric cancer cells SGC7901 and MKN74 in vitro through PTEN activation and dephosphorylation of Akt and FAK.

Migration and invasion are both vital causes of mortality in patients with gastric cancer. Therefore, the inhibition of these tumour cell processes is of great importance in gastric cancer therapy. Activation of Notch has been reported in many cancers. The critical role of Notch and its regulation in tumourigenesis has been noted. Although the studies on Notch in the field of cancer have been performed extensively, the role of Notch1 signalling in gastric cancer requires further study. Inactivation of PTEN has been observed in the development of many malignant tumors, and loss of PTEN function has been implicated in tumorigenic processes. Notch acts as an upstream signalling pathway that regulates PTEN activities. However, the effect of Notch on invasion and metastasis in gastric cancer and the regulation of PTEN during this process remain poorly understood. In the present study, small interfering RNA (siRNA) was used to knock down Notch1 expression in gastric cancer cell lines SGC7901 and MKN74. The mRNA and protein expression of Notch1, PTEN, Akt and FAK were measured upon depletion of Notch1. phospho­PTEN, phospho­Akt and phospho­FAK expression were measured using western blot analysis. Migration and invasion assays were also used after Notch1 depletion. Our results showed that the knockdown of Notch1 leads to the inhibition of cell invasion and metastasis of human gastric cancer cells SCG7901 and MKN74 in vitro. Compared to control and mock groups, PTEN activities were significantly promoted following depletion of Notch1, and the expression of Phospho­Akt and Phospho­FAK were downregulated. Taken together, our findings suggest that Notch1 could be used as a therapeutic target to inhibit cell invasion and migration in gastric cancer.

Alanine and arginine rich domain containing protein, Aard, is directly regulated by androgen receptor in mouse Sertoli cells.

Alanine and arginine rich domain containing protein (Aard) is specifically expressed in Sertoli cells (SCs) of mouse testis and the expression increases in an age­dependent manner. A number of previous studies have indicated that androgen and androgen receptor (AR) signaling pathways are particularly important for spermatogenesis in mouse SCs, however, the association between Aard and AR remain to be elucidated. The present study identified Aard as a gene that is directly regulated by AR in mouse SCs, which is important in spermatogenesis. The expression of AARD was significantly downregulated in the testes of Sertoli cell­selective AR knockout mice compared with wild­type mice as analyzed by western blotting and immunofluorescence analyses. Quantitative polymerase chain reaction and western blotting indicated that AARD was predominantly expressed in adult mouse testis and its expression was increased in an age-dependent manner. In addition, AARD expression was upregulated by testosterone in primary SCs in vitro, which was confirmed by bioinformatics analysis and a dual­luciferase reporter assay. Finally, chromatin immunoprecipitation and electrophoretic mobility shift assays indicated that the ligand­bound AR activated Aard transcription via directly binding to the androgen­responsive element of the Aard promoter. To the best of our knowledge, the present study is the first to document that Aard is directly regulated by AR in mouse Sertoli cells.

Downregulation of Notch1 inhibits the invasion of human hepatocellular carcinoma HepG2 and MHCC97H cells through the regulation of PTEN and FAK.

Tumor invasion and metastasis are the main causes of mortality in patients with hepatocellular carcinoma (HCC). Thus, the effective inhibition of these tumorigenic processes is critical in order for HCC therapy to be effective. Previous studies have demonstrated that Notch1 is associated with metastasis in several human malignancies. However, the exact molecular mechanisms underlying the Notch1-mediated induction of the invasion of HCC cells remain poorly understood. In the present study, we demonstrate that, compared to the normal liver cell line, L02, Notch1 is highly expressed in the human HCC cell lines, HepG2 and MHCC97H. Using small interfering RNA (siRNA), we knocked down the expression of Notch1 in the cell lines. Notch1 expression in the HCC cell lines was also measured following transfection with siRNA using RT-PCR and western blot analysis. In addition, a migration and invasion assay was performed to determine the effects of Notch1 knockdown on cell migration and invasion. Our results demonstrated that the downregulation of Notch1 by small interfering RNA (siRNA) significantly inhibited the migration and invasion of both HCC cell lines. Additionally, we demonstrated that the knockdown of Notch1 in both HCC cell lines increased both the total expression of phosphatase and tensin homolog (PTEN) and its phosphorylated form. By contrast, focal adhesion kinase (FAK) and phospho-FAK expression was decreased following Notch1 depletion. Taken together, our data suggest that targeting Notch1 may be a useful therapeutic approach to inhibiting the metastasis of HCC cells.

Notch signaling: a novel regulating differentiation mechanism of human umbilical cord blood-derived mesenchymal stem cells into insulin-producing cells in vitro.

BACKGROUND: Human umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs) could be induced to differentiate into insulin producing cells (IPCs) in vitro, which have good application potential in the cell replacement treatment of type-1 diabetes. However, the mechanisms regulating this differentiation have remained largely unknown. Notch signaling is critical in cell differentiation. This study investigated whether Notch signaling could regulate the IPCs differentiation of human UCB-MSCs. METHODS: Using an interfering Notch signaling protocol in vitro, we studied the role of Notch signaling in differentiation of human UCB-MSCs into IPCs. In a control group the induction took place without interfering Notch signaling. RESULTS: Human UCB-MSCs expressed the genes of Notch receptors (Notch 1 and Notch 2) and ligands (Jagged 1 and Deltalike 1). Human UCB-MSCs with over-expressing Notch signaling in differentiation resulted in the down-regulation of insulin gene level, proinsulin protein expression, and insulin-positive cells percentage compared with the control group. These results showed that over-expressing Notch signaling inhibited IPCs differentiation. Conversely, when Notch signaling was attenuated by receptor inhibitor, the induced cells increased on average by 3.06-fold (n = 4, P < 0.001) in insulin gene level, 2.60-fold (n = 3, P < 0.02) in proinsulin protein expression, and 1.62-fold (n = 6, P < 0.001) in the rate of IPCs compared with the control group. Notch signaling inhibition significantly promoted IPCs differentiation with about 40% of human UCB-MSCs that converted to IPCs, but these IPCs were not responsive to glucose challenge very well both in vitro and in vivo. Hence, further research has to be carried out in the future. CONCLUSIONS: Notch signaling may be an important mechanism regulating IPCs differentiation of human UCB-MSCs in vitro and Notch signaling inhibition may be an efficient way to increase the number of IPCs, which may resolve the shortage of islet of cell replacement treatment of type-1 diabetes.

A secretory function of human insulin-producing cells in vivo.

BACKGROUND: Mesenchymal stem cells derived from human umbilical cord blood (UCB-MSCs) have good research and application prospects in the treatment of diabetes. We once induced UCB-MSCs to differentiate into insulin-producing cells (IPCs) in vitro, but we did not know the functions of these cells in vivo. The aim of this study was to assess the functional effects of IPCs on insulin secretion and their role in the treatment of diabetes in vivo. METHODS: UCB-MSCs were induced to IPCs by an inducing protocol with extracellular matrix gel. BALB/C nude mice were made hyperglycemic by intraperitoneal injection of streptozotocin. The diabetic mice were transplanted with 1X10(7) IPCs under the renal capsule or with phosphate-buffered saline as a control. After transplantation, the grafts were analyzed by immunocytochemistry for the expression of human insulin; the serum human insulin levels were measured; and blood glucose and body weight status were monitored. RESULTS: Immunofluorescence showed that numerous IPCs under the kidney capsule were insulin-positive. On day 14 after transplantation, the serum human insulin level of the treatment group (n=9) averaged 0.44+/-0.12 mU/L, which was higher than that of the control group (n=9) that did not express insulin (t=10.842, P<0.05). The diabetic mice remained hyperglycemic and kept losing body weight after IPC transplantation, and there was no significant difference in the control group. CONCLUSION: IPCs differentiated from UCB-MSCs generate human insulin in diabetic mice, but more research is needed to make further use of them to regulate hyperglycemia and body weight in vivo.

Extracellular matrix gel is necessary for in vitro cultivation of insulin producing cells from human umbilical cord blood derived mesenchymal stem cells.

BACKGROUND: Pancreatic islet cell transplantation is an effective approach to treat type 1 diabetes. However, this therapy is not widely used because of the severe shortage of transplantable donor islets. This study investigated whether mesenchymal stem cells (MSCs) derived from human umbilical cord blood (UCB) could be transdifferentiated into insulin producing cells in vitro and the role of extracellular matrix (ECM) gel in this procedure. METHODS: Human UCB samples were collected and MSCs were isolated. MSCs specific marker proteins were analyzed by a flow cytometer. The capacities of osteoblast and adipocyte to differentiate were tested. Differentiation into islet like cell was induced by a 15-day protocol with or without ECM gel. Pancreatic characteristics were evaluated with immunofluorescence, reverse transcription polymerase chain reaction (RT-PCR) and flow cytometry. Insulin content and release in response to glucose stimulation were detected with chemiluminescent immunoassay system. RESULTS: Sixteen MSCs were isolated from 42 term human UCB units (38%). Human UCB-MSCs expressed MSCs specific markers and could be induced in vitro into osteoblast and adipocyte. Islet like cell clusters appeared about 9 days after pancreatic differentiation in the inducing system with ECM gel. The insulin positive cells accounted for (25.2 +/- 3.4)% of the induced cells. The induced cells expressed islet related genes and hormones, but were not very responsive to glucose challenge. When MSCs were induced without ECM gel, clusters formation and secretion of functional islet proteins could not be observed. CONCLUSIONS: Human UCB-MSCs can differentiate into islet like cells in vitro and ECM gel plays an important role in pancreatic endocrine cell maturation and formation of three dimensional structures.

In vitro cultivation of islet-like cell clusters from human umbilical cord blood-derived mesenchymal stem cells.

A major obstacle to successful islet transplantation for both type 1 and 2 diabetes is an inadequate supply of insulin-producing tissue. In vitro transdifferentiation of human umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs) into insulin-producing cells could provide an abundant source of cells for this procedure. For this study, we isolated and characterized human UCB-MSCs and induced them in vitro to differentiate into islet-like cell clusters using a 15-day protocol based on a combination of high-glucose, retinoic acid, nicotinamide, epidermal growth factor, and exendin-4. These clusters appeared about 9 days after pancreatic differentiation; expressed pancreatic beta-cell markers, including insulin, glucagon, Glut-2, PDX1, Pax4, and Ngn3; and could synthesize and secrete functional islet proteins at the end of the inducing protocol. The insulin-positive cells accounted for (25.2-3.36)% of whole induced cells. Although insulin secretion of those insulin-producing cells did not respond to glucose challenge very well, human UCB-MSCs have the ability to differentiate into islet-like cells in vitro and may be a potential new source for islet transplantation.

[Preliminary report of surgical treatment of vernal keratoconjunctivitis with giant papillae].

OBJECTIVE: To evaluate the efficacy and safety of excision and cryotherapy combine with amniotic membrane transplantation (AMT) for the treatment of vernal keratoconjunctivitis (VKC) with giant papillae (GP). METHODS: Eight patients (16 eyes) with VKC, characterized by GP on the upper tarsal conjunctiva, refractory to medical treatment, underwent excision and cryotherapy associated with AMT. The follow up period ranged 3-22 months. RESULTS: Corneal shield ulcers and superficial punctuate keratitis healed during the first week after the surgery and did not recur. Fourteen eyes were symptom free and without GP one month after surgery. No ectropion, trichiasis and other complications were noted. The blood vessels of upper tarsal conjunctiva could not be seen clearly, a small amount of conjunctival scar was observed. Recurrence of GP was observed in 2 eyes, with reduced area and lessened irritation symptom as compared with preoperative status. Among them, one eye was treated by cyclosporine eyedrops with improvement. The other eye did not improve, and underwent secondary surgery (a cotton patch soaked in fluorouracil was applied on the upper eyelid after excision and cryotherapy). Nine months after the treatment, the patient had no symptoms and GP did not recur. CONCLUSIONS: Excision and cryotherapy combine with AMT may be an effective and safe method for the treatment of refractory VKC with GP. But the candidates for surgical treatment should be chosen carefully.

[Experimental research of the expression of telomerase in corneal limbal epithelial cells].

OBJECTIVE: To explore the possibility of using telomerase as a marker of corneal limbal stem cells. METHODS: Corneal limbal tissues and central corneal epithelial tissues from 8 rabbits were examined for telomerase activity qualitatively by telomere repeat amplification protocol (TRAP) and quantitatively by detecting the light value with bioluminescent technique. 5-Fu (20 mg) was injected subconjunctivally in 4 rabbits and the other 4 rabbits were not injected. RESULTS: Telomerase activity was positive in all corneal limbal tissues and negative in all central corneal tissues. Telomerase activity of corneal limbal cells (light value 165,575) was significantly higher than that of central corneal epithelial cells (light value 34,912) by bioluminescent technique (P = 0.001). 5-Fu injection group showed higher telomerase activity than that of the group without injection (light value 145,754) (P = 0.02). CONCLUSIONS: Positive telomerase activity is detected in the corneal limbal tissues. It suggests that there are cells with high proliferative ability in the corneal limbus. Telomerase activity may be used as a marker of corneal limbal stem cells.

[Phototherapeutic keratectomy combined with photorefractive keratectomy for the treatment of myopia with corneal scars].

OBJECTIVE: To evaluate the effects of phototherapeutic keratectomy (PTK) combined with photorefractive keratectomy (PRK) in the treatment of myopia with corneal scars. METHODS: Corneal epithelium was removed with laser plus scraping (d=7.0 mm). Corneal scars were removed with PTK (d=7.0 mm) and followed by PRK for myopia (d=6.0 mm). Healon was used to make corneal surface smoother during operation. RESULTS: 79 eyes of 54 cases of myopia with corneal scars were followed up at least for one year. Mean refractive degree was (-6.73 +/- 4.17) D and corrected vision was 0.63 before operation. After operation,corneal scars in 50 eyes (63.3%) were removed in operative zone and corneal haze was less than I degree. The vision acuity in 69 eyes (89.9%) was equal to or better than preoperative best correction. Mean postoperative vision was 0.74. Corneal surface was smoother and astigmatism was reduced after surgery. CONCLUSION: PTK combined with PRK is a safe and effective treatment for myopia with corneal scars.

[Interferon alpha-2b inhibits rabbit corneal haze induced by PRK].

OBJECTIVES: To evaluate the effect of interferon alpha-2b on the treatment of corneal haze after PRK. METHODS: 36 rabbits were divided into three groups (A.B.C). The refractive degree of ablation was -4.00 D, -8.00 D, -10.00 D in each group respectively. The treatment given in group A was 1 x 10(5) IU/ml interferon alpha-2b, in group B 0.1% dexamethasone and placebo eyedrops for group C after PRK 2 months. Corneal haze, histopathologic response, intraocular pressure (IOP) were investigated. RESULTS: Less corneal haze was observed in group A and B. Postoperative IOP was higher than that of preoperation in group B. Keratocytes and fibrotic response in corneal stroma were active up to 2 months in group C. CONCLUSIONS: Interferon alpha-2b was effective on inhibition of corneal haze after PRK.