[Analysis of AVPR2 variant in a neonate with congenital nephrogenic diabetes insipidus].

OBJECTIVE: To detect potential variant in a male neonate affected with congenital nephrogenic diabetes insipidus (CNDI). METHODS: Clinical data of the patient was collected. Genomic DNA was extracted from peripheral blood samples from the child and his parents. The whole coding regions of the arginine vasopressin V2 receptor (AVPR2) gene were amplified by PCR and subjected to Sanger sequencing. RESULTS: The patient presented recurrent fever and polyuria after birth. Multiple blood gas analyses indicated hypernatremia. Ultrasound showed bilateral hydronephrosis and hydroureter. The patient was partially responsive to hydrochlorothiazide. DNA analysis identified a hemizygous frameshift variant c.890-899delACCCGGAGGC in exon 2 of the AVPR2 gene in the proband. His mother was heterozygous for the same variant. CONCLUSION: The c.890-899delACCCGGAGGC variant of the AVPR2 gene probably underlies the CNDI in the child. Above discovery has enriched to spectrum of CNDI associated variants.

H-ferritin-nanocaged doxorubicin nanoparticles specifically target and kill tumors with a single-dose injection.

An ideal nanocarrier for efficient drug delivery must be able to target specific cells and carry high doses of therapeutic drugs and should also exhibit optimized physicochemical properties and biocompatibility. However, it is a tremendous challenge to engineer all of the above characteristics into a single carrier particle. Here, we show that natural H-ferritin (HFn) nanocages can carry high doses of doxorubicin (Dox) for tumor-specific targeting and killing without any targeting ligand functionalization or property modulation. Dox-loaded HFn (HFn-Dox) specifically bound and subsequently internalized into tumor cells via interaction with overexpressed transferrin receptor 1 and released Dox in the lysosomes. In vivo in the mouse, HFn-Dox exhibited more than 10-fold higher intratumoral drug concentration than free Dox and significantly inhibited tumor growth after a single-dose injection. Importantly, HFn-Dox displayed an excellent safety profile that significantly reduced healthy organ drug exposure and improved the maximum tolerated dose by fourfold compared with free Dox. Moreover, because the HFn nanocarrier has well-defined morphology and does not need any ligand modification or property modulation it can be easily produced with high purity and yield, which are requirements for drugs used in clinical trials. Thus, these unique properties make the HFn nanocage an ideal vehicle for efficient anticancer drug delivery.

A 10-year retrospective study of neonatal lupus erythematous in China.

BACKGROUND: Neonatal lupus erythematosus (NLE) is not a common disease. The death rate of complete congenital heart block (CCHB), which is the most severe clinical manifestation, is as high as 20% to 30%, so early recognition of infants at risk is important. OBJECTIVES: To investigate the clinical features and long-term prognosis of NLE. METHODS: Twenty-five cases with NLE were reviewed. The clinical manifestations of patients and their mothers were summarized and analyzed. Autoantibodies were detected, and long-term follow-up was carried out. RESULTS: There were 25 patients (male:female ratio of 11:14). CCHB was detected in only 3 of the 25 patients (12%). Cutaneous neonatal lupus erythematosus (CNLE) was seen in 22 of the 25 patients (88%). Eight babies were treated with intravenous immunoglobulin (IVIG), five of whom had a prolonged PR interval that reverted to normal sinus rhythm. During the follow-up of the patients, we found only two patients with CCHB without a pacemaker, who both exhibited growth delay. One patient with CCHB without a pacemaker died. CONCLUSIONS: Children with NLE have an excellent outcome when only skin lesions are present. Even the hepatic, hematological and neurological abnormalities are transient, with generally good outcomes. IVIG might have some effectiveness due to enhanced anti-inflammatory activity to treat early diseases that may be reversible (e.g. prolonged PR interval). The long-term prognosis for patients with NLE is still under investigation, and some infants with NLE may progress to other autoimmune diseases later in childhood.

Risk factors and clinical analysis for invasive fungal infection in neonatal intensive care unit patients.

OBJECTIVE: In this study, we seek to determine independent risk factors of invasive fungal infection (IFI) in neonatal infants. STUDY DESIGN: The medical charts of 5135 neonatal intensive care unit admissions in the past 7 years between January 2004 and December 2010 were reviewed and 45 neonates were found with IFI. Two controls, matched by gestational age, birth weight category, admission date, ward, hospital stay, and admission age, were selected for each case. RESULTS: Candida parapsilosis was the leading causative pathogen of IFI and was isolated in 33.3% of the patients. The mortality rate of the case group was 8.9% versus 1.1% in controls (p < 0.05). Multivariable logistic regression modeling defined intubation > 6 days (71.1%), use of peripherally inserted central venous catheter (68.8%), use of third-generation cephalosporin (53.3%), any prior abdominal surgeries (20.0%), and neutropenia during first week of life < 1.5 · 109/L (20.0%) as exposures significantly associated with case status. CONCLUSIONS: The predominant factors identified with IFI were third-generation cephalosporin use, peripherally inserted central venous catheter use, intubation > 6 days, any prior abdominal surgery, and neutropenia during first week of life < 1.5 · 109/L.

TiO2 supported single Ag atoms nanozyme for elimination of SARS-CoV2.

The outbreak of SARS-coronavirus 2 (SARS-CoV2) has become a global health emergency. Although enormous efforts have been made, there is still no effective treatment against the new virus. Herein, a TiO2 supported single-atom nanozyme containing atomically dispersed Ag atoms (Ag-TiO2 SAN) is designed to serve as a highly efficient antiviral nanomaterial. Compared with traditional nano-TiO2 and Ag, Ag-TiO2 SAN exhibits higher adsorption (99.65%) of SARS-CoV2 pseudovirus. This adsorption ability is due to the interaction between SAN and receptor binding domain (RBD) of spike 1 protein of SARS-CoV2. Theoretical calculation and experimental evidences indicate that the Ag atoms of SAN strongly bind to cysteine and asparagine, which are the most abundant amino acids on the surface of spike 1 RBD. After binding to the virus, the SAN/virus complex is typically phagocytosed by macrophages and colocalized with lysosomes. Interestingly, Ag-TiO2 SAN possesses high peroxidase-like activity responsible for reactive oxygen species production under acid conditions. The highly acidic microenvironment of lysosomes could favor oxygen reduction reaction process to eliminate the virus. With hACE2 transgenic mice, Ag-TiO2 SAN showed efficient anti-SARS-CoV2 pseudovirus activity. In conclusion, Ag-TiO2 SAN is a promising nanomaterial to achieve effective antiviral effects for SARS-CoV2.

[Association between clinical manifestations of infants with human cytomegalovirus infection and glycoprotein B genotype].

OBJECTIVE: To investigate the association between the clinical manifestations of infants with human cytomegalovirus (HCMV) infection and glycoprotein B (gB) genotype. METHODS: Urine samples were obtained from 107 symptomatic infants with HCMV infection confirmed by fluorescence quantitative PCR, 70 male and 37 female, aged 5 d-8 months, and 25 asymptomatic infants with HCMV infection, 16 male and 9 female, aged 21 d-7 months. A fragment of glycoprotein B gene was amplified by nested PCR (nPCR). HCMV gB genotyping was carried out by restriction fragment length polymorphism (RFLP), and some of the amplified DNA fragments were verified by DNA sequencing. RESULTS: Of the HCMV specimens from 107 infants, 53 were typed as gB group I, 20 as gB II, 18 as gB III, 7 as gB I and gB II, 5 as gB I and gB III, and 4 as gB II and gB III, and gB IV was not found. The HCMV gB genotype from 53 infants with hepatic function damage showed that 36 were classified as gB I, 5 as gB II, 7 as gB III, 3 as gB I and gB II, and 2 as gB I and gB III. The gB I genotype was more common among the infants with hepatic function damage (P < 0.05). The distribution of gB genotype in the asymptomatic infants was as follow: gB I, 10/25; gB II, 6/25; gB III, 8/25; and gB I and gB II, 1/25. The homology of PCR products of HCMV gB in 24 strains amplified compared with the sequences of prototype strains in GenBank was from 97.06% to 99.64%. CONCLUSION: RFLP analysis of HCMV gB genotype is definite and reliable. The gB I genotype is more common among the infants with hepatic function damage.

[Effects of phytoestrogens on gap junctional intercellular communication].

OBJECTIVE: To observe effects of phytoestrogens quercetin (QC), Genistein (GEN), coumestrol (COM), and enterolactone (ENL) on gap junctional intercellular communication (GJIC) in HaCaT cells. METHODS: HaCaT cells were exposed to QC, GEN, COM, and ENL at 0.1, 1.0, 10.0 and 100.0 micromol/L for 24 hours. The effects of phytoestrogens on GJIC were determined by fluorescence redistribution after photobleaching (FRAP) technique of using a laser scanning confocal microscope (LSCM). RESULTS: QC did not affect the GJIC at 0.1-10.0 micromol/L, whereas, GEN, COM, and ENL exhibited inhibition on the GJIC in some extent at 0.1-10.0 micromol/L without showing significant cytotoxicity. The ratio of fluorescence recovery were between 31.77% to 37.06%, which were significantly decreased compared the vehicle control (44.74%). CONCLUSION: The phytoestrogens GEN, COM, and ENL, but not QC, could inhibit the GJIC function in HaCaT cells at concentrations could be reached in human serum in some instance, indicating they could, under certain conditions, be cancer promoters. Therefore, it should be prudent to use these chemicals as pharmaceuticals or dietary supplements.

[Relationship among serum cortisol and insulin and blood glucose in the asphyxiated neonates].

OBJECTIVE: To explore the dynamic change of serum cortisol and insulin levels, and their relation with blood glucose concentration in asphyxiated neonates. METHODS: The levels of serum cortisol and insulin at d1,d3 and d7 of birth were measured by radioimmunoassay and the concentration of blood glucose was measured with glucose oxidase method in 43 asphyxiated neonates. RESULTS: The levels of serum cortisol at d 1, d 3 and d 7 of birth were gradually decreased (P<0.01). At d1, the incidence of hyperinsulism (>20 mIU/L) was 60.5%. The level of serum insulin reached normal level (

A novel RING finger E3 ligase RNF186 regulate ER stress-mediated apoptosis through interaction with BNip1.

Disturbances in the normal functions of the endoplasmic reticulum (ER) can lead to the accumulation of unfolded proteins and disturbance of Ca(2+) regulation within the lumen of ER, and arouse a series of complicated response termed unfolded protein response (UPR), which is aimed initially at reestablishing homeostasis and normal physiology but can ultimately trigger cell death if the UPR fails to compensate for damage. Here we show that ER locating human RING finger E3 ligase RNF186 participates in the process of ER stress-mediated apoptosis. Overexpression of RNF186 stimulates upregulation of ER sensor proteins and rapid transmission of ER Ca(2+) in Hela cells, while RNF186 knockdown exhibits a moderate degree of resistance to ER stress, indicating RNF186 can arouse stress signaling at ER. We further identified the Bcl-2 family protein BNip1 as one of the substrates of RNF186. BNip1 co-localizes with RNF186 at ER and is poly-ubiquitinated by RNF186 through K29 and K63 linkage in vivo. This modification promotes BNip1 transportation to mitochondria but has no influence on its protein level. The half-life of RNF186 is prolonged under ER stress, probably because of the inhibition on its self-ubiquitination and subsequent degradation by proteasomes. In addition, the ubiquitination of BNip1 is greatly enhanced when ER stress occurred, possibly due to RNF186 accumulation. More importantly, knockdown of BNip1 attenuates the stress signals at ER induced by RNF186. These results collectively indicate that BNip1 functions as a downstream modulator of RNF186 to direct ER stress-associated apoptotic signaling. Our study might reveal a novel E3 ligase-mediated mechanism for modulating ER stress.

RNF152, a novel lysosome localized E3 ligase with pro-apoptotic activities.

RING finger protein 152 (RNF152) is a novel RING finger protein and has not been well characterized. We report here that RNF152 is a canonical RING finger protein and has E3 ligase activity. It is polyubiqitinated partly through Lys-48-linked ubiquitin chains in vivo and this phenomenon is dependent on its RING finger domain and transmembrane domain. RNF152 is localized in lysosomes and co-localized with LAMP3, a lysosome marker. Moreover, over-expression of RNF152 in Hela cells induces apoptosis. These results suggest that RNF152 is a lysosome localized E3 ligase with pro-apoptotic activities. It is the first E3 ligase identified so far that is involved in lysosome-related apoptosis.

[Clinical characteristics and outcomes of respiratory distress syndrome in term and late-preterm neonates].

OBJECTIVE: Respiratory distress syndrome (RDS) is a frequently seen acute respiratory disorder in the newborn infants. Since the original description of deficiency of the pulmonary surfactant in premature neonates by Avery in 1959, RDS has most commonly been attributed to developmental immaturity of surfactant production. But in clinical practice it has been found that there was RDS in term and late-term neonates. Many of them were recognized as transient tachypnea at the beginning, they were diagnosed as RDS until respiratory distress and the typical radiological signs were demonstrated. The purpose of this study was to investigate the clinical characteristics of RDS of term and late-term neonates. METHODS: All neonates admitted to the neonatal intensive care unit of the Children's Hospital, Zhejiang University School of Medicine between May, 2005 and May, 2007 on the basis of RDS were analyzed. RDS was diagnosed when respiratory distress and the typical radiological signs were documented. Patients were grouped into preterm group (Group 1, gestational age < 35 w, n = 103) and term and late-term group (Group 2, gestational age > or = 35 w, n = 74). RESULTS: In Group 1, 76 preterm infants were male, the mean gestational age was 31.1 w, the mean Apgar score at 1 min was 7.6, the mean birth weight was 1702 g, 56 cases were vaginally delivered and 47 were delivered through Cesarean section. Only one was delivered via elective Cesarean section before onset of labor. A total of 88 patients needed mechanical ventilation (MV), the time for beginning MV was 8.7 h (1 - 72 h), and lasted for 4.3 d (0.5 - 29 d). The oxygenation index (OI) was 11.9 (10.00 - 52.63) and PaO2/PAO2 was 0.29 (0.03 - 0.98). Four patients had an OI > 40. A total of 28 patients were treated with pulmonary surfactant (PS), and 11 of the 28 underwent MV, the OI before and 2 h, 8 - 12 h and 20 - 24 h after using PS were 10.5, 5.4, 3.4, and 4.3, respectively (P < 0.01). A total of 33 patients in Group 1 had intracranial hemorrhage, 4 patients had pneumothorax, 4 patients had persistent pulmonary hypertension of the newborn (PPHN) and 15 patients had ventilator associated pneumonia (VAP). In Group 2, 54 infants were male, the mean gestational age was 37 w, the mean Apgar score at 1 min was 8.5, the mean birth weight was 2789 g, 8 cases were vaginally delivered, 66 were delivered through Cesarean section and 59 were delivered via elective Cesarean section before onset of labor. A total of 63 patients needed MV, the time for beginning MV was 27.8 h (1 - 72 h, compared to Group 1, P < 0.01), and lasted for 3.7 d (0.5 - 13.5 d). The OI was 19.70 (10.00 - 56.67, compared to Group 1, P < 0.01) and PaO2/PAO2 was 0.16 (0.017 - 0.470, compared to Group 1, P < 0.01). Seven patients had an OI > 40. A total of 8 patients were treated with PS and 7 of them had MV, the OI before and 2 h, 8 - 12 h and 20 - 24 h after using PS were 11.2, 7.6, 7.5, and 7.6 (the last two compared to group 1, P < 0.01, respectively). A total of 16 patients had pneumothorax, 10 patients had intracranial hemorrhage, 16 patients had PPHN and 7 patients had VAP. CONCLUSION: Most of the term and late-term neonates who developed RDS were delivered through cesarean section before onset of labor. They underwent MV later. The oxygenation was worse than RDS in preterm infants. PS did not have the same effect as seen in preterm infants. They had more pneumothorax and PPHN.

[Application of flow cytometry to detect PP65 antigenemia for diagnosis and monitoring of human cytomegalovirus infection].

OBJECTIVES: To evaluate the clinical significance of flow cytometry (FCM) to detect the cytomegalovirus (CMV) PP65 antigen in patients with CMV infection. METHODS: Samples from 35 patients without CMV infection were used as negative control. The definite diagnosis of CMV infection was based on the national criteria for CMV infection. All 136 patients with CMV infection were examined with the FCM to detect CMV PP65 antigen, real-time fluorescence quantitative-polymerase chain reaction assay (RFQ-PCR) to detect CMV-DNA and ELISA to measure the serum level of IgM antibody against CMV. The results of these 3 assays in 2 groups (isolated organ involvement and disseminated diseases) were compared and the significance of PP65 antigenemia was evaluated. A short-term follow-up was undertaken in 18 patients. RESULTS: The percentages of PP65 positivity in blood mononuclear cells (MNC) and polymorphic nuclear leukocyte (PMNL) from 35 negative control patients were 0.21% +/- 0.09% with a range of 0 - 0.41% and 0.24% +/- 0.10% with a range of 0.12% - 0.48%, respectively, which were not significantly different (t = 0.425, P > 0.05). The 95(th) percentiles (P(95)) of PP65 in MNC and PMNL were 0.39% and 0.45%, respectively, so a cutoff value of >/= 0.50% was set. Of the 136 patients with CMV infection, 118 samples from 118 patients were positive for PP65 antigenemia with a positive rate of 86.8%, which was not statistically different from that (90.4%, chi(2) = 0.91, P > 0.05) of CMV-DNA detected by RFQ-PCR assay but it was significantly higher than that (45.6%, chi(2) = 51.50, P < 0.005) of the detection by IgM measurement. PP65 detection was correlated with urine CMV DNA amplification (chi(2) = 63.78, P < 0.01) while the different detection rates between the two assays were not statistically significant (chi(m)(2) = 1.78,P > 0.05). PP65 detection was not correlated with serum IgM measurement while the detection rates between the two were significantly different (chi(m)(2) = 52.92,P < 0.01). No significant difference was found between the detection rates of CMV infection in MNC (45/53, 84.9%) and PMNL (43/53, 81.1%) (chi(m)(2) = 0.25, P > 0.05). Higher PP65 antigenemia level was correlated with systemic CMV infection, while lower level of PP65 was either in the patients with isolated organ involvement by CMV (chi(2) = 38.51, P < 0.005) or less severe in patient's situation. PP65 antigenemia of CMV infection returned to lower level or negative in recovery stage and increased when condition of patients deteriorated. CONCLUSIONS: PP65 antigenemia detection by FCM is effective in the diagnosis of the active CMV infection. Quantitative monitoring of PP65 antigenemia is useful in the evaluation of patients with CMV infection.