Objective To investigate the effects of different methods of preparation on the contents of nuzhenide and oleanolic acid in Ligustri Lucidi Fructus. Methods The same batch of Ligustri Lucidi Fructus was under preparation and process. RP-HPLC was used to determine the contents of nuzhenide and oleanolic acid in crude samples, wine-prepared samples, vinegar-prepared samples, and salt-prepared samples. Results The contents of nuzhenide were in the order as follow:salt-prepared samples (1.997 5%)>crude samples (1.600 7%)>wine-prepared samples (1.144 6%)>vinegar-prepared samples (0.894 6%). The contents of oleanolic acid were in the order as follow:wine-prepared samples (1.120 2%)>salt-prepared samples (0.924 6%)>vinegar-prepared samples (0.913 2%)>crude samples (0.899 8%). Conclusion Different methods of preparation have certain effects on the contents of nuzhenide and oleanolic acid in Ligustri Lucidi Fructus.

Objective To compare the efficacy and complications between transurethral resec-tion of the prostate(TURP) and transurethral vaporization-resection of the prostate (TUVRP). Methods 637 cases of benign prostatic hyperplasia(BPH)were divided into 2 groups, 298 cases un-derwent TURP and 339 underwent TUVRP. The maximal flow rates (Qmax) were (9.8±2.3)ml/s, (10.1±2.1) ml/s, the international prostatic symptom scores (IPSS) were 15.3±3.1,15.1±3.7 re-spectively. The surgical outcomes and the complications of the 2 groups were analyzed. Results For the TURP group and TUVRP group, the postoperative Qmax were (19.0±2.9)ml/s and (18.0±2.3) ml/s, both significantly higher than those of preoperation(P<0.01). There was no significant differ-ence between the 2 groups(P0.05). For the TURP group, the mean operation time was (52±16) rain, visual hematuria lasting for (9.0±2.3) d, secondary bleeding in 6 cases (2.0%), lower urinary tract infection in 14 cases(4.7%), 1 month after operation lower urinary tract symptom(LUTS) in 26 case(8.7%), IPSS was 5.0±1.4, contemporary incontinence in 6 cases(2.0%), memberanous sticture 4 cases(1.3%). For the TUVRP group, the mean operation time was (68%19)min, visual hematuria lasting for (12.0±3.6) d, secondary bleeding in 19 cases (5.6%), lower urinary tract infection in 38 cases(11.2%) ,1 month after operation LUTS in 59 cases(17.4%) ,IPSS was 8.0±1.6,contemporary incontinence in 13 cases(3.8%), memberanous stieture in 16 cases (4.7%). There were significant differences between the 2 groups. For the TURP group, blood loss during operation was (126±29) ml, resected tissue weighed (31±8)g, pre- and post-operative serum natium consentration balance was (8±6)mmol/L,TURS developed in 3 cases(1%). For the TUVRP group, blood loss during opera-tion was (122±38)ml, resected tissue weighed (33±9)g,pre- and post-operative serum natium con-sentration balance was (7±7) mmol/L, TURS developed in 2 cases(0.6%). There was no significant differences(P0.05). Conclusions TURP and TUVRP have similar efficacy treating symptomatic BPH. The incidences of complications of TUVRP are slightly higher than those of TURP.

IgA nephropathy is the most common form of primary glomerulonephritis which mainly accounts for the development of end-stage renal diseases. It is characterized by deposits of IgA1 in mesangium. The pathogenesis of IgA nephropathy is complicated. Moreover, there is a wide range of clinical features and variable histomorphologies in the diagnosed cases of IgA nephropathy. It was demonstrated that the galactose-deficient of IgA1 O-glycan chains led IgA1 to self-aggregation and eventual deposition in mesangium. Abnormality of glycosyltransferases, genetic mutation and immunologic disorder were involved in the aberrant glycosylation of IgA1 which was recognized as the key etiopathogenisis of IgA nephropathy. However, the exact source and the pathogenic mechanism of aberrantly glycosylated IgA1 remain obscure. The further studies on aberrant O-glycosylation of IgA1 would contribute to the understanding of IgA nephropathy and provide new therapeutical strategy.
Animals ; Glomerulonephritis, IGA ; etiology ; metabolism ; Glycosylation ; Humans ; Immunoglobulin A ; metabolism

VC2002, isolated from postweaning multisystemic wasting syndrome (PMWS)-affected pig, is a mixture of two porcine circovirus genotype 2b (PCV2b) viruses, K2 and K39. Preliminary experiments disclosed short-term adverse effects of K39, but not K2, on porcine foetuses. These findings led to the hypothesis that infection of immuno-incompetent foetuses with K2 confers a status of immunotolerance, and postnatal super-infection with K39 triggers PMWS. To explore this hypothesis, nine 55-day-old foetuses were inoculated in utero (three with K2-104.3TCID50, three with K39-104.3TCID50 and three with medium), and foeto-pathogenicity examined. At 21 days post-inoculation (dpi), K2 did not induce pathology, whereas pathological effects of K39 were evident. Twenty-four 45-day-old foetuses were subsequently inoculated to examine the long-term effect of K2, including six with K2-high dose-104.3TCID50, six with K2-low dose-102.3TCID50 and 12 mock-inoculated controls. Both doses resulted in ifve mummiifed foetuses and one live-born piglet each (69dpi). K2 was recovered from all mummies. K2 and K2-speciifc antibodies were not detected in serum of the two live-born piglets at birth, indicating full control of K2 infection. The K2-low dose-infected piglet was immunostimulated at day 2, but not the K2-high dose-infected piglet. Both non-stimulated and stimulated K2-infected piglets were super-inoculated with K39 at day 6 or 8 (taken as 0 days post super-inoculation). Low viral replication was observed in the non-stimulated K2-K39 piglet (up to 103.3 TCID50/g;identiifed as K39). In contrast, viral replication was extremely high in the stimulated K2-K39 piglet (up to 105.6TCID50/g) and identiifed as K2, indicating that K2 infection is controlled during foetal life, but emerges after birth upon immunostimulation. However, none of the piglets showed any signs of PMWS.

BACKGROUND/AIMS: Neurotensin is a gut-brain peptide with both inhibitory and excitatory actions on the colonic musculature; our objective was to understand the implications of this for motor patterns occurring in the intact colon of the rat. METHODS: The effects of neurotensin with concentrations ranging from 0.1-100 nM were studied in the intact rat colon in vitro, by investigating spatio-temporal maps created from video recordings of colonic motility before and after neurotensin. RESULTS: Low concentration of neurotensin (0.1-1 nM) inhibited propagating long distance contractions and rhythmic propagating motor complexes; in its place a slow propagating rhythmic segmental motor pattern developed. The neurotensin receptor 1 antagonist SR-48692 prevented the development of the segmental motor pattern. Higher concentrations of neurotensin (10 nM and 100 nM) were capable of restoring long distance contraction activity and inhibiting the segmental activity. The slow propagating segmental contraction showed a rhythmic contraction—relaxation cycle at the slow wave frequency originating from the interstitial cells of Cajal associated with the myenteric plexus pacemaker. High concentrations given without prior additions of low concentrations did not evoke the segmental motor pattern. These actions occurred when neurotensin was given in the bath solution or intraluminally. The segmental motor pattern evoked by neurotensin was inhibited by the neural conduction blocker lidocaine. CONCLUSIONS: Neurotensin (0.1-1 nM) inhibits the dominant propulsive motor patterns of the colon and a distinct motor pattern of rhythmic slow propagating segmental contractions develops. This motor pattern has the hallmarks of haustral boundary contractions.
Absorption ; Animals ; Baths ; Colon* ; In Vitro Techniques ; Interstitial Cells of Cajal ; Lidocaine ; Myenteric Plexus ; Neural Conduction ; Neurotensin* ; Peristalsis ; Rats* ; Receptors, Neurotensin ; Video Recording

INTRODUCTION: Frontline healthcare workers (HCWs) exposed to coronavirus disease 2019 (COVID-19) are at risk of psychological distress. This study evaluates the psychological impact of COVID-19 pandemic on HCWs in a national paediatric referral centre. METHODS: This was a survey-based study that collected demographic, work environment and mental health data from paediatric HCWs in the emergency, intensive care and infectious disease units. Psychological impact was measured using the Depression, Anxiety, Stress Scale-21. Multivariate regression analysis was performed to identify risk factors associated with psychological distress. RESULTS: The survey achieved a response rate of 93.9% (430 of 458). Of the 430 respondents, symptoms of depression, anxiety and stress were reported in 168 (39.1%), 205 (47.7%) and 106 (24.7%), respectively. Depression was reported in the mild (47, 10.9%), moderate (76, 17.7%), severe (23, 5.3%) and extremely severe (22, 5.1%) categories. Anxiety (205, 47.7%) and stress (106, 24.7%) were reported in the mild category only. Collectively, regression analysis identified female sex, a perceived lack of choice in work scope/environment, lack of protection from COVID-19, lack of access to physical activities and rest, the need to perform additional tasks, and the experience of stigma from the community as risk factors for poor psychological outcome. CONCLUSION A high prevalence of depression, anxiety and stress was reported among frontline paediatric HCWs during the COVID-19 pandemic. Personal psychoneuroimmunity and organisational prevention measures can be implemented to lessen psychiatric symptoms. At the national level, involving mental health professionals to plan and coordinate psychological intervention for the country should be considered.
Adult ; Anxiety/etiology* ; COVID-19/psychology* ; Depression/etiology* ; Female ; Health Surveys ; Hospitals, Pediatric ; Humans ; Logistic Models ; Male ; Middle Aged ; Multivariate Analysis ; Occupational Diseases/etiology* ; Pandemics ; Personnel, Hospital/psychology* ; Prevalence ; Risk Factors ; Self Report ; Severity of Illness Index ; Singapore/epidemiology* ; Stress, Psychological/etiology*

To clone the first anion channel from Xenopus laevis (X. laevis), we isolated a calcium-activated chloride channel (CLCA)-like membrane protein 6 gene (CMP6) in X. laevis. As a first step in gene isolation, an expressed sequence tags database was screened to find the partial cDNA fragment. A putative partial cDNA sequence was obtained by comparison with rat CLCAs identified in our laboratory. First stranded cDNA was synthesized by reverse transcription polymerase-chain reaction (RT-PCR) using a specific primer designed for the target cDNA. Repeating the 5' and 3' rapid amplification of cDNA ends, full-length cDNA was constructed from the cDNA pool. The full-length CMP6 cDNA completed via 5'- and 3'-RACE was 2,940 bp long and had an open reading frame (ORF) of 940 amino acids. The predicted 940 polypeptides have four major transmembrane domains and showed about 50% identity with that of rat brain CLCAs in our previously published data. Semi-quantification analysis revealed that CMP6 was most abundantly expressed in small intestine, colon and liver. However, all tissues except small intestine, colon and liver had undetectable levels. This result became more credible after we did real-time PCR quantification for the target gene. In view of all CLCA studies focused on human or murine channels, this finding suggests a hypothetical protein as an ion channel, an X. laevis CLCA.
Amino Acids ; Animals ; Brain ; Chloride Channels ; Clone Cells ; Colon ; DNA, Complementary ; Expressed Sequence Tags ; Humans ; Intestine, Small ; Ion Channels ; Liver ; Membrane Proteins ; Membranes ; Open Reading Frames ; Peptides ; Rats ; Real-Time Polymerase Chain Reaction ; Resin Cements ; Reverse Transcription ; Staphylococcal Protein A ; Tissue Distribution ; Xenopus ; Xenopus laevis

OBJECTIVE: Clinical presentation and physical signs may be unreliable in the diagnosis of stercoral colitis (SC). This study evaluates the value of computed tomography (CT) in distinguishing fatal from non-fatal SC. MATERIALS AND METHODS: Ten patients diagnosed as SC were obtained from inter-specialist conferences. Additional 13 patients with suspected SC were identified via the Radiology Information System (RIS). These patients were divided into two groups; fatal and non-fatal SCs. Their CT images are reviewed by two board-certified radiologists blinded to the clinical data and radiographic reports. RESULTS: SC occurred in older patients and displayed no gender predisposition. There was significant correlation between fatal SC and CT findings of dense mucosa (p = 0.017), perfusion defects (p = 0.026), ascites (p = 0.023), or abnormal gas (p = 0.033). The sensitivity, specificity, and accuracy of dense mucosa were 71%, 86%, and 81%, respectively. These figures were 75%, 79%, and 77% for perfusion defects; 75%, 80%, and 78% for ascites; and 50%, 93%, and 78% for abnormal gas, respectively. Each CT sign of mucosal sloughing and pericolonic abscess displayed high specificity of 100% and 93% for diagnosing fatal SC, respectively. However, this did not reach statistical significance in diagnosing fatal SC. CONCLUSION: CT appears to be valuable in discriminating fatal from non-fatal SC.
Adult ; Aged ; Aged, 80 and over ; Chi-Square Distribution ; Colitis/mortality/*radiography ; Contrast Media/diagnostic use ; Diagnosis, Differential ; Fecal Impaction/mortality/*radiography ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Risk Factors ; Sensitivity and Specificity ; Statistics, Nonparametric ; Tomography, X-Ray Computed/*methods

Human pluripotent stem cells (hPSCs), including embryonic stem cells (ESCs) and induced PSCs (iPSCs), represent potentially unlimited cell sources for clinical applications. Previous studies have suggested that hPSCs may benefit from immune privilege and limited immunogenicity, as reflected by the reduced expression of major histocompatibility complex class-related molecules. Here we investigated the global immune-related gene expression profiles of human ESCs, hiPSCs and somatic cells and identified candidate immune-related genes that may alter their immunogenicity. The expression levels of global immune-related genes were determined by comparing undifferentiated and differentiated stem cells and three types of human somatic cells: dermal papilla cells, ovarian granulosa cells and foreskin fibroblast cells. We identified the differentially expressed genes CD24, GATA3, PROM1, THBS2, LY96, IFIT3, CXCR4, IL1R1, FGFR3, IDO1 and KDR, which overlapped with selected immune-related gene lists. In further analyses, mammalian target of rapamycin complex (mTOR) signaling was investigated in the differentiated stem cells following treatment with rapamycin and lentiviral transduction with specific short-hairpin RNAs. We found that the inhibition of mTOR signal pathways significantly downregulated the immunogenicity of differentiated stem cells. We also tested the immune responses induced in differentiated stem cells by mixed lymphocyte reactions. We found that CD24- and GATA3-deficient differentiated stem cells including neural lineage cells had limited abilities to activate human lymphocytes. By analyzing the transcriptome signature of immune-related genes, we observed a tendency of the hPSCs to differentiate toward an immune cell phenotype. Taken together, these data identify candidate immune-related genes that might constitute valuable targets for clinical applications.
Embryonic Stem Cells ; Female ; Fibroblasts ; Foreskin ; Granulosa Cells ; Humans* ; Induced Pluripotent Stem Cells ; Lymphocyte Culture Test, Mixed ; Lymphocytes* ; Major Histocompatibility Complex ; Phenotype ; Pluripotent Stem Cells* ; RNA ; Signal Transduction ; Sirolimus ; Stem Cells ; Transcriptome

BACKGROUNDVon Hippel-Lindau disease (VHL), a heritable autosomal dominant disease characterized by neoplasia in multiple organ systems, has rarely been reported in Asia. We genetically investigated a unique Chinese family with VHL disease and performed an analysis of the VHL protein stability.

METHODSGenomic deoxyribonucleic acid (DNA) extracted from peripheral blood was amplified by polymerase chain reaction (PCR) to three exons of the VHL gene in 9 members of the Chinese family with VHL disease. PCR products were directly sequenced. We estimated the effects of VHL gene mutation on the stability of pVHL, which is indicated by the free energy difference between the wild-type and the mutant protein (ΔΔG).

RESULTSThe Chinese family was classified as VHL type 1. Three family members, including two patients and a carrier, had a T to G heterozygotic missense mutation at nucleotide 515 of the VHL gene exon 1. This missense mutation resulted in the transition from leucine to arginine in amino acid 101 of the VHL protein. There was low stability of the VHL protein (the ΔΔG was 12.71 kcal/mol) caused by this missense mutation.

CONCLUSIONSWe first reported a family with this VHL gene mutation in Asia. This missense mutation is predicted to significantly reduce the stability of the VHL protein and contribute to the development of the renal cell carcinoma (RCC) phenotype displayed by this family. The genetic characterization and protein stability analysis of families with VHL disease are important for early diagnosis and prevention of the disease being passed on to their offspring.

Adult ; China ; Female ; Humans ; Male ; Middle Aged ; Mutation, Missense ; Protein Stability ; Von Hippel-Lindau Tumor Suppressor Protein ; chemistry ; genetics ; von Hippel-Lindau Disease ; genetics