Objective:To investigate the effect of miRNA (miR) -193b-5p on melanogenesis and its possible mechanisms.Methods:Human primary melanocytes were isolated from discarded normal foreskin tissues of healthy males after circumcision, and cultured in vitro. miR-NC mimics (miR-NC mimic group) and miR-193b-5p mimics (miR-193b-5p mimic group) were transfected into human primary melanocytes and human MNT1 melanoma cells, separately. After transfection, real-time quantitative PCR (RT-qPCR) was performed to determine the overexpression efficiency of miR-193b-5p at 48 hours, Western blot analysis to determine the expression of melanogenesis-related proteins tyrosinase (TYR) and microphthalmia-associated transcription factor (MITF) in human primary melanocytes and human MNT1 melanoma cells at 72 hours, and the melanin content in the above cells was determined by a sodium hydroxide solubilization method at 1 week. The target gene of miR-193b-5p was predicted by using Targetscan algorithms and verified by dual-luciferase reporter assay, and RT-qPCR and Western blot analysis were performed to analyze changes in mRNA and protein expression of the target gene respectively after the overexpression of miR-193b-5p. Two-independent-samples t test was used for comparisons between two groups. Results:In human primary melanocytes and human MNT1 melanoma cells, the miR-193b-5p expression levels were significantly higher in the miR-193b-5p mimic groups than in the miR-NC mimic groups ( t = 65.57, 22.49, respectively, both P < 0.001) , and the melanin content was significantly lower in the miR-193b-5p mimic groups (0.091 ± 0.007, 0.130 ± 0.004, respectively) than in the miR-NC mimic groups (0.117 ± 0.002, 0.188 ± 0.032, t = 5.98, 3.24, P < 0.01, < 0.05, respectively) . Western blot analysis showed that the expression of melanogenesis-related proteins TYR and MITF in both human primary melanocytes and human MNT1 melanoma cells was significantly lower in the miR-193b-5p mimic groups than in the miR-NC mimic groups (all P < 0.01) . TargetScan analysis and dual-luciferase reporter assay revealed a binding site for miR-193b-5p in the 3′ untranslated region of the transcriptional regulator CITED2. After up-regulation of miR-193b-5p expression in human primary melanocytes and human MNT1 melanoma cells, the CITED2 mRNA and protein expression levels significantly decreased compared with the miR-NC mimic groups (all P < 0.05) . Conclusion:miR-193b-5p overexpression can down-regulate the expression of melanogenesis-related proteins TYR and MITF, and then inhibit melanogenesis, which may be related to the targeted inhibition of CITED2 expression.

Objective:To understand influencing factors on the deaths of HIV/AIDS patients receiving antireviral treatment in Butuo county of Liangshan Yi Autonomous Prefecture (Liangshan) from 2010 to 2019, to provide data for drug replacement and sustainable antiviral treatment strategy.Methods:A matched case-control study was used to collect basic and follow-up information on AIDS death patients receiving antiviral treatment in Butuo county of Liangshan from 2010 to 2019. The control group was formed by sampling twice the number of cases. The logistic regression model was used to analyze the risk factors affecting mortality.Results:In 3 355 patients of HIV/AIDS treated with antiviral therapy, 1 179 cases in the death group and 2 176 cases in the control group. Including 81.34% were 30-49 years old, 69.09％males, 99.55% Yi nationality, 91.12% were married or cohabitated, 95.77% had junior high school education or below, and 88.41% peasants. Amultivariate logistic stepwise regression model showed that among the death risk factors, age ≥50 years old was 5.08 times (95% CI:3.05-8.48) that of the 18-29, female was 0.70 times (95% CI: 0.52-0.94) than male, the transmission rate of intravenous drug use was 1.43 times (95% CI: 1.06-1.91) that of heterosexual transmission, CD4 +T lymphocyte (CD4) count ≥350 cells/μl before treatment was 0.38 times (95% CI: 0.30-0.48) that of CD4 <200 cells/μl before treatment, the most recent antiviral treatment regimen containing LPV/r was 0.04 times (95% CI: 0.01-0.18) than that of stavudine (d4T) + lamivudine (3TC) + nevirapine (NVP)/efavirenz (EFV) regimen, drug resistance was 3.40 times (95% CI: 2.13-5.42) of non-drug resistance, non-viral load and non-drug resistance test results were 12.98 times (95% CI: 10.28-16.40) of non-drug resistance. Conclusions:Age, gender, transmission route, CD4 before treatment, the latest antiviral treatment program, and drug resistance test after antiviral therapy were the influencing factors of HIV/AIDS death in Butuo county. It is necessary to expand the coverage of viral load and drug resistance test to change the antiviral therapeutic schedule scientifically and carry out publicity and education on the compliance of patients with antiviral treatment and medical staff training in order to reduce the mortality of patients with antiviral treatment.

Objective To investigate the distribution and drug resistance of carbapenem-resistant Enterobacteriaceae ( CRE) isolated from children in China. Methods CRE strains were collected in 10 ter-tiary children's hospitals of China from January 1, 2016 to December 31, 2017. Antimicrobial susceptibility of the clinical strains was detected with disk diffusion method ( KB method) and automated method. The re-sults were analyzed according to the Clinical and Laboratory Standards Institute ( CLSI) Standards published in 2017. WHONET 5. 6 software was used to retrospectively analyze the distribution characteristics and drug resistance of these strains. Results A total of 3065 CRE clinical strains were isolated from children with an overall prevalence of 7. 7％ and among them, 13. 5％ were isolated in neonatal group and 5. 8％ in non-neo-natal group. The detection rate of CRE in 2017 was higher than that in 2016 (9. 7％ vs 5. 7％). Among the 3065 CRE strains, there were 1912 strains of Klebsiella pneumoniae (62. 0％), 667 strains of Escherichia coli (22. 0％), 206 strains of Enterobacter cloacae (7. 0％), 56 strains of Klebsiella aerogenes (1. 8％) and 47 strains of Serratia marcescens (1. 5％). Most of the strains were isolate in neonatology departments including neonatal intensive care units (NICU) and intensive care units (ICU), accounting for 44. 8％ and 19. 7％, respectively. Respiratory tract (61. 8％), urine (19. 4％) and blood (5. 7％) specimens were the main sources of CRE isolates. Results of antimicrobial susceptibility test showed that the CRE strains were highly resistant to carbapenem antibiotics such as imipenem, meropenem and ertapenem, as well as penicillins and most cephalosporins (79. 6％-100％), especially those isolated in the neonatal group (P<0. 05). Children had relatively low resistance rates to aminoglycosides such as amikacin (19. 7％) and fos-fomycin (11. 9％), fluoroquinolones such as levofloxacin (37. 7％) and ciprofloxacin (43. 3％), and tige-cycline (3. 8％). Currently, no polymyxin B-resistant strains were isolated. Conclusions The prevalence of common CRE strains in children in 2017 was higher than that in 2016, especially in newborns. Drug re-sistance in CRE strains isolated from neonates to common antibiotics was more severe, suggesting that great attention should be paid to it and timely measures should also be taken.

Objective: To analyze the antimicrobial resistance profile in Chinese children. Methods: This was a prevalence survey. From January 1 through December 31, 2016, the isolates were collected from 10 tertiary children hospitals in China. Antimicrobial susceptibility testing was carried out by routine laboratory methods. The penicillin susceptibility of streptococcus pneumonia and Meropenem susceptibility of gram-negative bacteria were detected by E-test and disk diffusion method respectively. Antimicrobial susceptibility results were interpreted according to the criteria of Clinical and Laboratory Standards Institute (CLSI) Guideline 2016. The data of antimicrobial susceptibility testing of isolates from either the different patients (neonatal group and non-neonatal group) or various sources were analyzed by WHONET 5.6 software. Results: A total of 56 241 isolates were collected, of which 41.5% (23 328 isolates) were gram-positive organisms and 58.5% (32 886 isolates) gram-negative organisms. The five leading pathogens were Escherichia coli (7 995/56 214, 14.2%), Straphylococcus aureus (6 468/56 214, 11.5%), Streptococcus pneumonia (6 225/56 214, 11.1%), Haemophilus influenza (5 435/56 214, 9.7%) and Klebsiella pneumonia (4 523/56 214, 8.0%). The Meropenem resistance rates of Klebsiella pneumonia, Enterobacter cloacae, Escherichia coil, Pseudomonas aeruginosa, Acinetobacter baumonia isolates were 27.4% (326/1 189) , 8.1% (29/358) , 2.0% (27/1 362) , 19.5% (34/174) , 49.7% (230/463) in neonatal group and 15.4% (512/3 327) , 4.8% (40/841) , 2.3% (151/6 564) , 13.7% (252/1 840) , and 53.4% (860/1 611) in non-neonatal group. The Methicillin-resistant Staphylococcus aureus (MRSA) rates of neonatal group and non-neonatal group were 46.2% (649/1 404) and 33.3% (1 668/5 010) . The penicillin non-susceptible rates of Streptococcus pneumonia in the two groups were 17.6% (6/34) and 18.2% (1 121/6 158) respectively. The β-lactamase positive rates of Haemophilus pneumonia isolates in the neonatal group and non-neonatal groups were 33.8% (47/139) and 44.4% (2 345/5 282) respectively. Conclusion This investigation highlights the worrisome trend of antimicrobial resistance in children, especially among neonatal patients in China.

Objective: To describe the clinical characteristics of pneumococcal infections and drug resistance of Streptococcus pneumoniae isolates from children's hospitals, which would provide reference for preventing and treating pneumococcal diseases. Methods: This was a prevalence survey. In this study, the age, specimen type, monthly distribution characteristics, and antimicrobial resistance of Streptococcus pneumoniae isolates from 9 children's hospitals in China were investigated between January 1, 2016 and December 31, 2016. The WHONET 5.6 software was used to analyze the antibiotic susceptibility of Streptococcus pneumoniae. The comparison of rates was performed by Chi-square test. Results: A total of 6 200 isolates of streptococcus pneumoniae were obtained, namely, 95.1% (5 876/6 177) from the respiratory tract specimens, 2.2% (136/6 177) from blood specimens and 0.4% (24/6 177) from cerebrospinal fluid specimens. The isolates were mainly from children older than 1 and younger than 5 years (54.7%, 3 381/6 185) . Most of strains (33.2%, 1 184/3 563) were isolated in November, December and January. Streptococcus pneumoniae isolates were completely sensitive to vancomycin (100.0%, 6 189/6 189) , linezolid (100.0%, 6 030/6 030) , moxifloxacin (100.0%, 3 064/3 064) , highly sensitive to levofloxacin (99.8%, 5 528/5 540), ertapenem (98.8%, 3 024/3 061) and lowly sensitive to erythromycin (1.7%, 102/6 016), clindamycin (3.7%, 116/3 136), and tetracycline (5%, 244/4 877), respectively. According to the parenteral susceptibility breakpoints for non-meningitis isolates, the sensitivity of Streptocococus pneumoniae to penicillin from children's hospital of Chongqing Medical University (49.3%, 892/1 809) was significantly lower than those of other hospitals (χ²=1 268.161, P<0.05) . Conclusions Streptococcus pneumoniae is mainly isolated from respiratory tract, from children older than 1 and younger than 5 years and during November to January in tertiary children's hospital of China. The Streptococcus pneumoniae from children is highly sensitive to vancomycin, linezolid, moxifloxacin, levofloxacin. There are also significant differences in the sensitivity of penicillin for Streptococcus pneumoniae from different hospitals.

OBJECTIVETo detect pathogenic mutation of the SLC39A4 gene in a male patient with acrodermatitis enteropathica (AE).

METHODSPeripheral venous blood sample and clinical data from the patient and his parents were collected. One hundred unrelated healthy individuals were recruited as controls. All coding exons and flanking exon-intron sequences of the SLC39A4 gene were analyzed by PCR and direct sequencing.

RESULTSThe results revealed that the patient and his mother have both carried a novel frame-shift mutation c.1110InsG (p.Gly370GlyfsX47 to TGA) in exon 6. A novel nonsense mutation c.958C to T (p.Q320X) in exon 5 was also detected in the patient and his father and grandmother. This novel mutation was not detected in the unaffected family members and 100 unrelated healthy controls.

CONCLUSIONThe novel frame-shift mutation c.1110InsG (p.Gly370GlyfsX47 to TGA) derived from the mother and nonsense mutation c.958C to T (p.Q320X) of the SLC39A4 gene derived from the father may underlie the disease in the patient.

Acrodermatitis ; genetics ; Adolescent ; Base Sequence ; Cation Transport Proteins ; genetics ; Exons ; Homozygote ; Humans ; Male ; Molecular Sequence Data ; Mutation ; Pedigree ; Zinc ; deficiency

OBJECTIVETo identify pathogenic mutations of TSC1 and TSC2 genes in two familial and one sporadic cases with tuberous sclerosis complex (TSC).

METHODSFor five patients and their family members, potential mutations of the TSC1 and TSC2 genes were detected by direct sequencing.

RESULTSFor one family, a novel missense mutation c.1964C>T (p.S655F) was detected in the exon 19 of the TSC2 gene. For the sporadic patient, a repeat substitution with deletion mutation c.5238-5255delCATCAAGCGGCTCCGCCA (p.His1746GlnfsX56) was detected in the exon 40 of the TSC2 gene, which led to a stop codon TGA after the 56th amino acids. No mutation was found in another family.

CONCLUSIONThe missense mutation c.1964C>T(P.S655F) and the substitution with deletion mutation 5238-5255delCATCAAGCGGCTCCGCCA(p.His1746GlnfsX56) of the TSC2 gene probably underlie the disease in the first family and the sporadic case.

Adolescent ; Adult ; Base Sequence ; Child, Preschool ; DNA Mutational Analysis ; Female ; Humans ; Male ; Mutation, Missense ; Pedigree ; Phenotype ; Tuberous Sclerosis ; genetics ; Tumor Suppressor Proteins ; genetics

ObjectiveTo explore the effects of social isolation on the cognition and expression of 5-HT2C receptor(5-HT2CR) and adenosine deaminase that act on RNA 1(ADAR1) in BALB/c mice.MethodsThe healthy BALB/c mice were isolated for 2,4,and 8 weeks individually since postnatal 21 days respectively to set up isolation mice model,the same age mice without isolation were regarded as control group.The new object location and the new object recognition tests were used to measure the spatial and non-spatial cognitive function,and western blot was used to measure the protein expression of 5-HT2CR and ADAR1.ResultsThe new object location test showed that the spatial discrimination index (DI) of BALB/c mice isolated for 2 weeks was decreased significantly compared with the control group(control group was (0.075±0.340),isolation group was (-0.653±0.308),P<0.05),and no obvious difference was found for the group isolated for 4 and 8 weeks.The new object recognition test showed that the non-spatial DI of BALB/c mice isolated for 2 and 4 weeks were decreased significantly compared with the control group(control 2 weeks group was (0.088±0.210),isolation 2 weeks group was (-0.945±0.194),P<0.05;control 4 weeks group was (0.105±0.267),isolation 4 weeks group was (-0.506±0.215),P<0.05),and no obvious difference was found for the group isolated for 8 weeks.Compared with the control group the expression of 5-HT2CR and ADAR1 in the hippocampus were decreased significantly for the group isolated for 2 weeks.(5-HT2CR:control group was (1.025±0.144),isolation group was (0.891±0.026),P<0.05.ADAR1: control group was (0.839±0.120),isolation group was (0.629±0.094),P<0.05).ConclusionsTwo week social isolation results in the decrease of spatial and non-spatial cognitive function in BALB/c mice,in the meanwhile,social isolation stress results in the obvious decrease of 5-HT2C receptor and ADAR1 protein expression in the hippocampus of BALB/c mice.

Objective To observe the action ofFu Liu Zhen (acupuncture at the six abdominal points) plus moxa-box moxibustion at umbilicus on a community population of qi and yang deficiency.Method Two hundred subjects diagnosed with qi deficiency and/or yang deficiency were randomized into a treatment group and a control group, 100 cases in each group. The two groups both received constitution-associated health education, and the treatment group was additionally intervened byFu Liu Zhen plus moxa-box moxibustion at umbilicus, while the control group didn’t receive any other intervention. The Score of Constitution in Traditional Chinese Medicine (TCM) and the number of people of each constitution were observed prior to the intervention, right after the intervention, 6 months after the intervention and 12 months after the intervention.Result In the treatment group, the scores of qi deficiency and yang deficiency after the intervention, 6 months after the intervention and 12 months after the intervention were significantly different from that prior to the intervention (P<0.01). The scores of the corresponding constitution types (qi deficiency, yang deficiency) after the intervention, 6 months after the intervention and 12 months after the intervention were significantly different from that before the intervention (P<0.01). The proportions of subjects with harmonious constitution, qi deficiency, and yang deficiency in the treatment group after the intervention, 6 months and 12 months after the intervention were significantly different from that prior to the intervention (P<0.01). The proportions of subjects with harmonious constitution, qi deficiency, and yang deficiency in the treatment group after the intervention, 6 months and 12 months after the intervention were significantly different from that in the control group (P<0.01).ConclusionFu Liu Zhen plus moxa-box moxibustion at umbilicus can improve the qi-deficiency and yang-deficiency constitutions in the community population.

OBJECTIVETo identify potential mutation of the ADAR1 gene in a Chinese family and a sporadic case affected with dyschromatosis symmetrica hereditaria(DSH).

METHODSClinical data and peripheral blood samples from the pedigree and the sporadic patient were collected. Following extraction of genomic DNA, all 15 exons and exon-intron flanking sequences of the ADAR1 gene were amplified by polymerase chain reaction and subjected to direct sequencing.

RESULTSA novel frame-shift mutation c.2638delG (p.Asp880ThrfsX15) from the patients of the pedigree was detected in exon 8 of the ADAR1 gene. And a novel nonsense mutation c.2867C>A (p.Ser956X) was detected in exon 10 of the ADAR1 gene from the sporadic case. Neither mutation was identified among the unaffected family members nor 100 unrelated healthy controls.

CONCLUSIONThe frame-shift mutation c.2638delG (p.Asp880ThrfsX15) and the nonsense mutation c.2867C>A (p.Ser956X) in the ADAR1 gene probably underlie the DSH in our patients.

Adenosine Deaminase ; genetics ; Adult ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; China ; Codon, Nonsense ; Exons ; Female ; Frameshift Mutation ; Humans ; Male ; Molecular Sequence Data ; Pedigree ; Pigmentation Disorders ; congenital ; enzymology ; genetics ; RNA-Binding Proteins ; genetics