Let-7 is one of the most widely studied in miRNAs at present. let-7 can inhibit cell proliferation, promote cell differentiation and apoptosis, etc. Reduced expression of let-7 in different human tumor tissues or cells is accompanied by the changes of its target genes expression, which suggests that let-7 is closely linked to cancer.

Objective To establish TaqMan probe real‐time Fluorescent Quantitative PCR in detecting Cryptococcus neoformans genomic DNA ,and to provide important method for detection of cryptococcal meningitis .Methods According to the Cryptococcus neoformans ITS‐rDNA sequences obtained from NCBI ,specific primers and probe were designed based on the conserved sequences , a specific 114 bp fragment was amplified by primers and probe ,then recombinant plasmid was constructed .Eight different concen‐trations from 1 .42 × 10 copy/μL to 1 .42 × 10 copy/μL were used as templates by 2 μL .In the optimum reaction condition ,the sen‐sitivity ,specificity and repeatability were evaluated and standard curve was established .15 clinical cryptococcal meningitis strains i‐solated from clinical diagnosis patients were detected .Results The real‐time PCR showed high sensitivity and specificity and was a‐ble to detect 2 .84 × 102 copies of plasmid DNA .The detection sensitivity was 2 .84 × 102 copies plasmid DNA by real‐time PCR ,no amplification curve was detected with human genomic DNA ,other fungus ,bacterias and viruses .The CV of inter‐assay were 2 .86% ,1 .48% and 1 .36% respectively with excellent reproducibility .Fifteen clinical isolated strains could be detected accurately . Conclusion A method of detection of Cryptococcus neoformans DNA by real‐time PCR is established successfully with high sensi‐tivity and specificity ,and the results are stable ,could diagnose cryptococcal meningitis rapidly and early .

Objective To investigate the effects of vitamin K1 in the adjuvant treatment of spasmodic cough in pertussis syndrome children,and its impacts on length of hospital stay.Methods 87 pertussis syndrome children were randomly divided into two groups,among which 41 cases in the control group were treated with the therapies such as anti -infection,respiratory support,antispasmodic,expectorant et al,while 46 cases in the treatment group were treated additionally with vitamin K1 on the basis of the control group.Then compared the duration and hospital stay of the two groups with spasmodic cough.Results The disappeared time of spasmodic cough in the treatment group was (7.22 ±1.33)d and hospital stay was (9.52 ±1.84)d,which in the control group were (15.51 ±2.73)d,(18.71 ± 3.30)d respectively,and there were statistically significant differences(t =-18.310,-16.269,all P <0.01).In the treatment group,the total effective rate was 93.5%,there were 18 cases with markedly effective,25 cases with effective and 3 cases with ineffective.Correspondingly,there were 7 cases markedly effective,23 cases effective, 11 cases ineffective,and total efficiency rate was only 73.1% in the control group,the difference was statistically significant(χ2 =4.285,P <0.05 ).Conclusion Vitamin K1 to alleviate spasmodic cough in children has a significant effect,which can reduce the duration of the cough time and hospital stay in pertussis syndrome children, and also can improve the clinical effect.

Objective:To study short-term effects of PM10(particulate matter of aerodynamic diameter

Objective To study the hemodynamic characteristics of ductus venosus,and its relationship to inflammatory cytokines in ovine fetus with systemic inflammatory response syndrome.Methods Ten near-term fetal sheep radomly divided into two groups.Five of them were in experimental group,and five of them were in control group.All of the animals underwent abdominal cordocentesis guided by ultrasound at the term of 120-130 days,with a LPS injection at a dosage of 10 μg/kg of fetal weight in experimental group,and the same quantity of 0.9% NaCl solution in control.Doppler echocardiography were performed to determine hemodynamics changes of ductus venosus at 0.5 hour before the LPS injection,and at 1 hour,3 hours,6 hours after LPS injection.Meanwhile,fetal umbilical vein blood was sampled for ELISA essay of serum TNF-α and IL-6 at each of the above time points.Results In experimental group,the ductus venosus waveform PI values(DVPI),maximum velocity during cardiac ventricle systole(S)and during cardiac ventricle diastole(D),ductus venosus index(DVI),ductus venosus blood flow(DVQ),the ratio of S and ductus venous maximum velocity during atrial contracton(S/A),(S-A)/D increased with the time after LPS injection,and still significantly increased as compared with the control animals(all P＜0.05).A decreased with the time after LPS injection,and still significantly changed as compared with the controls(P＜0.05).And the ductus venosus diameter(DVD)had not obvious change(P＞0.05).There were significantly positive correlations between DVPI and TNF-α,IL-6(all P＜0.05),negative corelations between A and TNF-α,IL-6(all P＜0.05),and positive correlations between S wave,DVI,DVQ,D wave,S/A,(S-A)/D and IL-6(all P＜0.05),but not obvious relationships between S wave,DVI,DVQ,D wave,S/A,(S-A)/D and TNF-a(all P＞0.05).Condusions DVPI and A might be easy and useful quantitative parameters in the evaluation of fetal systemic inflammatory respome syndrome with LPS injection.

Objective To explore the relationship between glucose-6-phosphate dehydrogenase deficiency and type 2 diabetes and to provide reference for clinical diagnosis,monitoring and treatment of Type 2 diabetes.Methods Subjects were selected from our hospital,including 173 cases of healthy volunteers assigned to the control group;1 95 cases with type 2 diabetes conforming to the diagnostic criteria of WHO were assigned to the diabetic group.97 cases were diagnosed with glucose-6-phosphate dehydrogenase deficiency in our hospital,of whom 82 cases were assigned to simple glucose-6-phosphate dehydrogenase deficiency group,and 1 7 cases were assigned to the diabetes with glucose-6-phosphate dehydrogenase deficiency group.The correlation of glucose-6-phos-phate dehydrogenase activity and diabetes was measured by each detected value.Results Compared with the control group,the glu-cose-6-phosphate dehydrogenase activity and RBC count in diabetic group were higher(P <0.05).Positive correlations between glu-cose-6-phosphate dehydrogenase activity and RBC count in the two groups were significant(P <0.05).HbA1c and FBG showed a significant positive correlation in the control group,diabetic group and diabeties with glucose-6-phosphate dehydrogenase deficiency group(P <0.05).But there was no significant correlation in the glucose-6-phosphate dehydrogenase deficiency group.The rate of screening for glucose-6-phosphate dehydrogenase deficiency in diabetes group was 3.6%,and the rate in the control group was 1. 1%.When HbAlc≥6.5%,the rate of screening for glucose-6-phosphate dehydrogenase deficiency in the diabetes group was signifi-cantly higher than that in the control group (χ2 =4.239,P =0.039).Conclusion The level of HbA1c is discordant with that of blood glucose in diabetic patients with glucose-6-phosphate dehydrogenase deficiency group.Diabetes leads to the increasement of the rate of glucose-6-phosphate dehydrogenase deficiency.The glucose-6-phosphate dehydrogenase activity of diabetic patients with non-glucose-6-phosphate dehydrogenase dificiency is higher than that of the normal group.It may be associated with the level of RBC or increase of compensatory.Further more,glucose-6-phosphate dehydrogenase activity may be a good indicator for controlling diabetes,which remains to be further studied.

Objective To investigate the association between the functional polymorphisms of Foxp3 gene and unexplained recurrent spontaneous abortion (URSA).Methods PCR-restriction fragment length polymorphism (rs3761548,rs2294021 ) and PCR with sequence-specific primers (rs2232365,rs5902434) were used to detect four polymorphisms of Foxp3 in 146 URSA cases and 112 normal controls.Results ( 1 ) The frequencies of rs3761548A/C were 10.3％,22.3％ in genotype C/C,38.4％,40.2％ in genotype A/C and 51.4％,37.5％ in genotype A/A between URSA patients and normal controls; the frequencies of rs2232365A/G were 5.5％,15.2％ in genotype A/A,47.9％,50.0％ in genotype A/G,46.6％,34.8％ in genotype G/G between URSA patients and normal controls; they all reached statistical difference ( P＜0.05 ).The carriers of rs3761548A allele and rs2232365G allele increased the risk of URSA (OR=1.73,1.61 ; all P ＜ 0.05 ).(2) There was no difference in the genotypic distribution of rs5902434del/ArTTpolymorphism between cases and controls ( P =0.10),but the frequency of del allele in URSA was statistically increased than that of controls (71.2％,62.5％ ; OR =1.49,P =0.04 ).(3) There was no different distribution in 3 genotypes (C/C,T/C,T/T) and 2 alleles (T and C) of rs2294021T/C between URSA patients and normal controls (P =0.18 and 0.08 ).(4) Estimated haplotype frequency distribution of rs5902434del/ATT,rs3761548A/C and rs22323565A/G showed haplotype del-A-G conferring the susceptibility to URSA ( OR =2.51,P ＜ 0.01 ) but haplotype del-C-G and ATT-A-A could provide protection on URSA ( OR =0.18,0.22 ; all P ＜ 0.01 ).Conclusion Functional polymorphisms of Foxp3 gene could probably confer the susceptibility to URSA,by altering Foxp3 function and (or) its expression.

By reverse transcription-polymerase chain reaction (RT-PCR), an open reading frame of pathogenesis-related protein 1 (PR1) was isolated from Panax notoginseng and named as PnPR1. Molecular and bioinformatic analyses of PnPR1 revealed that an open reading frame of 501 bp was predicted to encode a 166-amino acid protein with a deduced molecular mass of 18.1 kD. Homology analysis showed that the deduced amino acid sequence of PR1 protein of Panax notoginseng had a high similarity with other higher plants had the same conservative structure domain of cysteine-rich secretory protein (CAP). The recombinant expressed plasmid pET28a(+)-PnPR1 was expressed in Escherichia coli BL21. The expression conditions were optimized by induction at different times, different temperatures, different IPTG concentrations and different giving times. The optimum expression condition was 0.4 mmol.L-1 IPTG at 28 degrees C for 20 h. The successful expression of PnPR1 provides some basis for protein purification and preparation of the monoclonal antibody.

Objective To investigate the interference of hyperlipidemia and hyperbilirubinaemia to HbA1c measurements by ion‐exchange high‐performance liquid chromatography(IE‐HPLC) method .Methods Fresh whole‐blood samples collected with EDTA‐K2 anticoagulant tubes were divided into four groups :control group(HbA1c<6 .2% ) ,diabetes group(HbA1c≥6 .2% ) ,hyperlipi‐demia group(TG 3 -20 mmol/L);hyperbilirubinaemis group (TBIL 21 -549 μmol/L) .HbA1c of these samples were measured with affinity chromatography(AC‐HPLC) and IE‐HPLC respectively .Results When HbA1c≤18 .7% ,r=0 .993 ;95% confidence interval(CI) of HbA1c results by using IE‐HPLC method was -0 .71 -0 .89 ;coefficient of variation was -5 .8% -6 .8% ;P=0 .198 and the difference was not statistically significant .When HbA1c< 16 .3% ,r= 0 .997;95% CI of HbA1c results with IE‐HPLC method is -0 .31-0 .67;coefficient of variation was -5 .8% -4 .3% .P=0 .000 and the difference was statistically signifi‐cant .No interference was detectded with the results ;When HbA1c was 16 .3% -18 .7% ,positive bias was observed with the re‐sults .When TG≤20 .78 mmol/L ,r=0 .995;95% CI of HbA1c results with IE‐HPLC method was -0 .26-0 .50 ;coefficient of var‐iation was -5 .5% -5 .8% .P=0 .000 and the difference was statistically significant .No interference was detectded with the re‐sults;When TBIL≤549 .3 μmol/L ,r=0 .990 ;95% CI of HbA1c results with IE‐HPLC method was -0 .08 -0 .63;coefficient of variation was -14% -4 .1% .P=0 .000 and the difference was statistically significant .When TBIL≤342 .1 μmol/L ,r= 0 .994 ;95% CI of HbA1c results with IE‐HPLC method was -0 .09-0 .50;coefficient of variation was -5 .5% -4 .1% .No interference was detectded with the results .When TBIL was 380 .7-549 .3 μmol/L ,negative bias was observed with the results .Conclusion Our data indicated that HbA1c measurement with IE‐HPLC method could resist the interference of hyperlipidemia;When TBIL≤380 .7 μmol/L and HbA1c<16 .3% ,the results could meet the needs of general clinical detection .Clinical staff should choose more specific HbA1c measurement method according to the patient's condition .

OBJECTIVE: To track analysis of viral nucleic acid test results in patients with re-positive SARS-CoV-2 infection, and provide clinical reference for nucleic acid test of re-positive cases. METHODS: A retrospective study was conducted. The multiple nucleic acid results of 96 cases with SARS-CoV-2 infection tested by medical laboratory of Shenzhen Luohu Hospital Group from January to September in 2022 were analyzed. The test dates and cycle threshold (Ct) values of detectable positive virus nucleic acid in the 96 cases were summarized and analyzed. RESULTS: A total of 96 patients with SARS-CoV-2 infection were retested re-sampled for nucleic acid testing at least 12 days after the initial positive screening. Among them, 54 cases (56.25%) had Ct value of < 35 for nucleocapsid protein gene (N) and/or open reading frame 1ab gene (ORF 1ab), 42 cases (43.75%) had Ct value ≥ 35. In the re-sampling of infected patients, N gene titers were 25.08 to 39.98 Ct cycles, and ORF 1ab gene titers were 23.16 to 39.56 Ct cycles. Compared with the positive results of the initial screening, the Ct values of N gene and/or ORF 1ab gene positive were increased in 90 cases (93.75%). Among them, the patients with the longest duration of nucleic acid positive could still be positive for double targets (the Ct value of N gene was 38.60, and the Ct value of ORF 1ab gene was 38.11) at an interval of 178 days after the initial positive screening. CONCLUSIONS Patients infected with SARS-CoV-2 can be sustained or repeatedly tested positive for nucleic acid for a long period of time, and most of them had Ct values < 35. But whether it is infectious needs to be comprehensively evaluated by combining epidemiology, variant type, samples with the alive virus, and clinical symptoms and signs.
Humans ; COVID-19 ; SARS-CoV-2 ; Retrospective Studies ; Hospital Units ; Nucleic Acids