[Epidemiological characteristics of newly reported HIV-infected cases aged 50 years and above in Henan province, 1995-2020].

Objective: To analyze the epidemiological characteristics of newly reported HIV-infected cases aged 50 years and above in Henan province during 1995-2020, and to provide evidence for strategies on HIV/AIDS prevention and control. Methods: Information about newly reported HIV-infected cases aged 50 years and above in Henan between 1995 and 2020 were collected from the National Comprehensive HIV/AIDS Information System. The demographic and behavioral characteristics of HIV-infected cases aged ≥50 were analyzed, and the mixed linear model based on CD4+ T lymphocyte (CD4) counts back calculation was used to estimate the years, time and age of the HIV infection. Results: During 1995-2020, a total of 25 038 HIV-infected cases aged 50 years and above were newly reported, accounting for 25.8% (25 038/96 867) of the total number of newly reported HIV-infected cases in Henan. The proportion of newly reported cases over 50-years-old gradually increased from 4.5% (18/396) in 1995-2000 to 35.5% (9 666/27 239) in 2016-2020, with statistically significant difference (χ2=3 105.53, P<0.001). Among them, the proportion of HIV-infected cases aged 60 years and above were increasing year by year. The proportion of male cases were increasing along with ageing. The proportion of HIV-infected cases detected by medical institutions also showed an upward trend. The newly reported HIV-infected cases aged 50 years and above were mainly transmitted through sexual contact. The proportion of heterosexual transmission increased from 5.5% (1/18) in 1995-2000 to 86.2% (8 334/9 666) in 2016-2020, and the proportion of MSM-behavior-related transmission increased from 0.0% in 1995-2000 to 13.5% (1 304/9 666) in 2016-2020. The majority of cases had extra-marital and/or non-commercial heterosexual behavior (48.1%, 4 007/8 334) and the proportion showed an upward trend. The majority of male cases had commercial heterosexual behavior (54.9%, 3 169/5 775), and with increasing proportion along with the increase of age. The majority of female cases had extra-marital and/or non-commercial heterosexual behavior (62.5%, 1 600/2 559), with increasing proportion of extra-marital and/or non-commercial heterosexual behavior. The proportion of heterosexual behavior with spouse or stable sexual partners showed a downward trend. The estimations based on CD4 counts back calculation model showed that among the newly reported HIV-infected cases aged 50 years and above, the average age being infected was (54.8±10.2) years, with 33.8% (4 263/12 621) infected before 50. The interval between infection and diagnosis was (5.7±6.2) years, of which 52.6% (6 636/12 621) were infected for 5 years or longer and 34.7% (4 384/12 621) were in the last 3 years. There was no linear correlation trend in the composition of infection years among the newly reported HIV-infected cases over 50-years-old. Conclusions: In Henan, from 1995 to 2020, the number of newly reported HIV-infected cases aged 50 years and above was increasing and sexual transmission becoming the main transmission route. The increase of prevalence was mostly seen in 60-years-old men, low education level and detected mainly by medical institutions. For this age group, the focus of HIV/AIDS prevention and control should target on those who were transmitted through extra-marital and/or non-commercial heterosexual, commercial heterosexual and MSM behavior and it is necessary to strengthen the HIV testing and detection in this population and in the elderly floating group.

[Research progress in radiomics based on CT and MRI images for prediction of microvascular invasion in hepatocellular carcinoma].

Hepatocellular carcinoma (HCC) is one of the most common malignant tumors worldwide. Microvascular invasion (MVI) is considered the major risk factor for postoperative recurrence and metastasis in HCC. The diagnosis of MVI relies on the postoperative pathological assessment of the tumor tissues. Seeking non-invasive methods and biomarkers for evaluation of MVI before surgery has important clinical implications for guiding surgical treatment and improving patients' survival. Recent studies have reported the applications of radiomics technique in prediction of MVI in HCC and showed promising results. Herein we summarized the research progress in CT- or MRI-based radiomics models for prediction of MVI in HCC to provide helpful thinking for further research in this field.

[Knockdown of mediator complex subunit 19 suppresses the migration and invasion and epithelial-mesenchymal transition in prostate cancer PC3 cells].

Objective: To studythe effect of lentivirus-mediated inhibition of Med19 on cell migration andinvasion in the PC3 cells, and explore the mechanism of epithelial-mesenchymal transition transformation. Methods: The lentivirus vectors containing small interferingRNA(siRNA) targeting Medl9 gene were constructed and transfected to PC3 cells.Quantitative real-time PCR(qRT-PCR) and Western blot analysis were used to detect the Medl9 expression in the siRNA group(PC3-Med 19-siRNA cells)and the NC group(PC3-Med 19-sc cells) at 72h after the transfection.The cell mobility,migration and invasion ability of PC3 cells were respectively measured by Boyden migration and woun-healing assay. The expression of E-Cadherin, N-Cadherin, Vimentin, ZEB2, Snail-1, and Snail-2 mRNA were detected by using qRT-PCR. Results: The expression of Medl9 mRNA in PC3-Med 19-siRNA cells was lower than that in PC3-Med 19-scRNA cells(P<0.01). The number of migrated cells and invaded cells were significantly decreased in PC3-Med 19-siRNA cells(P<0.01). The expression of N-Cadherin, Vimentin, ZEB2, Snail-1, and Snail-2 mRNA were remarkablylower and E-Cadherin was higher in PC3-Med 19-siRNA cells. Conclusion: Med 19 inhibitioncouldreduce migration abilityof prostate cancer PC3 cells by epithelial-mesenchymal transition.

[Identification and evaluation on methods with upstream flank sequences of CRISPR1, regarding Escherichia coli and Shigella].

Objective: To analyze the effect of the identification and evaluation of Escherichia (E.) coli and Shigella, based on the upstream flanking sequences of CRISPR1. Methods: Both CRISPR and cas sequences were obtained through the BLAST with repeating sequences against the publicly complete genome in GenBank that related to E. coli and Shigella. Clustal X was used to perform multi-sequences alignment of the flanking sequences. PCR method was used to amplify the upstream flanking sequences of CRISPR1 in order to appraise the effect of identification and evaluation of upstream flanking sequences on E. coli and Shigella, which were based on the upstream flanking sequences of CRISPR1. Results: The results showed that 73.4% of the strains containing the I-E CRISPR/Cas that belonged to the phylogroups A, B1, D while 8.4% strains carried the I-F CRISPR/Cas. Another 17.2% of the strains owned CRISPR3-4 (non-CRISPR/Cas) only belonged to the phylogroups B2. All the Shigella strains carried I-E CRISPR/Cas. More than 99% of similarity the CRISPR1 upstream-flanking sequences was seen in E. coli (except B2) and Shigella and E. coli (B2). Both sensitivity and specificity were greater than 91% after PCR amplification in the region to identify the E.coli and Shigella. Conclusion: The upstream of CRISPR1 could achieve a preliminary identification effect on E.coli and Shigella.

[Association between phage-mediated shiga toxin and molecular distribution of CRISPR in Escherichia coli O26 ∶ H11 or NM].

Objective: To investigate the association between phage-mediated shiga toxin and molecular distribution of CRISPR in Escherichia (E.) coli O26∶H11 or NM. Methods: A total of 135 E. coli O26 ∶ H11 or NM strains were collected from NCBI database. Software CRT and CRISPR Finder were used to extract CRISPR and Excel was used to assign the spacer of unique number and type CRISPR. And the relationship between CRISPR and stx phage was analyzed. Results: All the 135 E. coli O26 ∶ H11 or NM strains had the CRISPR. For CRISPR1, CRISPR2.1, CRISPR2.2 and CRISPR3-4, 19, 22, 1 and 1 subtypes were found, respectively. According to the four CRISPR sites, the strains could be divided into 40 subtypes. Stx-phage was only observed in the group C of CRISPR. Compared with E. coli of stx-phage negative, E. coli with stx-phage harbored more spacers. Conclusions: CRISPR loci was extensively existed in E. coli O26∶H11 or NM, and many subtypes were found in these strains. The presence of stx-phage was related to the molecular distribution of CRISPR in E. coli O26∶H11 or NM. CRISPR might be a valuable biomarker to identify strains with high virulent potential.

Comparison of computed tomography and magnetic resonance imaging in the evaluation of facet tropism and facet arthrosis in degenerative cervical spondylolisthesis.

The aim of this study was to determine the reliability of magnetic resonance imaging (MRI) in the assessment of facet tropism and facet arthrosis of spondylolisthesis levels in degenerative cervical spondylolisthesis as compared to computed tomography (CT). The discrepancies in the interpretation of CT and MRI data in the evaluation of facet tropism and arthrosis have given rise to questions regarding the reliability of comparisons of the two techniques. Using a 4-point scale, 3 blinded readers independently graded the severity of facet tropism and facet arthrosis of 79 cervical facet joints on axial T2-weighted and sagittal T1 and T2-weighted turbo spin echo images as well as the corresponding axial CT scans. All results were subjected to the kappa coefficient statistic for strength of agreement. In the assessment of the severity of facet arthrosis, intermethod agreement (weighted κ) between CT scanning with a moderate inter-rater reliability (range κ = 0.43-0.57) and MRI with fair inter-rater reliability (range κ = 0.23-0.38) was 0.76 and 0.43 for the severity of facet tropism and facet arthrosis, respectively. Intra-rater reliability for the severity of facet arthrosis was moderate to substantial for CT and was moderate for MRI scans. Intra-rater reliability for the severity of facet tropism was substantial to very good for CT and substantial for MRI scans. MRI can reliably determine the presence or degree of facet tropism but not facet arthrosis. Therefore, for a comprehensive assessment of cervical facet joint degeneration, both a CT and an MRI scan should be performed.

Two adult galactosaemia females with normal ovarian function and identical GALT mutations (Q188R/R333G).

We report two unrelated cases of adult galactosaemia females with normal ovarian function and Q188R/R333G mutations. Clinical history has been followed for 40 years. Biochemical finding in one patient are consistent with the presence of small amounts of galactose-1-phosphate uridyltransferase (GALT) activity, which differs from classical galactosaemia.

Antistress effect of oligosaccharides extracted from Morinda officinalis in mice and rats.

AIM: To explore the antistress effect of Morinda officinalis (Chinese medicine "Bajitian") oligosaccharides (MW-97) in mice. METHODS: Male mice and rats were subjected to a variety of unpredictable stressors on a daily basis over 15 d period and then the Vidiomex-V image pattern analytic system was used to observe the spontaneous motor activity. Meanwhile, regular method was used to prepare pathologic section of adrenal gland and blood cells analytic system was used to detect white blood cell (WBC) count (including relative WBC percentages) in peripheral blood. On the end, the serum level of stress hormone was detected using radioimmunoassay. RESULTS: Chronic stress resulted in diffuse hyperplasia of the adrenal cortex and atrophy of the adrenal medulla in mice, which suggested that stress-adaption failure of the adrenal gland occurred, while adrenal gland of the mice pretreated with MW-97 (100 mg/kg, ip) prior to each stressor for 15 d did not occur any pathologic changes. In addition, chronic stress also significantly reduced the WBC count and relative WBC percentages in the peripheral blood, including the percentage of lymphocytes, monocytes, neutrophils, however, MW-97 (25 and 100 mg/kg) reversed these changes and raised WBC count, along with relative WBC percentages significantly. Furthermore, the serum concentration of testosterone was decreased and corticosterone was increased significantly in chronically stressed animals. MW-97 also declined the serum level of corticosterone and raised level of testosterone. MW-97 had no effects on the spontaneous motor activity in the stressed mice. CONCLUSION: MW-97 had antistress effect against chronic stress, moreover, MW-97 had no excitatory or inhibitory effects on the CNS, which suggested that MW-97 might become a new kind of antistress agent.

Surgical treatment of nonunited fractures of the odontoid process, with special reference to occipitocervical fusion for unreducible atlantoaxial subluxation or instability.

Fifty-seven consecutive patients treated surgically for nonunited fractures of the odontoid process were reviewed. All patients presented late, exhibiting neurological deficits subsequent to nonunion. Delay in presentation was between 6 and 120 months (mean 32 months) after the original injury, due to missed diagnosis or inappropriate management. Seven patients who were reduced in traction underwent a Gallie atlantoaxial fusion. In the remaining 50 patients who were unreducible, an occipitocervical arthrodesis was performed. They were followed up for a minimum of 2 years, except one who died from postoperative respiratory failure. All patients obtained a solid bony union, including two in whom nonunion occurred following atlantoaxial fusion, and occipitocervical fusion was added as a rescue. Thirty-eight patients achieved excellent neurological recovery, nine still had some disability, five retained their neurological deficits and two reported a deterioration. In two patients, a recurrence in a traumatic episode was experienced long after a resolution. Our findings demonstrate that occipitocervical arthrodesis is preferable for unreducible subluxation or instability of atlantoaxial articulation in nonunion of odontoid fractures.

Effect of mobile phase composition on the separation of propranolol enantiomers using a perphenylcarbamate beta-cyclodextrin bonded chiral stationary phase.

An analytical column packed with a novel perphenylcarbamate beta-cyclodextrin bonded chiral stationary phase was used to separate propranolol enantiomers. Good separation results were obtained using triethylammonium acetate (TEAA) buffer and methanol mixtures as the mobile phase. Effects of the methanol concentration, buffer concentration and pH value on the retention and the enantioselectivity of propranolol enantiomers were investigated on this column. The retention times and the separation factor decrease with increase of the methanol concentration as expected in reversed-phase HPLC. At trace TEAA amount, the solute eluted out with anti-Langmuirian band profiles, their retention times decreased quickly with increase of TEAA concentration and attained a minimum at a TEAA concentration of 20 ppm. Above 20 ppm, solute band profiles changed to a Langmuirian shape, the retention times of enantiomers increased with increasing buffer concentration, and eventually, they attained asymptotes at ca. 1% TEAA. A simulation considering the different interactions between the solute and the additive at above and below 20 ppm TEAA concentration as well as system peaks interference can successfully explain the anti-Langmuirian band profiles and the retention time variation trend.

Molecular and biochemical basis of galactosemia.

Galactosemia is a clinically heterogeneous autosomal recessive inborn error of metabolism caused by deficiency of galactose-1-phosphate uridylyltransferase (GALT). Despite the numerous point mutations identified in the GALT gene, the prevalence of these mutations in different ethnic groups has not been studied. Reports on genotype/phenotype correlation are not consistent due to the small sample sizes studied and the lack of a sensitive enzyme assay. We applied multiplex PCR/ASO dot blot analysis to screen 293 galactosemic patients for 17 known point mutations in exons 5, 6, and 10. Our data demonstrate that only 7 of these mutations were detected in our patients, accounting for 65% of the GALT mutant alleles. Although Q188R is the most common mutation in Caucasian and Hispanic patients, the S135L mutation is most common in African-Americans. Another mutation, F171S, was observed only among African-American patients. An improved, sensitive, and accurate method was used to measure GALT activity in patient's red blood cells. The results indicated that patients homozygous for Q188R have no enzyme activity while those homozygous for S135L had residual enzyme activity. Interestingly, both Q188R/S135L and S135L/F171S compound heterozygotes demonstrated zero enzyme activity. Overall, 85% of Q188R compound heterozygotes also did not have any enzyme activity, whereas the remaining Q188R and the majority of S135L compound heterozygotes expressed variable amounts of GALT activity. We speculate that heterodimeric subunit interaction plays an important role in determining the overall enzymatic activity. Various genotypes thus result in biochemical and clinical heterogeneity among the patients.

Radiochemical assay of minute quantities of galactose-1-phosphate uridyltransferase activity in erythrocytes and leukocytes of galactosemia patients.

A sensitive radioisotopic method has been developed which can detect galactose-1-phosphate uridyltransferase (GALT) activity as low as 0.1% of normal control values in both erythrocytes and leukocytes. This assay utilizes carbon-14 labeled galactose-1-phosphate with high specific activity and requires removal of endogenous galactose-1-phosphate (Gal-1-P) and uridine diphosphate glucose (UDPGlc) through dialysis. Optimal exogenous UDPGlc concentration has been determined with a fixed concentration of Gal-1-P in the incubation. The rate of product, uridine diphosphate galactose (UDPGal), formation is monitored at three different times. Among 423 patients with galactosemia studied by this method, 363 patients exhibited no detectable GALT activity in their erythrocytes and 60 patients were found to have detectable erythrocyte GALT activity ranging from 0.02 to 5.0 units normal values: > 20 units). The former group of patients was designated as classic galactosemia (GG) and the latter group as galactosemia variant (GV). Leucocytes from ten patients belonging to the GG group also showed complete absence of GALT activity while leukocytes from two patients belonging to the GV group showed GALT activity at levels comparable with those found in their erythrocytes. Because there is extensive biochemical heterogeneity among galactosemia patients, we recommend that an assay with increase sensitivity be carried out on blood samples from galactosemia patients so that clinical, biochemical and molecular correlations made by different groups of investigators can be compared.

Correlative study on findings of dynamic myelography and surgical operation in non-bony lumbar spinal canal stenosis.

The authors performed dynamic lumbar myelography with Omnipaque on 110 patients from 1990 to 1992, of them, 33 cases were diagnosed as non-bony lumbar spinal canal stenosis according to contrast medium defect seen on the lateral view of the myelogram. All such cases were operated on and their dynamic pathological findings during the operation were recorded to compare with the abnormal findings observed on the myelograms, there was a high coincidence rate of 88.7%. The authors believe that the fibrous degenerative changes make up the basis of dural sac compression and the dynamic compression in the spinal canal plays a very important role in causing the severity of the stenosis. As the dynamic pathological findings on the myelograms can not be well demonstrated on CT scanning and MR imaging, the dynamic lumbar myelography should be the method of choice for use in some cases.

Biochemical and molecular studies of 132 patients with galactosemia.

We evaluated 132 galactosemia patients for the Q188R (glutamine-188 to arginine) mutation in the human galactose-1-phosphate uridyltransferase (GALT) gene and for GALT activity in their hemolysates by a sensitive radioisotopic method. In those without any detectable GALT activity (GG), the Q188R mutation constituted 67% of the alleles. In patients with detectable GALT activity (GV), only 16% of the alleles were accounted for by Q188R. In all patients who were homozygous for the Q188R mutation, no erythrocyte GALT activity could be demonstrated. There was an extensive variation in the amount of detectable GALT activity ranging from 0.1% to 5% of the normal values among the GV patients. There was a difference in the frequency of Q188R mutation in the GALT alleles among patients belonging to different racial and ethnic groups. In Caucasian and Hispanic patients, the frequency was not far different (64% and 58%, respectively). On the other hand, only 12% of the GALT alleles with Q188R were found in African-American patients.

Correlation of cognitive, neurologic, and ovarian outcome with the Q188R mutation of the galactose-1-phosphate uridyltransferase gene.

This study was conducted to determine whether there is a genotype/phenotype correlation between aspects of cognitive, neurologic, and ovarian outcome in patients with galactosemia and the Q188R mutation of the galactose-1-phosphate uridyltransferase gene. The results showed that the Q188R mutation was found in 72% of alleles: 38 patients were homozygous and 21 were heterozygous for Q188R; eight patients did not have the mutation. The mean Broad Cognitive score for the group homozygous for Q188R was 75 (SD = 16), which was not statistically different from the outcome for the heterozygous group (mean score, 67; SD = 25) or the negative group (mean score, 88; SD = 21). Tremor, ataxia, and dysmetria were found in 12 subjects, and there was no association with Q188R status. Similarly, there was no association of this mutation with the development of primary amenorrhea (8 subjects) versus secondary amenorrhea (found in 14 women). Our findings suggests that the variability of outcome for patients with classic galactosemia cannot be explained by Q188R status alone, at least with regard to cognitive functioning, presence of neurologic symptoms, and timing of the onset of ovarian failure.

Kinetics of irreversible inhibition of creatine kinase during modification by o-phthaldehyde.

It has been previously reported that, with a fluorescence probe formed from o-phthaldehyde (OPTA) and the thiol and amino groups at or near the active site of creatine kinase, inactivation and exposure of the probe take place simultaneously and well before unfolding of the molecule as a whole. In this study, the inactivation and modification kinetics of purified rabbit muscle creatine kinase by OPTA have been compared, the former by following the substrate reaction in the presence of a previously described inactivator. The microscopic rate constants for the reaction of the inactivator with the free enzyme and with the enzyme-substrate complexes were determined. From the results obtained it appears that OPTA is noncompetitive with respect to both substrates. The inactivation kinetics is monophasic with OPTA, and neither ATP nor creatine alone affect the rate constant of inactivation of the enzyme, indicating that the irreversible inhibition of creatine kinase by OPTA is of the noncompetitive type.