Whole-genome sequencing of bacteria has recently emerged as a cost-effective and convenient approach for resolving many microbiological questions.Here, the current status of whole-genome sequencing in clinical microbiology and the main problems that exist were reviewed, focusing on six essential tasks: identifying and culturing the species of an isolate, rapid identification of pathogen properties, monitoring the emergence and spread of an epidemic outbreak, developing vaccine and monitoring its variation,metagenomics and single-cell sequencing.The authors predicted that the application of whole-genome sequencing will soon be sufficiently fast, accurate and cheap to be used in routine etiological detection, even though there is still a long way to go.

Background Trachoma is a serious blinding eye disease.At present,the incidence rate of trachoma in China has been greatly reduced with the unremitting efforts of World Health Orgnization (WHO) and Chinese government.However,in some economically underdeveloped areas,there is still active trachoma.Ohjective This study aimed to survey the prevalence of children's active trachoma in rural area of Naxi district and Jiangyang district of Luzhou city and assess the prevention and treatment outcome of trachoma during 10 years.Methods A cross-sectional study was performed in Naxi district and Jiangyang district of Luzhou city,the economic underdeveloping and water-starved regions in the year 2000.The active trachoma was screened in 214 school-age children with the male 113 and female 101 based on simplified trachoma grading system and trachoma rapid assessment survey standards and requirements of WHO.The symptoms of active trachoma were divided into trachomatous follicular (TF),trachomatous inflammation intense (TI),trachomatous scarring (TS),trachoma trichiasis (TT) and corneal opacities (CO).The epidemiological survey was re-assessed in 100 school-age children (with the male 58 and female 42) in these regions in the year 2013 to evaluate the prevalence change of children active trachoma during 10 years.Results In the year 2000,214 children were screened in 4 villages of Naxi district and Jiangyang district,and the prevalence rate of TF was 49.07％ (105/214) in the year 2000.The prevalence rate of TF was 49.05％ (52/106) in the Naxi district and 49.07％ (53/108) in the Jiangyang district,showing no significant difference between them (x2 =0.01,P =0.99).In the 100 school-age children who received survey in Naxi district and Jiangyang district in the year 2013,no TF,TF,TI,TS,TT and CO was found,showing no active trachoma.Conclusions The active trachoma has been controlled in rural areas of Luzhou city following the prevention and treatment of trachoma and the improvement of economic status in past decade.

In recent years , with the increasing of immunocompromised patients , Trichosporon spp. has become a more and more significant clinical opportunistic pathogen .Medical staff should enhance the clinical awareness to this pathogen . In this paper , the classification , virulence factor and pathogenic mechanism, infections, diagnostic methods of laboratory , antifungal susceptibility and treatment of Trichosporon spp. were reviewed systematically . Trichosporon asahii is the major pathogen of invasive infections.Biofilm formation and enzyme production will promote its ability to escape from antifungal drugs and host immune responses .Matrix assisted laser desorption ionization time of flight mass spectrometry has the advantages of accurate , fast and low cost for identification of Trichosporon spp., and zoles is the first-line treatment for invasive infections .

Objective To investigate the in vitro antibacterial activity of single cefoperazone,ceftazidime,imipenem,tigecycline and colistin and their combination with sulbactam against clinical isolates of Acinetobacter baumannii (A.baumannii).Methods Twentythree meropenem-resistant A.baumannii strains and 21 meropenem-sensitive strains from the Study for Monitoring Antimicrobial Resistance Trends (SMART) during 2011 and 2012 were collected,and their combination susceptibility tests were performed by the checkerboard dilution method.The fractional inhibitory concentration (FIC) index was calculated to evaluate the combination effect of antibiotics.Results No antagonism effect was observed for all of combinations.The combination of sulbactam with cefoperazone or tigecycline mainly produced synergistic effect on A.baumannii,and the strains with FIC index ≤0.5 accounted for 56.8％ and 50.0％,respectively.The combination of sulbactam with imipenem or colistin showed synergistic or partially synergistic effect on A.baumannii,and the strains with FIC index ＜ 1 accounted for 61.4％ and 52.3％,respectively.However,the combination of sulbactan with ceftazidime didn't show any interation,and the strains with FIC index ≥4 accounted for 63.6％.Conclusion The combination of sulbactam with cefoperazone has the best synergistic effect on A.baumannii,especially on carbapenem-sensitive A.baumannii.The combination of sulbactam with imipenem or tigecycline may enhance the antibacterial activity on carbapenem-resistant A.baumannii.The combination of sulbactam with imipenem or tigecycline may be helpful for the treatment of carbapenem-resistant A.baumannii infection.

Objective: To estimate the transmission capacity of influenza clustering in schools and nurseries, and to evaluate the effect of suspension measures, providing a basis for formulating disease management strategies and control measures. Methods: The SEIAR dynamics model was used to simulate the epidemic data, calculating the basic regeneration coefficient R 0 of the epidemic to evaluate the epidemic transmission capacity, and calculating the cumulative incidence rate of the epidemic to evaluate the prevention and control effect of the suspension measures. Results: The basic regeneration coefficient R 0 was 8.44(8.01,8.89) without intervention. There were statistically significant differences in R 0 of influenza epidemic among different types of school(F=9.52, P<0.01). The R 0 of influenza epidemic in primary and secondary schools were higher than that in nurseries(P<0.05). R 0 of influenza A was higher than that of influenza B(t=2.71, P<0.01). R 0 of influenza A(H3) was higher than of influenza B(Victoria)(P<0.05). The cumulative incidence of the outbreaks which were suspended for 4 days and 7 days was significantly lower than that in the non-suspensions(P<0.05). However, there was no significant difference in the cumulative incidence of the outbreaks between the 4-day suspension and the 7-day suspension(P>0.05). Conclusion Transmission capacity of school-based influenza epidemic is high, especially among primary and secondary schools. When the epidemic situation of infected class meets the suspension standard, it is recommended to suspend classes for 4 days.

Objective To explore the expression and significance of miR-21 in patients with primary gout. Methods The patients were divided into 4 groups: 35 acute gout patients (AG), 50 intermittent gout patients (IG), 25 chronic gout patients (CG) and 39 healthy patients. Their peripheral blood were collected and laboratory indexes were recorded. The expression of miR-21 and Nod-like receptor pyrin domain-containing protein 3 (NLRP3) mRNA in the peripheral blood mononuclear cells (PBMCs) was detected by real-time quantitative polymerase chain reaction (RT-qPCR). The blood and clinical data of another 5 healthy volunteers were collected, their peripheral blood was stimulated with 100 μg/ml monosodium urate (MSU) for 1 hour, pho-sphate buffer (PBS) was used as controls, then the expression of microRNA (miR)-21, NLRP3, interleukin (IL)-1β mRNA was detected by RT-qPCR. Rank sum test and spearman correlation analysis were used for data analysis. Results In primary gout patients, the expression of miR-21 in AG [12 ×10-4 (8.0 ×10-4)], IG [9.4 ×10-4 (6.9 ×10-4)], CG [7.3 ×10-4 (5.6 ×10-4)] was significantly higher than that in healthy control group [1.0×10-4(2.0×10-4)] (Z=9.83, P=0.02], while the expression of NLRP3 in AG[0.0444(0.0233)], IG[0.0581(0.0326)], CG[0.0314(0.0198)] was significantly lower than that in healthy control group [0.0886(0.0359)] (Z=13.82, P<0.01). In the primary gout of IG group, the expression of miR-21 was positively correlated with NLRP3 mRNA (r=0.449, P=0.016). After stimulated by 100 μg/ml MSU, the expression of miR-21 of the stimulated group [8.78×10-4(14×10-4)] was higher than that in the control group [6.25×10-4(6×10-4)](Z=-2.203, P<0.05), and the expression of IL-1βin stimulated group [3.06(2.00)] was higher than that in the control group [2.64 (1.22] (Z=-2.203, P<0.05). The level of miR-21 in patients with primary gout was positively correlated with the level of uric acid (UA), glutamic oxaloacetic transaminase (AST) and glutamic pyruvic transaminase (ALT) (r=0.473, 0.639, 0.487, P<0.05). Conclusion The increase of miR-21 in patients with primary gout may be involved in the inflammatory reaction of gout.

Objective:To investigate the combined effect of fluoride exposure and low nutrition on osteogenesis and osteoclastic differentiation in rats.Methods:SD rats were divided into four groups by the method of random number table, namely normal nutrition group, low nutrition treatment group, fluoride exposure group and co-treatment of fluoride and low nutrition group according to 2 × 2 factorial experimental design, 8 rats in each group, half male and half female. Five months after the experiment, immunohistochemistry was used to test the expression levels of femoral alkaline phosphatase (ALP), runt-related transcription factor 2 (Runx2), osteoprotegerin (OPG) and receptor activator of nuclear factor kappa B ligand (RANKL). Analysis of variance of factorial design was used to determine the interaction between fluoride exposure and low nutrition on osteogenesis and osteoclastic differentiation.Results:The immunohistochemical results of bone tissue showed that there were significant differences in the expression levels of osteogenesis differentiation markers ALP and Runx2 between different groups ( F = 25.98, 17.77, P < 0.001). Compared with normal nutrition group (0.005 2 ± 0.002 7, 0.003 1 ± 0.001 4), the expression levels of ALP and Runx2 in fluoride exposure group were higher (0.019 5 ± 0.005 0, 0.014 4 ± 0.004 4, P < 0.05). There was no significant difference between low nutrition treatment group (0.002 6 ± 0.001 8, 0.004 4 ± 0.003 2) and co-treatment of fluoride and low nutrition group (0.003 6 ± 0.000 7, 0.002 9 ± 0.000 8, P > 0.05). The expression levels of ALP and Runx2 in co-treatment of fluoride and low nutrition group were lower than those of fluoride exposure group ( P < 0.05). There were significant differences in the expression level osteoclastic differentiation marker of RANKL and the ratio of RANKL/OPG ( F = 10.50, 31.05, P < 0.001). Among them, the RANKL/OPG ratio (0.115 3 ± 0.039 5) in fluoride exposure group was lower than that in normal nutrition group (1.426 3 ± 0.777 2), and the RANKL expression level and RANKL/OPG ratio (0.019 5 ± 0.007 7, 7.258 7 ± 3.674 3) in co-treatment of fluoride and low nutrition group were higher than those in normal nutrition group (0.004 4 ± 0.002 5, 1.426 3 ± 0.777 2, P < 0.05). However, there was no significant difference in the RANKL expression level and RANKL/OPG ratio (0.004 0 ± 0.001 9, 2.022 3 ± 0.753 7) in low nutrition treatment group ( P > 0.05). The expression level of RANKL and the ratio of RANKL/OPG in the co-treatment of fluoride and low nutrition group were higher than those in low nutrition treatment group and fluoride exposure group ( P < 0.05). The 2 × 2 analysis of variance of factorial design showed that fluoride exposure and low nutrition had interaction on ALP, Runx2, RANKL expression levels and RANKL/OPG ratio ( F = 4.38, 19.39, 22.12, 108.00, P < 0.05), antagonistic effect on ALP and Runx2 expression, synergistic effect on RANKL expression and RANKL/OPG ratio. Conclusions:In rat bone tissue, fluoride exposure promotes osteogenesis differentiation, inhibits osteoclastic differentiation dominated by active osteogenic function. The interaction between fluoride and low nutrition on osteogenesis and osteoclastic differentiation is antagonistic osteogenesis differentiation and synergistic promotion of osteoclastic differentiation. Normal nutrition conditions are material basis of osteogenesis differentiation, and low nutrition is the inducement of enhanced osteoclastic differentiation.

Objective:To learn about the levels of 5-methylcytosine (5-mC) and 5-hydroxymethylcytosine (5-hmC) in bone tissue of rats with different types of skeletal fluorosis and analyze their correlation.Methods:Thirty 4-week-old SPF grade healthy SD rats were selected. After adaptive feeding for 1 week, the rats were divided into control group (4 ml·kg -1·bw deionized water + standard maintenance diet), osteosclerosis group [20 mg·kg -1·bw sodium fluoride (NaF) + standard maintenance diet], and osteoporosis/osteomalacia group (20 mg·kg -1·bw NaF + low-calcium and low-protein partial diet) according to their body weight (100 - 120 g) by random number table method, with 10 rats in each group, half male and half female; gavaged 6 days each week and the experimental period was 5 months. At the end of the experiment, samples of rat heart blood and lower limb femur were collected. The contents of serum methyl donor S-adenosylmethionine (SAM) and its metabolite S-adenosylhomocysteine (SAH) in serum, and the levels of 5-mC and 5-hmC in bone tissue were measured by enzyme-linked immunosorbent assay (ELISA). Western blot was used to determine the expression of DNA methyltransferase (DNMTs) and DNA hydroxymethylase (TETs) in bone tissue of rats. The correlation between serum SAM content, SAM/SAH ratio and bone tissue 5-mC level, and between the bone tissue 5-mC level and 5-hmC level was analyzed. Results:Serum SAM [11.03 (7.06, 18.63), 3.96 (2.32, 9.09), 3.91 (2.35, 4.46) nmol/L], SAH content [(4.69 ± 0.55), (5.41 ± 1.13), (13.90 ± 1.09) ng/L], SAM/SAH ratio [2.58 (1.54, 4.12), 0.62 (0.52, 1.69), 0.14 (0.13, 0.15)] and bone tissue 5-mC [103.39 (97.37, 109.35), 52.50 (50.19, 68.13), 55.03 (49.97, 59.57) ng/L], 5-hmC levels [(32.61 ± 8.84), (56.96 ± 8.48), (20.34 ± 6.22) ng/L] in the control group, osteosclerosis group and osteoporosis/osteomalacia group were compared, and the differences were statistically significant beween three groups ( H/ F = 12.81, 284.24, 21.85, 19.37, 55.23, P < 0.01). Compared with the control group, the content of SAM, the ratio of SAM/SAH, the level of 5-mC in the osteosclerosis group and osteoporosis/osteomalacia group, and the level of 5-hmC in the osteoporosis/osteomalacia group were lower ( P < 0.05), while the content of SAH in the osteoporosis/osteomalacia group and the level of 5-hmC in the osteosclerosis group were higher ( P < 0.05). Compared with the osteosclerosis group, the content of SAH in the osteoporosis/osteomalacia group was higher, while the ratio of SAM/SAH and the level of 5-hmC were lower ( P < 0.05). Western blot showed that there were statistically significant differences in the expression levels of DNMT1, DNMT3A, DNMT3B, TET1 and TET2 protein in bone tissue of rats in the control group, osteosclerosis group, and osteoporosis/osteomalacia group ( F = 285.45, 345.58, 239.83, 311.52, 318.24, P < 0.001). Among them, the expression levels of DNMT1, DNMT3A and DNMT3B protein in the osteosclerosis group and osteoporosis/osteomalacia group were lower than those in the control group, and the expression levels of DNMT1, DNMT3A and DNMT3B protein in the osteosclerosis/osteomalacia group were lower than those in the osteosclerosis group ( P < 0.05); the expression levels of TET1 and TET2 protein in osteosclerosis group were higher than those in the control group and osteoporosis/osteomalacia group, and the expression levels of TET1 and TET2 protein in the osteoporosis/osteomalacia group were lower than those in the control group ( P < 0.05). The results of Spearman rank correlation analysis showed that the content of SAM and the ratio of SAM/SAH in the control group, osteosclerosis group and osteoporosis/osteomalacia group were positively correlated with the level of 5-mC in bone tissue ( rs = 0.89, 0.92, 0.81, 0.73, 0.87, 0.73, P < 0.05). The levels of 5-mC and 5-hmC in bone tissue of rats in each group were negatively correlated ( rs = - 0.69, - 0.68, - 0.72, P < 0.05). Conclusions:The level of 5-mC in bone tissue of osteosclerotic fluorosis rats is low, and the level of 5-hmC is high, while those of osteoporosis/osteomalacia fluorosis rats are lower. The difference of 5-mC level in bone tissue of rats with different types of skeletal fluorosis is not significant, which may be related to the difference of 5-hmC level in bone tissue.

AIM: To evaluate clinical efficacy and safety of ultrasound cycloplasty(UCP)in the treatment of refractory glaucoma.METHODS:From June 2021 to October 2022, a total of 17 patients(17 eyes)with refractory glaucoma were enrolled in this prospective study, and they all underwent UCP. The patients underwent 6 mo followed-up post-treatment, and the intraocular pressure(IOP), pain grade score, IOP lowering drugs, success rate and occurrence of complications were documented.RESULTS:The IOP was significantly decreased from 51.98±7.80 mmHg before UCP to 32.54±13.21 mmHg at 1 d, 22.38±11.98 mmHg at 1 wk, 22.63±10.78 mmHg at 1 mo, 26.05±9.17 mmHg at 3 mo, and 23.73±9.60 mmHg at 6 mo postoperatively(all P<0.01). The percentage of IOP reduction after treatment was 36.25%, 57.10%, 56.35%, 49.16% and 54.09% at 1 d, 1 wk, 1, 3, and 6 mo, respectively. The pain grade scores were decreased(P<0.01). There was a statistically significant reduction in the use of IOP lowering medications(P=0.008). At 6 mo postoperatively, 2 eyes(12%)were complete success, 11 eyes(65%)were qualified success, and 4 eyes(24%)were failure. The main complication observed was anterior chamber inflammation in 1 eye(6%), foreign body sensation in 2 eyes(12%), subconjunctival hemorrhage in 2 eyes(12%), and conjunctival congestion in 6 eyes(35%). All symptoms spontaneously resolved within 1 wk without requiring any specific treatment. One case of choroidal detachment(6%)occurred on 10 d postoperatively, but recovered after orally treated by prednisone acetate for 1 mo. No other serious complications, such as hyphema, corectopia, synechia or macular edema were reported.CONCLUSION:UCP is safe and efficacious in treating refractory glaucoma, reducing IOP and alleviating ocular pain symptoms, while maintaining a favorable safety profile.