A novel amphiphilic motif at the C-terminus of FtsZ1 facilitates chloroplast division.

In bacteria and chloroplasts, the GTPase filamentous temperature-sensitive Z (FtsZ) is essential for division and polymerizes to form rings that mark the division site. Plants contain two FtsZ subfamilies (FtsZ1 and FtsZ2) with different assembly dynamics. FtsZ1 lacks the C-terminal domain of a typical FtsZ protein. Here, we show that the conserved short motif FtsZ1Carboxyl-terminus (Z1C) (consisting of the amino acids RRLFF) with weak membrane-binding activity is present at the C-terminus of FtsZ1 in angiosperms. For a polymer-forming protein such as FtsZ, this activity is strong enough for membrane tethering. Arabidopsis thaliana plants with mutated Z1C motifs contained heterogeneously sized chloroplasts and parallel FtsZ rings or long FtsZ filaments, suggesting that the Z1C motif plays an important role in regulating FtsZ ring dynamics. Our findings uncover a type of amphiphilic beta-strand motif with weak membrane-binding activity and point to the importance of this motif for the dynamic regulation of protein complex formation.

NIGT1.4 maintains primary root elongation in response to salt stress through induction of ERF1 in Arabidopsis.

Plants employ various molecular mechanisms to maintain primary root elongation upon salt stress. Identification of key functional genes, therein, is important for improving crop salt tolerance. Through analyzing natural variation of the primary root length of Arabidopsis natural population under salt stress, we identified NIGT1.4, encoding an MYB transcription factor, as a novel contributor to maintained root growth under salt stress. Using both T-DNA knockout and functional complementation, NIGT1.4 was confirmed to have a role in promoting primary root growth in response to salt stress. The expression of NIGT1.4 in the root was shown induced by NaCl treatments in an ABA-dependent manner. SnRK2.2 and 2.3 were shown to interact with and phosphorylate NIGT1.4 individually. The growth of the primary root of snrk2.2/2.3/2.6 triple mutant was shown sensitive to salt stress, which was similar to nigt1.4 plants. Using DNA affinity purification sequencing, ERF1, a known positive regulator for primary root elongation and salt tolerance, was identified as a target gene for NIGT1.4. The transcriptional induction of ERF1 by salt stress was shown absent in nigt1.4 background. NIGT1.4 was also confirmed to bind to the promoter region of ERF1 by yeast one-hybrid experiment and to induce the expression of ERF1 by dual-luciferase analysis. All data support the notion that salt- and ABA-elicited NIGT1.4 induces the expression of ERF1 to regulate downstream functional genes that contribute to maintained primary root elongation. NIGT1.4-ERF1, therefore, acts as a signaling node linking regulators for stress resilience and root growth, providing new insights for breeding salt-tolerant crops.

Mechanistic insights into the inhibition of the CRISPR-Cas surveillance complex by anti-CRISPR protein AcrIF13.

Clustered regularly interspaced short palindromic repeats (CRISPRs) and CRISPR-associated (Cas) proteins provide prokaryotes with nucleic acid-based adaptive immunity against infections of mobile genetic elements, including phages. To counteract this immune process, phages have evolved various anti-CRISPR (Acr) proteins which deactivate CRISPR-Cas-based immunity. However, the mechanisms of many of these Acr-mediated inhibitions are not clear. Here, we report the crystal structure of AcrIF13 and explore its inhibition mechanism. The structure of AcrIF13 is unique and displays a negatively charged surface. Additionally, biochemical studies identified that AcrIF13 interacts with the type I-F CRISPR-Cas surveillance complex (Csy complex) to block target DNA recognition and that the Cas5f-8f tail and Cas7.6f subunit of the Csy complex are specific binding targets of AcrIF13. Further mutational studies demonstrated that several negatively charged residues of AcrIF13 and positively charged residues of Cas8f and Cas7f of the Csy complex are involved in AcrIF13-Csy binding. Together, our findings provide mechanistic insights into the inhibition mechanism of AcrIF13 and further suggest the prevalence of the function of Acr proteins as DNA mimics.

A Design Method of Two-Dimensional Subwavelength Grating Filter Based on Deep Learning Series Feedback Neural Network.

Subwavelength grating structure has excellent filtering characteristics, and its traditional design method needs a lot of computational costs. This work proposed a design method of two-dimensional subwavelength grating filter based on a series feedback neural network, which can realize forward simulation and backward design. It was programed in Python to study the filtering characteristics of two-dimensional subwavelength grating in the range of 0.4-0.7 µm. The shape, height, period, duty cycle, and waveguide layer height of two-dimensional subwavelength grating were taken into consideration. The dataset, containing 46,080 groups of data, was generated through numerical simulation of rigorous coupled-wave analysis (RCWA). The optimal network was five layers, 128 × 512 × 512 × 128 × 61 nodes, and 64 batch size. The loss function of the series feedback neural network is as low as 0.024. Meanwhile, it solves the problem of non-convergence of the network reverse design due to the non-uniqueness of data. The series feedback neural network can give the geometrical structure parameters of two-dimensional subwavelength grating within 1.12 s, and the correlation between the design results and the theoretical spectrum is greater than 0.65, which belongs to a strong correlation. This study provides a new method for the design of two-dimensional subwavelength grating, which is quicker and more accurate compared with the traditional method.

Who will benefit from computerized cognitive remediation therapy? Evidence from a multisite randomized controlled study in schizophrenia.

BACKGROUND: Computerized cognitive remediation therapy (CCRT) is generally effective for the cognitive deficits of schizophrenia. However, there is much uncertainty about what factors mediate or moderate effectiveness and are therefore important to personalize treatment and boost its effects. METHOD: In total, 311 Chinese inpatients with Diagnostic and Statistical Manual of Mental Disorders-IV schizophrenia were randomized to receive CCRT or Active control for 12 weeks with four to five sessions per week. All participants were assessed at baseline, post-treatment and 3-month follow-up. The outcomes were cognition, clinical symptoms and functional outcomes. RESULTS: There was a significant benefit in the MATRICS Consensus Cognitive Battery (MCCB) total score for CCRT (F1,258 = 5.62; p = 0.02; effect size was 0.27, 95% confidence interval 0.04-0.49). There were no specific moderators of CCRT improvements. However, across both groups, Wisconsin Card Sort Test improvement mediated a positive effect on functional capacity and Digit Span benefit mediated decreases in positive symptoms. In exploratory analyses younger and older participants showed cognitive improvements but on different tests (younger on Symbol Coding Test, while older on the Spatial Span Test). Only the older age group showed MSCEIT benefits at post-treatment. In addition, cognition at baseline negatively correlated with cognitive improvement and those whose MCCB baseline total score was around 31 seem to derive the most benefit. CONCLUSIONS: CCRT can improve the cognitive function of patients with schizophrenia. Changes in cognitive outcomes also contributed to improvements in functional outcomes either directly or solely in the context of CCRT. Age and the basic cognitive level of the participants seem to affect the cognitive benefits from CCRT.

Analysis of antibiotic treatment of children in a Shanghai tertiary hospital based on point prevalence surveys.

BACKGROUND: Misuse and overuse of antibiotics by physicians in the treatment of children is common in China. This study aimed to reveal the overall use of antibiotics to treat children hospitalized in four types of pediatric wards. METHODS: Seven independent point prevalence surveys (PPSs) were conducted in Shanghai Children's Hospital of Shanghai Jiao Tong University over the period 2012 to 2018. Pediatric ward types were defined general pediatric medical, pediatric surgical, pediatric intensive care units (PICU), and neonatal. RESULTS: A total of 3975 pediatric patients were included in the study, of which 63.9% received at least one dose antibiotic. The top five classes of antibiotics administered were cephalosporins (43.8%, n = 1743), penicillins (13.2%, n = 526), carbapenems (8.7%, n = 347), nitroimidazoles (7.1%, n = 281) and macrolides (6.5%, n = 257). The five most commonly used generic antibiotics were cefuroxime (14.9%, n = 594), ceftriaxone (9.7%, n = 387), cefotaxime (9.0%, n = 358), meropenem (8.1%, n = 320) and ampicillin/sulbactam (6.0%, n = 239). Meropenem was among top five antibiotics prescribed in the general pediatric, PICU and neonatal wards and sixth in the pediatric surgical wards. Of all children on antibiotics, 23.4% received prophylactic treatment, and prophylaxis accounted for 68.1% of indications for treatment in the pediatric surgical wards. CONCLUSIONS: Given that over-treatment with third-generation cephalosporins and carbapenems has been associated with treatment-resistant infections, the prescription of these drugs should be strictly controlled and monitored, and measures should be taken to improve the management of surgical prophylaxis in hospitalized children in China.

[Impact of Ureaplasma urealyticum infection on the MRPS22 protein expression in rat spermatogenic cells and intervening effect of Zhibai Dihuang Decoction].

OBJECTIVE: To investigate the impact of Ureaplasma urealyticum (UU) infection (UUI) on the expression of the mitochondrial ribosomal protein S22 (MRPS22) in rat spermatogenic cells and the intervening effect of Zhibai Dihuang Decoction (ZBDH). METHODS: Forty-five SD male rats were randomly divided into four groups of equal number: normal control, UUI model control, ZBDH and azithromycin, and the UUI model was made by bladder injection of the standard UU strain in the latter three groups. After modeling, the rats in the ZBDH and azithromycin groups were treated intragastrically with ZBDH at 1 g/kg/d and azithromycin at 0.105 g/kg/d respectively, while those in the normal and UUI model control groups with normal saline at 1 ml/kg/d. At 21 days after intervention, all the animals were sacrificed and their testes harvested for observation of the apoptosis and mitochondrial ultrastructure of the spermatogenic cells, measurement of the mitochondrial membrane potential (MMP) by flow cytometry, and determination of the mRNA and protein expressions of MRPS22 by RT-PCR and Western blot, respectively. RESULTS: The apoptotic index of the rat sperma-togenic cells was significantly higher in the UUI model control than in the ZBDH and azithromycin groups (ï¼»11.23 ± 1.65ï¼½ % vs ï¼»6.62 ± 0.49ï¼½ % and ï¼»7.82 ± 0.81ï¼½ %, P < 0.01), but lower in the ZBDH than in the azithromycin group (P < 0.05). The mitochondrial ultrastructure of the spermatogenic cells was markedly improved in the ZBDH and azithromycin groups as compared with that in the model control. The MMP level was remarkably lower in the model control than in the normal control (ï¼»8.77 ± 1.73ï¼½ % vs ï¼»22.33 ± 1.66ï¼½ %, P < 0.01), but higher in the ZBDH (ï¼»18.26 ± 1.32ï¼½ %) than in the model control (P < 0.01) and the azithromycin group (ï¼»15.91 ± 1.69ï¼½ %) (P < 0.01). The mRNA and protein expressions of MRPS22 were significantly lower in the model control (8.02 ± 3.21 and 22.65 ± 5.31) than in the normal control (15.43 ± 2.54 and 33.31 ± 7.09), ZBDH (11.26 ± 3.82 and 33.35 ± 3.96), and azithromycin group (8.79 ± 2.03 and 28.11 ± 4.13) (all P < 0.01), but both higher in the ZBDH than in the azithromycin group (P < 0.01). There was a positive correlation between the MRPS22 protein expression and MMP (r = 0.639, P < 0.01). CONCLUSIONS: UU infection induces the apoptosis of rat spermatogenic cells by inhibiting the mRNA and protein expressions of MRPS22 and reducing the mitochondrial membrane potential, while ZBDH can decrease the apoptosis of spermatogenic cells by improving the mRNA and protein expressions of MRPS22 and enhancing the mitochondrial membrane potential.

Toll-like receptor 9 negatively regulates pancreatic islet beta cell growth and function in a mouse model of type 1 diabetes.

AIMS/HYPOTHESIS: Innate immune effectors interact with the environment to contribute to the pathogenesis of the autoimmune disease, type 1 diabetes. Although recent studies have suggested that innate immune Toll-like receptors (TLRs) are involved in tissue development, little is known about the role of TLRs in tissue development, compared with autoimmunity. We aimed to fill the knowledge gap by investigating the role of TLR9 in the development and function of islet beta cells in type 1 diabetes, using NOD mice. METHODS: We generated Tlr9-/- NOD mice and examined them for type 1 diabetes development and beta cell function, including insulin secretion and glucose tolerance. We assessed islet and beta cell number and characterised CD140a expression on beta cells by flow cytometry. We also tested beta cell function in Tlr9-/- C57BL/6 mice. Finally, we used TLR9 antagonists to block TLR9 signalling in wild-type NOD mice to verify the role of TLR9 in beta cell development and function. RESULTS: TLR9 deficiency promoted pancreatic islet development and beta cell differentiation, leading to enhanced glucose tolerance, improved insulin sensitivity and enhanced first-phase insulin secretory response. This was, in part, mediated by upregulation of CD140a (also known as platelet-derived growth factor receptor-α [PDGFRα]). In the absence of TLR9, induced by either genetic targeting or treatment with TLR9 antagonists, which had similar effects on ontogenesis and function of beta cells, NOD mice were protected from diabetes. CONCLUSIONS/INTERPRETATION: Our study links TLR9 and the CD140a pathway in regulating islet beta cell development and function and indicates a potential therapeutic target for diabetes prevention and/or treatment.

[Efficacy and safety of № I Empirical Prescription for Chronic Prostatitis in the treatment of type ⠢ refractory chronic prostatitis].

OBJECTIVE: To evaluate the efficiency and safety of № I Empirical Prescription for Chronic Prostatitis (№ I EPCP) in the treatment of type Ⅲ refractory chronic prostatitis. METHODS: We randomly assigned 53 cases of type Ⅲ refractory chronic prostatitis with damp-heat and blood stasis to an experimental and a control group to receive № I EPCP at 1 dose per day and saw palmetto extract at 160 mg bid), respectively, all for 8 weeks. Before and after 4 and 8 weeks of treatment, we obtained The National Institute of Health Chronic Prostatitis Symptom Index (NIH-CPSI) scores, Traditional Chinese Medicine Syndrome Scores (TCMSS), maximum urinary flow rate (Qmax), average urinary flow rate (Qavg), Hamilton Depression Rating Scale (HAMD) scores and Hamilton Anxiety Rating Scale (HAMA) scores, and compared them between the two groups of patients. RESULTS: Totally 48 of the patients completed the medication and follow-up, 25 in the experimental and 23 in the control group. Compared with the baseline, the NIH-CPSI scores after 8 weeks of treatment were significantly decreased in the experimental (27.82 ± 7.25 vs 15.46 ± 4.77, P <0.05) and the control group (25.98 ± 6.47 vs 21.06 ± 5.74, P <0.05), and so were the TCMSSs (24.64 ± 9.82 vs 16.42 ± 6.33 and 9.15 ± 3.74, P <0.05, and 23.67 ± 8.73 vs 18.55 ± 5.92 and 13.48 ± 4.45, P <0.05); the Qmax at 8 weeks were dramatically increased in the experimental group (ï¼»18.45 ± 7.81ï¼½ vs ï¼»23.44 ± 8.73ï¼½ ml/s, P <0.05) and the control (ï¼»17.58 ± 6.92ï¼½ vs ï¼»21.26 ± 8.32ï¼½ ml/s, P <0.05), and so was the Qavg (ï¼»11.27 ± 5.33ï¼½ vs ï¼»16.51 ± 7.36ï¼½ ml/s, P <0.05 and ï¼»10.66 ± 5.82ï¼½ vs ï¼»13.44 ± 6.16ï¼½ ml/s, P <0.05); the HAMD scores were remarkably reduced in the experimental group (22.74 ± 6.37 vs 17.62 ± 5.71 and 12.54 ± 5.22, P <0.05) and the control (23.55 ± 7.14 vs 22.34 ± 6.88 and 21.62 ± 5.63, P <0.05), and so were the HAMA scores (21.37 ± 7.15 vs 18.42 ± 6.35 and 14.63 ± 7.11, P <0.05 and 20.54 ± 6.77 vs 19.87 ± 6.24 and 19.42 ± 7.04, P <0.05). No obvious adverse reactions were observed in either of the two groups during the medication. CONCLUSIONS: № I EPCP deserves promotion and clinical application for its definite effectiveness and safety in the treatment of type Ⅲ refractory chronic prostatitis with damp-heat and blood stasis.

Romk1 Knockout Mice Do Not Produce Bartter Phenotype but Exhibit Impaired K Excretion.

Romk knock-out mice show a similar phenotype to Bartter syndrome of salt wasting and dehydration due to reduced Na-K-2Cl-cotransporter activity. At least three ROMK isoforms have been identified in the kidney; however, unique functions of any of the isoforms in nephron segments are still poorly understood. We have generated a mouse deficient only in Romk1 by selective deletion of the Romk1-specific first exon using an ES cell Cre-LoxP strategy and examined the renal phenotypes, ion transporter expression, ROMK channel activity, and localization under normal and high K intake. Unlike Romk(-/-) mice, there was no Bartter phenotype with reduced NKCC2 activity and increased NCC expression in Romk1(-/-) mice. The small conductance K channel (SK) activity showed no difference of channel properties or gating in the collecting tubule between Romk1(+/+) and Romk1(-/-) mice. High K intake increased SK channel number per patch and increased the ROMK channel intensity in the apical membrane of the collecting tubule in Romk1(+/+), but such regulation by high K intake was diminished with significant hyperkalemia in Romk1(-/-) mice. We conclude that 1) animal knockouts of ROMK1 do not produce Bartter phenotype. 2) There is no functional linking of ROMK1 and NKCC2 in the TAL. 3) ROMK1 is critical in response to high K intake-stimulated K(+) secretion in the collecting tubule.

Brief cognitive-behavioural therapy for patients in the community with schizophrenia: randomised controlled trial in Beijing, China.

BackgroundBrief cognitive-behavioural therapy (CBT) is an emerging treatment for schizophrenia in community settings; however, further trials are needed, especially in non-Western countries.AimsTo test the effects of brief CBT for Chinese patients with schizophrenia in the community (trial registration: ChiCTR-TRC-13003709).MethodA total of 220 patients with schizophrenia from four districts of Beijing were randomly assigned to either brief CBT plus treatment as usual (TAU) or TAU alone. Patients were assessed at baseline, post-treatment and at 6- and 12-month follow-ups by raters masked to group allocation.ResultsAt the post-treatment assessment and the 12-month follow-up, patients who received brief CBT showed greater improvement in overall symptoms, general psychopathology, insight and social functioning. In total, 37.3% of those in the brief CBT plus TAU group experienced a clinically significant response, compared with only 19.1% of those in the TAU alone group (P = 0.003).ConclusionsBrief CBT has a positive effect on Chinese patients with schizophrenia in the community.

[Effects of Zhibai Dihuang Decoction on mitochondrial cytochrome oxidase in the spermatogenic cells of rats with ureaplasma urealyticum infection].

OBJECTIVE: To explore the effects of Zhibai Dihuang Decoction (ZDD) on mitochondrial cytochrome oxidase (COX) in the spermatogenic cells of rats with ureaplasma urealyticum (UU) infection. METHODS: From forty 4－5 months old SD rats, 30 were randomly selected for the establishment of the model of testicular UU infection by inoculating the bladder with UU suspension and the other 10 injected with normal saline as controls (group A). At 7 days after inoculation, the rat models of testicular UU infection were treated orally with normal saline (group B), ZDD at 1 g per kg of the body weight per day (group C), and azithromycin at 0.105 g per kg of the body weight per day (group D), respectively, once daily for 21 days. Then all the animals were sacrificed and the epididymal and testicular tissues collected for examination of sperm motility with the color sperm dynamic detection system, measurement of the COX activity with the immunohistochemical DAB method, and determination of the mRNA expressions of COXⅠ and COXⅡ by RT-PCR. RESULTS: Compared with group A, group B showed significant decreases in such sperm parameters as grade a sperm (ï¼»1.03 ± 0.09ï¼½ vs ï¼»0.07 ± 0.03ï¼½ %, P<0.01), grade b sperm (ï¼»2.07 ± 0.52ï¼½ vs ï¼»0.35 ± 0.13ï¼½ %, P<0.01), straight line velocity (VSL) (ï¼»10.95 ± 0.98ï¼½ vs ï¼»6.78 ± 1.05ï¼½ µm/s, P<0.01), curvilinear velocity (VCL) (ï¼»42.03 ± 1.35ï¼½ vs ï¼»38.10 ± 7.65ï¼½ µm/s, P>0.05), average path velocity (VAP) (ï¼»16.22 ± 1.52ï¼½ vs ï¼»10.05 ± 1.80ï¼½ µm/s, P<0.01), and the mRNA expressions of COX Ⅰ (ï¼»2.25 ± 0.24ï¼½ vs ï¼»0.93 ± 0.10ï¼½ %, P<0.01) and Ⅱ (ï¼»6.72 ± 0.37ï¼½ vs ï¼»2.95 ± 0.78ï¼½ %, P<0.01). After treatment, all the parameters were remarkably increased in groups C and D (grade a sperm: ï¼»1.11 ± 0.30ï¼½ and ï¼»0.60 ± 0.19ï¼½%; grade b sperm: ï¼»2.40 ± 0.59ï¼½ and ï¼»1.32 ± 0.27ï¼½ %; VSL: ï¼»12.11 ± 1.62ï¼½ and ï¼»11.47 ± 1.21ï¼½ µm/s; VCL: ï¼»54.30 ± 2.35ï¼½ and ï¼»45.75 ± 1.64ï¼½ µm/s; VAP ï¼»18.40 ± 1.27ï¼½ and ï¼»16.69 ± 1.02ï¼½ µm/s; expression of COXⅠ mRNA: ï¼»1.86 ± 0.30ï¼½ and ï¼»1.74 ± 0.17ï¼½ %) as compared with those in group B (P<0.05or P<0.01) except the COX activity and the expression of COX Ⅱ mRNA (P>0.05), and all the parameters were significantly higher in group C than in D (P<0.05or P<0.01). CONCLUSIONS: UU infection can reduce grades a and b sperm, linear, curvilinear and mean sperm velocities, and the mRNA expressions of COX Ⅰ and Ⅱ while ZDD can improve these parameters. The improvement of sperm motility may not be associated with the activity of COX, and the COX activity may be related to the mRNA expression of COX II but not that of COXⅠ.

A novel "humanized mouse" model for autoimmune hepatitis and the association of gut microbiota with liver inflammation.

UNLABELLED: Autoimmune hepatitis (AIH) in humans is a severe inflammatory liver disease characterized by interface hepatitis, the presence of circulating autoantibodies, and hyper-gammaglobulinemia. There are two types of AIH, type 1 (AIH-1) and type 2 (AIH-2), characterized by distinct autoimmune serology. Patients with AIH-1 are positive for anti-smooth muscle and/or antinuclear autoantibodies, whereas patients with AIH-2 have anti-liver kidney microsomal type 1 and/or anti-liver cytosol type 1 autoantibodies. Cytochrome P4502D6 is the antigenic target of anti-liver kidney microsomal type 1, and formiminotransferase cyclodeaminase is the antigenic target of anti-liver cytosol type 1. It is known that AIH, both types 1 and 2, is strongly linked to the human leukocyte antigen (HLA) alleles -DR3, -DR4, and -DR7. However, direct evidence of the association of HLA with AIH is lacking. We developed a novel mouse model of AIH using the HLA-DR3 transgenic mouse on the nonobese-diabetic background by immunization of HLA-DR3- and HLA-DR3+ nonobese-diabetic mice with a DNA plasmid, coding for human cytochrome P4502D6/formiminotransferase cyclodeaminase fusion protein. Immunization with cytochrome P4502D6/formiminotransferase cyclodeaminase leads to a sustained elevation of alanine aminotransferase, development of antinuclear autoantibodies and anti-liver kidney microsomal type 1/anti-liver cytosol type 1 autoantibodies, chronic immune cell infiltration, and parenchymal fibrosis on liver histology in HLA-DR3+ mice. Immunized mice also showed an enhanced T helper 1 immune response and paucity of the frequency of regulatory T cells in the liver. Moreover, HLA-DR3+ mice with exacerbated AIH showed reduced diversity and total load of gut bacteria. CONCLUSION: Our humanized animal model has provided a novel experimental tool to further elucidate the pathogenesis of AIH and to evaluate the efficacy and safety of immunoregulatory therapeutic interventions in vivo.

[Effect of Zhibai Dihuang Decoction on Expressions of TRPV1 and TRPV5 in Spermatogenic Cells of UU-infected Rats].

OBJECTIVE: To observe the effect of Zhibai Dihuang Decoction (ZDD) on mRNA and protein expressions of transient receptor potential family vanilloid subtype 1 (TRPV1) and transient receptor potential family vanilloid subtype 5 (TRPV5) in Ureaplasma urealyticum (UU)-infected rat semens and spermatogenic cells, and to explore the pathomechanism of UU-infected infertility and the intervention of ZDD. METHODS: Totally 45 were randomly selected from 60 4-5 months old SD rats. UU testicular infected animal models were set up after bladder inoculation of UU suspension. The remaining 15 rats were simultaneously injected with normal saline as a normal control group. After a successful modeling, UU infected model rats were randomly divided into the model group, the azithromycin group, and the ZDD group, 15 in each group. Rats in the ZDD group were administered with ZDD at the daily dose of 1 g/kg by gastrogavage. Rats in the azithromycin group were administered with azithromycin suspension at the daily dose of 0. 105 mg/kg by gastrogavage. Equal volume of normal saline was administered to rats in the normal control group and the model group. All medication was performed once daily for 21 successive days. Testes and epididymis were extracted after rats were killed and UU positive rates were compared among all groups. Sperm cells were separated using a mechanical separation technique. Sperm motility parameters were detected using color sperm motion detection system. mRNA and protein expressions of TRPV1 and TRPV5 in spermatogenic cells were determined by real-time quantitative PCR and Western blot. RESULTS: The UU positive rate was obviously higher in the model group than in the normal control group [(86.7% (13/15 cases) vs. 0] P < 0.05). It was lower in the ZDD group [33.3% (5/15 cases)] and the azithromycin group [26.7% (4/15 cases)] than in the model group (P < 0.05). Compared with the normal control group, class A and B sperms were reduced, the linear velocity and the average velocity were significantly lowered, mRNA and protein expressions of TRPV1 and TRPV5 in spermated genic cells significantly decreased in the model group (P < 0.05). Compared with the model group, class A and B sperms were increased, linear and curve velocities and the average velocity were significantly elevated, mRNA and protein expressions of TRPV1 and TRPV5 significantly increased in the ZDD group and the azithromycin group (P < 0.05, P < 0.01). Compared with azithromycin group, class A and B sperms were increased, the linear velocity and the average velocity were significantly elevated, mRNA and protein expressions of TRPV1 and TRPV5 significantly increased in the ZDD group (P < 0.05, P < 0.01). CONCLUSION: ZDD could fight against UU infection and elevate semen quality, which might be associated with up-regulating mRNA and protein expressions of TRPV1 and TRPV5 in spermatogenic cells.

Type-specific detection of 30 oncogenic human papillomaviruses by genotyping both E6 and L1 genes.

Human papillomavirus (HPV) is the principal cause of invasive cervical cancer and benign genital lesions. There are currently 30 HPV types linked to cervical cancer. HPV infection also leads to other types of cancer. We developed a 61-plex analysis of these 30 HPV types by examining two genes, E6 and L1, using MassARRAY matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) (PCR-MS). Two hundred samples from homosexual males (HM) were screened by PCR-MS and MY09/MY11 primer set-mediated PCR (MY-PCR) followed by sequencing. One hundred thirty-five formalin-fixed, paraffin-embedded (FFPE) cervical cancer samples were also analyzed by PCR-MS, and results were compared to those of the commercially available GenoArray (GA) assay. One or more HPV types were identified in 64.5% (129/200) of the samples from HM. Comprising all 30 HPV types, PCR-MS detected 51.9% (67/129) of samples with multiple HPV types, whereas MY-PCR detected only one single HPV type in these samples. All PCR-MS results were confirmed by MY-PCR. In the cervical cancer samples, PCR-MS and GA detected 97% (131/135) and 90.4% (122/135) of HPV-positive samples, respectively. PCR-MS and GA results were fully concordant for 122 positive and 4 negative samples. The sequencing results for the 9 samples that tested negative by GA were completely concordant with the positive PCR-MS results. Multiple HPV types were identified in 25.2% (34/135) and 55.6% (75/135) of the cervical cancer samples by GA and PCR-MS, respectively, and results were confirmed by sequencing. The new assay allows the genotyping of >1,000 samples per day. It provides a good alternative to current methods, especially for large-scale investigations of multiple HPV infections and degraded FFPE samples.

Prevalence of 10 human polyomaviruses in fecal samples from children with acute gastroenteritis: a case-control study.

We conducted a case-control study to explore the prevalence of 10 human polyomaviruses in fecal specimens from hospitalized children with diarrhea and asymptomatic control subjects by using multiplex PCR detected by matrix-assisted laser desorption ionization-time of flight mass spectrometry. The differences between cases and controls were not statistically significant.

Acquired ALK G1202R-, ALK I1171N-, or EML4-ALK-mediated resistance to ensartinib in lung adenocarcinoma but responded to lorlatinib: A case report.

ALK rearrangements are identified as driver mutations in non-small-cell lung cancer (NSCLC). EML4 is the most common partner of ALK rearrangements. Here, we reported a patient with lung adenocarcinoma who was identified with EML4-ALK mutations when he progressed on an immune checkpoint inhibitor. The patient was treated with alectinib and obtained a progression-free survival (PFS) of 24 months. Then, next-generation sequencing on circulating tumor DNA identified multiple ALK mutations, including ALK G1202R, I1171N, ALK-ENC1, and EML4-ALK. Ensartinib was given, and the patient achieved a PFS of 5 months. After progression, lorlatinib was administered, and the patient achieved a partial response. Now, the benefit is still ongoing with a PFS over 10 months. Our case may provide evidence for the treatment choice of multiple ALK mutations, including ALK I1171N.

Taxi drivers' traffic violations detection using random forest algorithm: A case study in China.

OBJECTIVE: To effectively explore the impacts of several key factors on taxi drivers' traffic violations and provide traffic management departments with scientific decisions to reduce traffic fatalities and injuries. METHODS: 43,458 electronic enforcement data about taxi drivers' traffic violations in Nanchang City, Jiangxi Province, China, from July 1, 2020, to June 30, 2021, were utilized to explore the characteristics of traffic violations. A random forest algorithm was used to predict the severity of taxi drivers' traffic violations and 11 factors affecting traffic violations, including time, road conditions, environment, and taxi companies were analyzed using the Shapley Additionality Explanation (SHAP) framework. RESULTS: Firstly, the ensemble method Balanced Bagging Classifier (BBC) was applied to balance the dataset. The results showed that the imbalance ratio (IR) of the original imbalanced dataset reduced from 6.61% to 2.60%. Moreover, a prediction model for the severity of taxi drivers' traffic violations was established by using the Random Forest, and the results showed that accuracy, m_F1, m_G-mean, m_AUC, and m_AP obtained 0.877, 0.849, 0.599, 0.976, and 0.957, respectively. Compared with the algorithms of Decision Tree, XG Boost, Ada Boost, and Neural Network, the performance measures of the prediction model based on Random Forest were the best. Finally, the SHAP framework was used to improve the interpretability of the model and identify important factors affecting taxi drivers' traffic violations. The results showed that functional districts, location of the violation, and road grade were found to have a high impact on the probability of traffic violations; their mean SHAP values were 0.39, 0.36, and 0.26, respectively. CONCLUSIONS: Findings of this paper may help to discover the relationship between the influencing factors and the severity of traffic violations, and provide a theoretical basis for reducing the traffic violations of taxi drivers and improving the road safety management.