Objective To construct a non-trace deletion mutant of exlA in Pseudomonas aeruginosa strain NY8755(NY8755ΔexlA)and investigate the basic characteristics of pore-forming toxin ExlA.Methods The NY8755ΔexlA was constructed using the secondary homologous recombination method.C57BL/6J female mice ages 6 to 8 weeks were infected with NY8755 and NY8755 ΔexlA via aerosolized intratraheal inoculation respectively.Within 7 days of infection,the survival and weight changes of the mice were observed and recorded before the proinflammatory cytokines in the bronchoal-veolar lavage fluid(BALF)of the infected mice in the two groups were detected.Results The sequencing results showed that NY8755 ΔexlA was constructed.After 1×107 CFU NY8755 and NY8755 ΔexlA were infected,all the mice in the wild-type strain group died within 48 hours,while those in the mutant strain group began to die after 48 hours,and 40%of them remained alive 7 days later.The weight of surviving mice in the mutant strain group decreased but recovered gradually.After 12 hours of infection,there were more bloody exudates(redder in color)in the BALF of the wild-type strain group than in the mutant strain group,and the contents of proinflammatory cytokines interleukin-1β(IL-1β)and interleukin-17A (IL-17A)were significantly different. Conclusion Pseudomonas aeruginosa pore-forming toxin ExlA is the key pathogenic virulence factor of the exlA-positive Pseudomonas aeruginosa,which can significantly affect the survival status of mice and cause obvious inflammation in mice. Very little information is available on the action mechanisms of ExlA. In this study, The NY8755ΔexlA and the C57BL/6J mouse models infected with NY8755 and NY8755ΔexlA have been constructed that may be used for the investigation of pathogenesis of exlA-positive Pseudomonas aeruginosa.

Objective:To determine S100A10 protein expression in psoriatic lesions, and to investigate its effect on psoriasis-like skin inflammation in imiquimod (IMQ) -induced mouse models.Methods:From January 2020 to June 2022, skin lesions were surgically collected from 28 patients with psoriasis in the Department of Dermatology, the First Affiliated Hospital of Xinxiang Medical University; normal skin tissues were collected from 18 healthy subjects in the Department of General Surgery and Department of Ophthalmology during the same period, and served as a control group. Immunohistochemical staining was performed to determine the S100A10 protein expression in skin lesions from psoriasis patients and normal skin tissues from healthy controls. Ten wild-type (WT) C57BL/6J female mice and 10 S100A10 -/- C57BL/6J female mice were selected to establish the IMQ-induced mouse models of psoriasis-like skin inflammation. Then, the mice were randomly divided into gene knock-out (KO) /IMQ group, WT/IMQ group, KO/vaseline (VAS) group, and WT/VAS group by using a random number table, and there were 5 mice in each group. The mice in the KO/IMQ group and WT/IMQ group were topically treated with IMQ cream (62.5 mg) on the shaved back daily to establish the mouse models of psoriasis-like skin inflammation, while the mice in the KO/VAS group and WT/VAS group were topically treated with vaseline cream (62.5 mg) daily, and both treatments lasted 7 consecutive days. Skin lesions on the back were observed daily. On day 7, the mice were sacrificed by cervical dislocation, and their dorsal skin tissues were excised. The IMQ-induced psoriasis-like skin inflammation was evaluated by the psoriasis area and severity index (PASI), pathological manifestations of skin lesions were observed by hematoxylin and eosin staining, the expression of S100A10 and Ki-67 in skin lesions was determined by immunohistochemical staining, the expression of STAT3, IL-17A and other cytokines was determined by Western blot analysis, and IL-17 mRNA expression was determined by real-time fluorescence-based quantitative PCR (qPCR). Statistical analysis was carried out by using the independent sample t-test, nonparametric U test, and chi-square test. Results:Immunohistochemical staining showed that the S100A10 protein expression was significantly lower in the psoriasis vulgaris lesions than in the normal control skin tissues ( Z = -3.47, P < 0.001). In the mouse models, the S100A10 protein expression was significantly lower in the skin lesions of mice in the WT/IMQ group than in the skin tissues of mice in the WT/VAS group ( t = 3.64, P = 0.007), and the Ki-67 expression was significantly higher in the KO/IMQ group than in the WT/IMQ group ( t = 2.97, P = 0.041). Additionally, the mice in the KO/IMQ group presented with more severe clinical manifestations such as scales and infiltration compared with those in the WT/IMQ group. Western blot analysis showed that the phosphorylated STAT3/STAT3 expression and IL-17A protein expression was significantly higher in the KO/IMQ group than in the WT/IMQ group ( t = 3.27, 3.48, P = 0.031, 0.025, respectively), and qPCR revealed that the IL-17A mRNA expression was also significantly higher in the KO/IMQ group than in the WT/IMQ group ( t = 2.73, P = 0.029) . Conclusion:S100A10 protein was underexpressed in the skin lesions of psoriasis patients, and the deletion of S100A10 protein aggravated IMQ-induced psoriasis-like skin inflammation in mice, possibly by upregulating STAT3 phosphorylation in the epidermis.

Objective:To investigate the effect of cesarean scar defect and endometrial cavity fluid on the pregnancy outcomes of infertility patients with previous cesarean scar uterus undergoing in vitro fertilization-embryo transfer (IVF-ET). Methods:This was a retrospective cohort study. Totally 732 cases of patients with previous cesarean scar uterus were selected from frozen-thawed embryo transfer (FET) cycles in the Fertility Center, Shenzhen Zhongshan Urology Hospital from January 2019 to March 2020. They were divided into four groups: group A ( n=39) including patients with previous cesarean scar defect and without endometrial cavity fluid; group B ( n=82) including patients with previous cesarean scar defect and with endometrial cavity fluid; group C ( n=495) including patients without previous cesarean scar defect and without endometrial cavity fluid; group D ( n=116) including patients without previous cesarean scar defect and with endometrial cavity fluid. The general data and pregnancy outcomes were compared among these groups. Multivariate logistics regression analysis of pregnancy outcome indexes was performed. Results:The transplantation age of group A was higher than that of group B [(38.33±3.55) years vs. (36.93±3.59) years, P=0.045], the endometrial thickness of luteal transformation day and the rate of good-quality embryo transplantation of group C were higher than those of group D [(9.40±1.56) mm vs. (9.03±1.59) mm, P=0.025; 75.76% (375/495) vs. 65.52% (76/116), P=0.024]. The egg retrieval age of group A was higher than that of group C [(37.72±3.55) years vs. (36.25±4.52) years, P=0.049], but the endometrial thickness of luteal transformation day was thinner [(8.74±1.58) mm vs. (9.40±1.56) mm, P=0.012], and the differences were statistically significant. The implantation rate, the biochemical pregnancy rate and the clinical pregnancy rate of group C were higher than those of group D [34.23% (230/672) vs. 22.58% (35/155), P=0.007; 48.28% (239/495) vs. 37.93% (44/116), P=0.044; 42.83% (212/495) vs. 30.17% (35/116), P=0.012]. The early abortion rate in group D was higher than that in group B [40.00% (14/35) vs. 17.24 % (5/29), P=0.047], the difference was statistically significant. Multivariate logistics regression analyses were used with adjustment for possible confounders: the maternal age at embryo transfer, the maternal age at egg retrieval, anti-Müllerian hormone (AMH),antral follicle count (AFC), basic follicle-stimulating hormone (FSH), endometrial CD138 results, FET protocol, and embryo attributes, the number of embryos transferred, embryo quality, the endometrial thickness and the progesterone value on the day of luteal transformation, the result showed that the implantation rate, the biochemical pregnancy rate and the clinical pregnancy rate of group D were lower than those of group C [22.58% (35/155) vs. 34.23% (230/672), P=0.006; 37.93% (44/116) vs. 48.28% (239/495), P=0.047; 30.17% (35/116) vs. 42.83% (212/495), P=0.022] and the differences were statistically significant. The implantation rate, the biochemical pregnancy rate and the clinical pregnancy rate of group B were higher than those of group D [29.52% (31/105) vs. 22.58% (35/155), P=0.049; 48.78% (40/82) vs. 37.93% (44/116), P=0.012; 35.37% (29/82) vs. 30.17% (35/116), P=0.030] and the differences were statistically significant. Conclusion:Endometrial cavity fluid is the main factor that obviously affects the pregnancy outcome of infertility patients with previous cesarean scar uterus undergoing FET cycles.

Objective:To investigate the effect of cesarean scar defect and endometrial cavity fluid on the pregnancy outcomes of infertility patients with previous cesarean scar uterus undergoing in vitro fertilization-embryo transfer (IVF-ET). Methods:This was a retrospective cohort study. Totally 732 cases of patients with previous cesarean scar uterus were selected from frozen-thawed embryo transfer (FET) cycles in the Fertility Center, Shenzhen Zhongshan Urology Hospital from January 2019 to March 2020. They were divided into four groups: group A ( n=39) including patients with previous cesarean scar defect and without endometrial cavity fluid; group B ( n=82) including patients with previous cesarean scar defect and with endometrial cavity fluid; group C ( n=495) including patients without previous cesarean scar defect and without endometrial cavity fluid; group D ( n=116) including patients without previous cesarean scar defect and with endometrial cavity fluid. The general data and pregnancy outcomes were compared among these groups. Multivariate logistics regression analysis of pregnancy outcome indexes was performed. Results:The transplantation age of group A was higher than that of group B [(38.33±3.55) years vs. (36.93±3.59) years, P=0.045], the endometrial thickness of luteal transformation day and the rate of good-quality embryo transplantation of group C were higher than those of group D [(9.40±1.56) mm vs. (9.03±1.59) mm, P=0.025; 75.76% (375/495) vs. 65.52% (76/116), P=0.024]. The egg retrieval age of group A was higher than that of group C [(37.72±3.55) years vs. (36.25±4.52) years, P=0.049], but the endometrial thickness of luteal transformation day was thinner [(8.74±1.58) mm vs. (9.40±1.56) mm, P=0.012], and the differences were statistically significant. The implantation rate, the biochemical pregnancy rate and the clinical pregnancy rate of group C were higher than those of group D [34.23% (230/672) vs. 22.58% (35/155), P=0.007; 48.28% (239/495) vs. 37.93% (44/116), P=0.044; 42.83% (212/495) vs. 30.17% (35/116), P=0.012]. The early abortion rate in group D was higher than that in group B [40.00% (14/35) vs. 17.24 % (5/29), P=0.047], the difference was statistically significant. Multivariate logistics regression analyses were used with adjustment for possible confounders: the maternal age at embryo transfer, the maternal age at egg retrieval, anti-Müllerian hormone (AMH),antral follicle count (AFC), basic follicle-stimulating hormone (FSH), endometrial CD138 results, FET protocol, and embryo attributes, the number of embryos transferred, embryo quality, the endometrial thickness and the progesterone value on the day of luteal transformation, the result showed that the implantation rate, the biochemical pregnancy rate and the clinical pregnancy rate of group D were lower than those of group C [22.58% (35/155) vs. 34.23% (230/672), P=0.006; 37.93% (44/116) vs. 48.28% (239/495), P=0.047; 30.17% (35/116) vs. 42.83% (212/495), P=0.022] and the differences were statistically significant. The implantation rate, the biochemical pregnancy rate and the clinical pregnancy rate of group B were higher than those of group D [29.52% (31/105) vs. 22.58% (35/155), P=0.049; 48.78% (40/82) vs. 37.93% (44/116), P=0.012; 35.37% (29/82) vs. 30.17% (35/116), P=0.030] and the differences were statistically significant. Conclusion:Endometrial cavity fluid is the main factor that obviously affects the pregnancy outcome of infertility patients with previous cesarean scar uterus undergoing FET cycles.

Objective This study aims to investigate the correlation of CD4+ CD25+ FOXP3+ Tregs and PD‐1+ CD4+ T cells located at cervical lesions and the prognosis of CIN .PD‐1 expression on T cells induces Tregs .Methods Cervical lymphocytes were collected from CIN patients and analyzed by FACS .Comparisons were made between populations of cervical Tregs and PD‐1+CD4+T cells in CIN regressors and non‐regressors .Results A median of 11 .8% of cervical CD4+ T cells were CD4+ CD25+ FOXP3+Tregs ,while a median of 30 .3% were PD‐1+ CD4+ T cells .The proportions of CD4+ CD25+ FOXP3+ Tregs and PD‐1+ cells were significantly lower in CIN regressors when compared with non‐regressors .Conclusion The prevalence of CD4+ CD25+ FOXP3+Tregs negative correlates with spontaneous regression of CIN ,suggesting that cervical Tregs may play an important role in HPV‐related neoplastic immunoevasion .

Objective: To investigate the gene frequency of HLA-B* 5801 in the population of Chinese Minnan region.Methods:In this study,we enrolled 178 patients requiring allopurinol therapy( including 40 patients with gout,89 patients with hyperuricemia and 49 patients with gouty arthritis) and 100 healthy people.We isolated genomic DNA from their blood and screened for HLA-B*5801 with both PCR and gene sequencing.Results:We found 22%patients and 16%healthy people with HLA-B*5801.The frequencies of HLA-B*5801 in patients and healthy people are 0.13 and 0.09,respectively.The results from PCR and gene sequencing were consistent.Conclusion:The frequency of HLA-B*5801 in the population of Chinese Minnan region is relatively high.Therefore,it is necessary to screen for HLA-B*5801 in allopurinol users before taking the medicine.

The 10-month baby boy,with normal development,mainly due to sleep in frequent tongue bite nearly 4 months.Bitten his tongue after faring asleep,biting bleeding,bite pain awake.Many of his tongue ulcers,serious impact on children's lives,family companionship in suffering.History found in the supplementary week before the onset of the left frontal children hurt skin bruising.Electroencephalogram showed:Sleep of epileptiform discharges in the left frontal and central anterior temporal areas,but bite the tongue during sleep electroencephalogram synchronization no relevant abnormal discharge.The final diagnosis of traumatic epilepsy,frontal lobe epilepsy syndrome automatically lead to tongue bite tongue with traumatic ulcers.Oral Clonazepam 0.25 mg before sleep,the symptoms disappeared that night,nighttime sleep peacefully.His tongue ulceration has healed after a month.Readers are advised to take advantage of these key parts of the diagnostic process and diagnostic thinking or diagnostic procedures,combined with their own clinical practice,serious thinking,learning,summarized,and benefit from it.

OBJECTIVEThe authors sought to investigate the clinical features and characteristics of genetic mutation in patients with aspartylglucosaminuria.

METHODClinical data of two pediatric siblings in a family were analyzed retrospectively and relative literature was reviewed in order to study the clinical features, imaging and enzymatic characteristics and genetic mutations.

RESULTCase 1, the proband, male, he was hospitalized at 20 months of age because of fever and hepatosplenomegaly for nine days. This child was of moderate nutritional status and normal development. Blood tests showed hemoglobin 78.0 g/L, RBC3.18 × 10¹²/L, WBC 4.06 × 10⁹/L, neutrophils 0.236, lymphocytes 0.631, platelets 34 × 10⁹/L, C-reactive protein 17 mg/L. Blood biochemistry showed alanine aminotransferase 67.1 U/L, aspartate aminotransferase 74.1 U/L, serum albumin 32.8 g/L, direct bilirubin 10.5 µmol/L, lactate dehydrogenase 301.7 U/L. Bone marrow cytology showed reactive morphological changes in bone marrow cells. Atypical lymphocytes could be seen in both peripheral blood and bone marrow smears. Cranial MRI showed poor myelination. Aspartylglucosaminidase activity in peripheral leucocytes of the proband 5.7 nmol/(g × min) vs. normal control>26.6 nmol/(g × min). On his AGA gene and that of his parents, a heterozygous mutation site located in exon 3, c.392C>T (p.S131L), was identified as a novel mutation inherited from his father. The mutation from his mother has not been detected. The proband was not responsive to the anti-infectious medication, nutritional intervention and symptomatic treatment.He died one month after diagnosis.His elder brother, Case 2, showed fever, recurrent respiratory tract infection and progressive psychomotor regression with hepatosplenomegaly from the age of four years. Cranial MRI revealed extensive symmetrical leukodystrophy in bilateral cerebra, cerebellum and brainstem.He died at the age of six years.Related literature was summarized, and no Chinese AGU cases had been reported; 221 foreign cases were collected. The clinical and imaging characteristics were summarized. Delay in language development was one of the clinical symptoms that the majority of parents of AGU children first noted.

CONCLUSIONPatients with aspartylglucosaminuria lack of specific symptoms.For children with unexplained delayed speech and progressive mental retardation, the possibility of AGU should be considered, and efforts be made for enzymatic and genetic diagnosis. c.392C> T (p.S131L) was identified as a novel mutation of AGA gene.

Aspartylglucosaminuria ; diagnosis ; genetics ; pathology ; Aspartylglucosylaminase ; genetics ; metabolism ; Biomarkers ; blood ; Brain ; pathology ; Child, Preschool ; DNA Mutational Analysis ; Heterozygote ; Humans ; Infant ; Lysosomal Storage Diseases ; diagnosis ; genetics ; pathology ; Magnetic Resonance Imaging ; Male ; Mutation ; Pedigree ; Polymerase Chain Reaction

OBJECTIVETo evaluate the therapeutic effect and safety of rapamycin in treatment of children with tuberous sclerosis complex (TSC) complicated with epilepsy.

METHODThis was an open-label, prospective, self-controlled study. From Sep. 2011 to Sep. 2013, 52 patients with the diagnosis of tuberous sclerosis complicated with epilepsy receiving rapamycin treatment for at least 24 weeks were enrolled.

RESULTOf the 52 children, 34 were male and 18 female. The median age at onset of epilepsy was 4.8 months (4 days-49 months), the median age for treatment with rapamycin was 27 months (4.5-172.5 months). Ten children had a family history of TSC. In 24 children TSC gene detection was carried out, among whom TSC1 mutation was detected in 4 cases and TSC2 mutation in 20. Before rapamycin therapy, 59.62%, (31/52) patients took more than 3 antiepileptic drugs, of whom 10 cases even took more than 5 kinds of antiepileptic drugs. Fifty-two patients received rapamycin treatment for 24 weeks, seizure free rate was 25.00% (13 cases), the total effective rate was 73.08% (38 cases); 31 cases received treatment for 48 weeks, seizure free 6 cases, total effective 23 cases; 17 cases accepted treatment for 72 weeks, seizure free 5 cases, total effective 13 cases; 12 cases received treatment for 96 weeks, seizure free 3 cases, total effective 9 cases. With the decrease of seizure attacks, use of antiepileptic drug types were reduced simultaneously, they had a negative correlation. Before rapamycin therapy, the average frequency of seizures was 70.27 times/d, the number of antiepileptic drug kinds was 1.30. After 24, 48, 72, 96 weeks' treatment, the average seizure frequency was reduced to 1.94-2.80 times /d and the antiepileptic drugs were reduced to 0.83-0.97 kinds. On every visit during the follow-up, blood and urine routine tests, liver and kidney function test showed no abnormality in the 52 cases. The drug dosage was 1 mg/(m(2)×d), average 0.7 mg/d (0.35-1.20 mg/d). Blood concentrations of rapamycin remained below 10 µg/L (average 6.5 µg/L). The main side effect was oral ulcer which happened in 23.08% (12/52). The oral ulcer would disappeared 2-3 days later. 17.31% (9/52 cases) had upper respiratory infection.

CONCLUSIONRapamycin was effective in children with tuberous sclerosis and epilepsy with few adverse reactions. The daily dose of rapamycin for children patients is 1 mg/m(2), which has a certain effect on seizures and a good safety profile.

Adolescent ; Anticonvulsants ; adverse effects ; therapeutic use ; Child ; Child, Preschool ; Epilepsy ; complications ; drug therapy ; Female ; Humans ; Infant ; Male ; Prospective Studies ; Seizures ; prevention & control ; Sirolimus ; adverse effects ; therapeutic use ; Treatment Outcome ; Tuberous Sclerosis ; complications ; genetics

Objective To investigate the distribution of carbapenemase-resistant genes carried by multi-drug resistant Acineto-bacter baumannii .Methods 80 strains of multi-drug resistant Acinetobacter baumannii were collected .Polymerase chain reaction (PCR)wasusedtodetectcarbapenemase-resistantgenes,suchasOXA-23,OXA-24,OXA-51,OXA-58,SIM,IMP,VIM ,GIMand SPM ,in multi-drug resistant Acinetobacter baumannii .Results Drug resistant gene OXA-23 [49 (61 .3% )] ,OXA-51 [73 (91 .3% )] ,OXA-58[7(8 .8% )] ,OXA-24[1(1 .3% )] ,IMP[17(21 .3% )] and VIM[2(2 .5% )] were found in 80 strains of multi-drug resistant Acinetobacter baumannii ,while GIM ,SIM and SPM gene were not found .Conclusion IMP ,OXA ,VIM is the main genotypes carried by multi-drug resistant Acinetobacter baumannii .