Objective:To explore the effect of specific inhibitor of Smad3 (SIS3) on glucocorticoid-induced ocular hypertension in mice and its possible mechanism.Methods:Fifty-one eight-week-old female C57BL/6J mice were randomly divided into control group, dexamethasone group and SIS3 group by the random number table method, with 17 mice in each group.Mice in the control group were injected with 20 μl 2 % polyvinyl alcohol into the conjunctival fornix every week for 4 weeks.Mice in the dexamethasone group and SIS3 group were injected with 20 μl 10 mg/ml dexamethasone acetate every week and SIS3 group was treated with additional 100 μg/ml SIS3 nanomicelle eye drops 3 times daily for up to 4 weeks.Intraocular pressure (IOP) was measured weekly using Icare rebound tonometer.Mice were sacrificed 4 weeks after treatment, and the eyeballs were removed.Morphology of trabecular meshwork (TM) tissues were detected by hematoxylin-eosin (HE) staining.The collagen deposition area in TM tissues were examined by Masson staining.Fibronectin (FN) and collagen type Ⅰ (Col-1) in the extracellular matrix of TM tissue were detected by immunofluorescence staining.TM tissues were obtained from donated patients, and primary human trabecular meshwork cells (HTMCs) were obtained by culture.The expression level of myocilin in dexamethasone-induced HTMCs was detected by immunofluorescence and Western blot for cell identification.Primary HTMCs were divided into normal control group, dexamethasone group and SIS3 group cultured with normal culture medium, medium containing 400 nmol/L dexamethasone, medium containing 400 nmol/L dexamethasone+ 10 μmol/L SIS3 for 48 hours, respectively.The expression levels of FN, Col-1 and p-Smad3/Smad3 proteins were measured by Western blot.The use and care of animals complied with the ARVO statement.This study protocol was approved by the Animal Ethics Committee of Zhengzhou University (No.ZZU-LA20220729).The collection of TM tissue specimens complied with the Declaration of Helsinki and was approved by the Medical Ethics Committee of Henan Provincial Eye Hospital (No.HNEECKY-2022[18]).The patients knew the purpose of the experiment and signed the informed consent forms.Results:There was a significant overall difference in IOP among the three groups at different time points after administration ( Fgroup=72.94, P<0.001; Ftime=33.19, P<0.001).Compared with baseline, IOP was increased in the dexamethasone group at each time point after administration, and the differences were statistically significant (all P<0.001).The IOP of the control and SIS3 groups at weeks 1, 2, 3, 4 were significantly lower than that of the dexamethasone group (all P<0.001).HE staining showed that the iridocorneal angles of all groups were open with similar morphology of the TM structure.Masson staining showed that the positive expression area of collagen in the control group, dexamethasone group and SIS3 group was (9.57±2.91)%, (27.75±5.88)% and (11.67±3.78)%, respectively, with a statistically significant difference among the three groups ( F=25.91, P<0.001), and the positive expression area of collagen was significantly lower in the control group and SIS3 group than in the dexamethasone group (all P<0.001).The fluorescence expression level of FN in the control group, dexamethasone group and SIS3 group was 8.00±1.92, 14.01±2.74 and 7.85±0.64, respectively, and the fluorescence expression level of Col-1 was 6.90±1.16, 14.36±3.19 and 4.90±0.88, respectively, with statistically significant differences among the three groups ( F=15.93, 30.29; both P<0.001), and the fluorescence expression levels of FN and Col-1 were significantly lower in the control group and SIS3 group than in the dexamethasone group (all P<0.01).Immunofluorescence staining and Western blot showed that the cultured primary cells expressed myocilin and the expression level of myocilin was significantly increased after dexamethasone induction, which was identified as HTMCs.There were statistically significant differences in the relative expression levels of FN, Col-1, and p-Smad3/Smad3 proteins among different groups of cells ( F=8.22, 23.08, 8.78; all P<0.05), and the relative expression levels of FN, Col-1, and p-Smad3/Smad3 proteins were significantly lower in the control group and SIS3 group than in the dexamethasone group (all P<0.05). Conclusions:SIS3 reduces IOP by inhibiting p-Smad3, reducing extracellular matrix deposition in TM, and reducing fibrosis in the TM tissue.

Objective:To search, evaluate, and summarize the best evidence of built-in fecal incontinence management device, to inform the management of incontinence dermatitis by clinical healthcare professionals.Methods:BMJ Best Practice, UpToDate, Guideline International Network, Joanna Briggs Institute, National Institute for Health and Care Excellence, Scottish Intercollegiate Guidelines Network, Registered Nurses′Association of Ontario, The Cochrane Library, Medline, Embase, SinoMed, CINAHL, PubMed, Web of Science, OVID, China National Knowledge Infrastructure, Wanfang Database were systematically searched for all evidence regarding the application of fecal collection devices. It included clinical practice, guidelines, systematic reviews, expert consensuses, evidence summaries, and randomized controlled trial. Two researchers independently evaluated the literature quality and extracted the literature that met the standards.Results:A total of 12 pieces of the literature were involved, including 2 best practice, 5 guidelines, 3 expert consensuses, and 2 systematic reviews. This study summarized 26 pieces of best evidence in relation to the following 5 themes: indications and contraindications, device insertion, device maintenance, device removal and effectiveness evaluation.Conclusions:This study scientifically and systematically summarized the best evidence regarding the insertion and maintenance of built-in fecal incontinence management device. We recommend that clinical practitioners integrate this evidence into their practice, while considering individual patient preferences and medical contexts. Adhering to individualization for evidence translation improves standardization and benefits patients in the clinical use of fecal collection devices.

Objective:To explore the effect of specific inhibitor of Smad3 (SIS3) on glucocorticoid-induced ocular hypertension in mice and its possible mechanism.Methods:Fifty-one eight-week-old female C57BL/6J mice were randomly divided into control group, dexamethasone group and SIS3 group by the random number table method, with 17 mice in each group.Mice in the control group were injected with 20 μl 2 % polyvinyl alcohol into the conjunctival fornix every week for 4 weeks.Mice in the dexamethasone group and SIS3 group were injected with 20 μl 10 mg/ml dexamethasone acetate every week and SIS3 group was treated with additional 100 μg/ml SIS3 nanomicelle eye drops 3 times daily for up to 4 weeks.Intraocular pressure (IOP) was measured weekly using Icare rebound tonometer.Mice were sacrificed 4 weeks after treatment, and the eyeballs were removed.Morphology of trabecular meshwork (TM) tissues were detected by hematoxylin-eosin (HE) staining.The collagen deposition area in TM tissues were examined by Masson staining.Fibronectin (FN) and collagen type Ⅰ (Col-1) in the extracellular matrix of TM tissue were detected by immunofluorescence staining.TM tissues were obtained from donated patients, and primary human trabecular meshwork cells (HTMCs) were obtained by culture.The expression level of myocilin in dexamethasone-induced HTMCs was detected by immunofluorescence and Western blot for cell identification.Primary HTMCs were divided into normal control group, dexamethasone group and SIS3 group cultured with normal culture medium, medium containing 400 nmol/L dexamethasone, medium containing 400 nmol/L dexamethasone+ 10 μmol/L SIS3 for 48 hours, respectively.The expression levels of FN, Col-1 and p-Smad3/Smad3 proteins were measured by Western blot.The use and care of animals complied with the ARVO statement.This study protocol was approved by the Animal Ethics Committee of Zhengzhou University (No.ZZU-LA20220729).The collection of TM tissue specimens complied with the Declaration of Helsinki and was approved by the Medical Ethics Committee of Henan Provincial Eye Hospital (No.HNEECKY-2022[18]).The patients knew the purpose of the experiment and signed the informed consent forms.Results:There was a significant overall difference in IOP among the three groups at different time points after administration ( Fgroup=72.94, P<0.001; Ftime=33.19, P<0.001).Compared with baseline, IOP was increased in the dexamethasone group at each time point after administration, and the differences were statistically significant (all P<0.001).The IOP of the control and SIS3 groups at weeks 1, 2, 3, 4 were significantly lower than that of the dexamethasone group (all P<0.001).HE staining showed that the iridocorneal angles of all groups were open with similar morphology of the TM structure.Masson staining showed that the positive expression area of collagen in the control group, dexamethasone group and SIS3 group was (9.57±2.91)%, (27.75±5.88)% and (11.67±3.78)%, respectively, with a statistically significant difference among the three groups ( F=25.91, P<0.001), and the positive expression area of collagen was significantly lower in the control group and SIS3 group than in the dexamethasone group (all P<0.001).The fluorescence expression level of FN in the control group, dexamethasone group and SIS3 group was 8.00±1.92, 14.01±2.74 and 7.85±0.64, respectively, and the fluorescence expression level of Col-1 was 6.90±1.16, 14.36±3.19 and 4.90±0.88, respectively, with statistically significant differences among the three groups ( F=15.93, 30.29; both P<0.001), and the fluorescence expression levels of FN and Col-1 were significantly lower in the control group and SIS3 group than in the dexamethasone group (all P<0.01).Immunofluorescence staining and Western blot showed that the cultured primary cells expressed myocilin and the expression level of myocilin was significantly increased after dexamethasone induction, which was identified as HTMCs.There were statistically significant differences in the relative expression levels of FN, Col-1, and p-Smad3/Smad3 proteins among different groups of cells ( F=8.22, 23.08, 8.78; all P<0.05), and the relative expression levels of FN, Col-1, and p-Smad3/Smad3 proteins were significantly lower in the control group and SIS3 group than in the dexamethasone group (all P<0.05). Conclusions:SIS3 reduces IOP by inhibiting p-Smad3, reducing extracellular matrix deposition in TM, and reducing fibrosis in the TM tissue.

Approximately 60% of colorectal cancer (CRC) patients exhibit TP53 mutations, which are strongly associated with tumor progression, chemotherapy resistance, and an unfavorable prognosis. However, targeting p53 has historically been challenging, and currently, there are no approved p53-based therapeutics for clinical use worldwide. In this study, we discovered that ubiquitin carboxyl terminal hydrolase L3 (UCHL3) plays a crucial role in high-level glycolysis, enhanced stem-like properties, and 5-fluorouracil (5-FU) chemoresistance in TP53-mutant CRC by exerting its deubiquitinating enzyme activity to stabilize α-enolase (ENO1) protein. Notably, we identified a newly Food and Drug Administration (FDA)-approved drug, pacritinib, that potently suppresses UCHL3 expression by blocking the janus kinase 2 (JAK2)-signal transducer and activator of transcription 3 (STAT3) pathway in TP53-mutant CRC. Furthermore, Pacritinib was demonstrated to effectively inhibit glycolysis and improve the sensitivity to 5-FU chemotherapy in TP53-mutant CRC. Our findings suggest that targeting the JAK2-STAT3-UCHL3-ENO1 axis is a promising strategy to suppress glycolysis and enhance the efficacy of 5-FU chemotherapy in TP53-mutant CRC. Pacritinib shows potential for clinical application in the treatment of TP53-mutant CRC.

OBJECTIVE: To explore the clinical and genetic characteristics of two children diagnosed with two rare genetic diseases simultaneously. METHODS: Two children with comorbidity of two genetic diseases due to dual genetic mutations diagnosed at the Third Affiliated Hospital of Zhengzhou University respectively in May 2022 and March 2023 were selected as the study subjects. Clinical and genetic data of the two children were retrospectively analyzed. This study has been approved by the Ethics Committee of the Third Affiliated Hospital of Zhengzhou University (Ethic No. 2021-062-01). RESULTS: Child 1 was a 2-year-and-4-month-old boy whose clinical manifestations included facial dysmorphism, developmental delay, short stature, microcephaly, cleft palate, cryptorchidism, hypospadias, recurrent infections and immunological abnormalities. Whole exome sequencing revealed that he had harbored a heterozygous c.6595delT (p.Y2199Ifs*65) variant of the KMT2D gene and a heterozygous c.1892G>A (p.R631Q) variant of the PIK3R1 gene. This has led to a dual genetic diagnosis of Kabuki syndrome and PI3Kδ-related immunodeficiency type 36. Child 2 was a 15-year-old girl whose clinical manifestations included epilepsy, Albright's hereditary osteodystrophy, long body trunk, short limbs, hypocalcemia, hyperphosphatemia and hyperparathyroidism. The child also had a family history of short stature. Whole exome sequencing revealed that she had harbored a heterozygous c.2T>C (p.Met1?) variant of the GNAS gene and deletion of exons 2 to 6 of the SHOX gene. The two variants have led to dual diagnose of pseudohypoparathyroidism and X-linked idiopathic short stature. CONCLUSION When the clinical phenotype of a genetic disease is complex and cannot be fully explained with a single genetic variant, multiple pathogenic variants should be considered, and this may lead to the diagnosis of co-morbid genetic diseases. To adopt or supplement corresponding genetic testing in time and re-analyze the genetic data may facilitate accurate diagnosis of co-morbid genetic diseases.
Child, Preschool ; Female ; Humans ; Male ; Class Ia Phosphatidylinositol 3-Kinase/genetics* ; Comorbidity ; Exome Sequencing ; Mutation ; Rare Diseases/genetics* ; Retrospective Studies ; Adolescent

Objective:To explore the clinical and genetic characteristics of two children diagnosed with two rare genetic diseases simultaneously.Methods:Two children with comorbidity of two genetic diseases due to dual genetic mutations diagnosed at the Third Affiliated Hospital of Zhengzhou University respectively in May 2022 and March 2023 were selected as the study subjects. Clinical and genetic data of the two children were retrospectively analyzed. This study has been approved by the Ethics Committee of the Third Affiliated Hospital of Zhengzhou University (Ethic No. 2021-062-01).Results:Child 1 was a 2-year-and-4-month-old boy whose clinical manifestations included facial dysmorphism, developmental delay, short stature, microcephaly, cleft palate, cryptorchidism, hypospadias, recurrent infections and immunological abnormalities. Whole exome sequencing revealed that he had harbored a heterozygous c.6595delT (p.Y2199Ifs*65) variant of the KMT2D gene and a heterozygous c. 1892G>A (p.R631Q) variant of the PIK3R1 gene. This has led to a dual genetic diagnosis of Kabuki syndrome and PI3Kδ-related immunodeficiency type 36. Child 2 was a 15-year-old girl whose clinical manifestations included epilepsy, Albright′s hereditary osteodystrophy, long body trunk, short limbs, hypocalcemia, hyperphosphatemia and hyperparathyroidism. The child also had a family history of short stature. Whole exome sequencing revealed that she had harbored a heterozygous c. 2T>C (p.Met1? ) variant of the GNAS gene and deletion of exons 2 to 6 of the SHOX gene. The two variants have led to dual diagnose of pseudohypoparathyroidism and X-linked idiopathic short stature. Conclusion:When the clinical phenotype of a genetic disease is complex and cannot be fully explained with a single genetic variant, multiple pathogenic variants should be considered, and this may lead to the diagnosis of co-morbid genetic diseases. To adopt or supplement corresponding genetic testing in time and re-analyze the genetic data may facilitate accurate diagnosis of comorbid genetic diseases.

OBJECTIVE To observe the influence of carbapenem-resistant Klebsiella pneumoniae(CRKP)infec-tion/colonization on mortality risk of the intensive care unit(ICU)patients.METHODS The patients who were hospitalized in ICUs of West China Hospital of Sichuan University from Jan.1,2016 to Dec.31,2018 were recrui-ted as the research subjects.By means of retrospective cohort study,the enrolled patients were divided into the CRKP infection/colonization group and the non-CRKP infection/colonization group according to the status of isola-tion of CRKP strains from the clinical specimens of the ICU patients.The 30-day mortality risk of the CRKP in-fection/colonization group and the non-CRKP infection/colonization group was analyzed by Kaplan-Meier.The in-fluencing factors for the 30-day mortality risk of the ICU patients were analyzed by means of Cox proportional haz-ard model.RESULTS A total of 2229 patients were enrolled in the study,of which 89 were assigned as the CRKP infection/colonization group,and 2140 were assigned as the non-CRKP infection/colonization group.The sputum was the major specimen source from the patients with CRKP infection/colonization,and the lower respira-tory tract was the major infection site.The 30-day survival rate was 66.49％in the CRKP infection/colonization group,78.49％in the non-CRKP infection/colonization group,and there was significant difference(x2=7.200,P=0.007).The Cox proportional hazard model analysis showed that the CRKP infection/colonization could in-crease the 30-day mortality risk of the ICU patients(HR=1.839,95％CI:1.126 to 3.002,P=0.015);the age(HR=1.014,95％CI:1.006 to 1.022,P＜0.001),APACHE Ⅱ(HR=1.035,95％CI:1.018 to 1.053,P＜0.001),use of caspofungin(HR=1.398,95％CI:1.038 to 1.882,P=0.028),central venous catheter indwelling(HR=3.752,95％CI:1.808 to 7.790,P＜0.001)and blood purification(HR=2.061,95％CI:1.518 to 2.797,P＜0.001)may also increase the 30-day mortality risk of the ICU patients.CONCLUSIONS The CRKP infection/colonization patients are at higher 30-day mortality risk than the non-CRKP infection/colonization patients.It is necessary to formulate and implement the prevention and control measures to reduce the incidence of CRKP infection/colonization and take measures to reduce the mortality rate of the patients so as to improve the prognosis.

OBJECTIVE To observe the influence of carbapenem-resistant Klebsiella pneumoniae(CRKP)infec-tion/colonization on mortality risk of the intensive care unit(ICU)patients.METHODS The patients who were hospitalized in ICUs of West China Hospital of Sichuan University from Jan.1,2016 to Dec.31,2018 were recrui-ted as the research subjects.By means of retrospective cohort study,the enrolled patients were divided into the CRKP infection/colonization group and the non-CRKP infection/colonization group according to the status of isola-tion of CRKP strains from the clinical specimens of the ICU patients.The 30-day mortality risk of the CRKP in-fection/colonization group and the non-CRKP infection/colonization group was analyzed by Kaplan-Meier.The in-fluencing factors for the 30-day mortality risk of the ICU patients were analyzed by means of Cox proportional haz-ard model.RESULTS A total of 2229 patients were enrolled in the study,of which 89 were assigned as the CRKP infection/colonization group,and 2140 were assigned as the non-CRKP infection/colonization group.The sputum was the major specimen source from the patients with CRKP infection/colonization,and the lower respira-tory tract was the major infection site.The 30-day survival rate was 66.49％in the CRKP infection/colonization group,78.49％in the non-CRKP infection/colonization group,and there was significant difference(x2=7.200,P=0.007).The Cox proportional hazard model analysis showed that the CRKP infection/colonization could in-crease the 30-day mortality risk of the ICU patients(HR=1.839,95％CI:1.126 to 3.002,P=0.015);the age(HR=1.014,95％CI:1.006 to 1.022,P＜0.001),APACHE Ⅱ(HR=1.035,95％CI:1.018 to 1.053,P＜0.001),use of caspofungin(HR=1.398,95％CI:1.038 to 1.882,P=0.028),central venous catheter indwelling(HR=3.752,95％CI:1.808 to 7.790,P＜0.001)and blood purification(HR=2.061,95％CI:1.518 to 2.797,P＜0.001)may also increase the 30-day mortality risk of the ICU patients.CONCLUSIONS The CRKP infection/colonization patients are at higher 30-day mortality risk than the non-CRKP infection/colonization patients.It is necessary to formulate and implement the prevention and control measures to reduce the incidence of CRKP infection/colonization and take measures to reduce the mortality rate of the patients so as to improve the prognosis.

Objective:To search, evaluate, and summarize the best evidence of built-in fecal incontinence management device, to inform the management of incontinence dermatitis by clinical healthcare professionals.Methods:BMJ Best Practice, UpToDate, Guideline International Network, Joanna Briggs Institute, National Institute for Health and Care Excellence, Scottish Intercollegiate Guidelines Network, Registered Nurses′Association of Ontario, The Cochrane Library, Medline, Embase, SinoMed, CINAHL, PubMed, Web of Science, OVID, China National Knowledge Infrastructure, Wanfang Database were systematically searched for all evidence regarding the application of fecal collection devices. It included clinical practice, guidelines, systematic reviews, expert consensuses, evidence summaries, and randomized controlled trial. Two researchers independently evaluated the literature quality and extracted the literature that met the standards.Results:A total of 12 pieces of the literature were involved, including 2 best practice, 5 guidelines, 3 expert consensuses, and 2 systematic reviews. This study summarized 26 pieces of best evidence in relation to the following 5 themes: indications and contraindications, device insertion, device maintenance, device removal and effectiveness evaluation.Conclusions:This study scientifically and systematically summarized the best evidence regarding the insertion and maintenance of built-in fecal incontinence management device. We recommend that clinical practitioners integrate this evidence into their practice, while considering individual patient preferences and medical contexts. Adhering to individualization for evidence translation improves standardization and benefits patients in the clinical use of fecal collection devices.

Objective:To explore the clinical and genetic characteristics of two children diagnosed with two rare genetic diseases simultaneously.Methods:Two children with comorbidity of two genetic diseases due to dual genetic mutations diagnosed at the Third Affiliated Hospital of Zhengzhou University respectively in May 2022 and March 2023 were selected as the study subjects. Clinical and genetic data of the two children were retrospectively analyzed. This study has been approved by the Ethics Committee of the Third Affiliated Hospital of Zhengzhou University (Ethic No. 2021-062-01).Results:Child 1 was a 2-year-and-4-month-old boy whose clinical manifestations included facial dysmorphism, developmental delay, short stature, microcephaly, cleft palate, cryptorchidism, hypospadias, recurrent infections and immunological abnormalities. Whole exome sequencing revealed that he had harbored a heterozygous c.6595delT (p.Y2199Ifs*65) variant of the KMT2D gene and a heterozygous c. 1892G>A (p.R631Q) variant of the PIK3R1 gene. This has led to a dual genetic diagnosis of Kabuki syndrome and PI3Kδ-related immunodeficiency type 36. Child 2 was a 15-year-old girl whose clinical manifestations included epilepsy, Albright′s hereditary osteodystrophy, long body trunk, short limbs, hypocalcemia, hyperphosphatemia and hyperparathyroidism. The child also had a family history of short stature. Whole exome sequencing revealed that she had harbored a heterozygous c. 2T>C (p.Met1? ) variant of the GNAS gene and deletion of exons 2 to 6 of the SHOX gene. The two variants have led to dual diagnose of pseudohypoparathyroidism and X-linked idiopathic short stature. Conclusion:When the clinical phenotype of a genetic disease is complex and cannot be fully explained with a single genetic variant, multiple pathogenic variants should be considered, and this may lead to the diagnosis of co-morbid genetic diseases. To adopt or supplement corresponding genetic testing in time and re-analyze the genetic data may facilitate accurate diagnosis of comorbid genetic diseases.