[The mechanism of OC-STAMP overexpression induced actin cytoskeleton remodeling in promoting epithelial-mesenchymal transition in the alveolar type â ¡ epithelial cell].

Objective: To explore the mechanism of osteoclast stimulatory transmembrane protein (OC-STAMP) overexpression on epithelial-mesenchymal transition (EMT) . Methods: In April 2021, mice alveolar type Ⅱ epithelial cells MLE-12 were divided into five groups: overexpression control group (NC group), Ocstamp overexpression group (over-Ocstamp group), Fasudil intervention group (over-Ocstamp+Fasudil group), silence control group (si-NC group), Ocstamp silence group (si-Ocstamp group). The protein expressions of OC-STAMP, epithelial marker protein-E-cadherin (E-cad), interstitial marker protein-α-smooth muscle actin (α-SMA), Ras homolog gene family member A (RhoA), Rho GDP dissociation inhibitor α (Rho GDIα), Rho-associated protein kinase (ROCK), phosphate myosin phosphatase (p-MYPT) were examined by Western blotting and Immunocytochemical staining. The filamentous actin (F-actin) was detected by Phalloidin method. t test was used to compare the relative expression of each protein between the two groups. Results: Western blotting and Immunocytochemical staining showed that compared with the NC group, the expression level of E-cad was down-regulated, while the expression levels of α-SMA, Rho GDIα, RhoA, ROCK, p-MYPT were increased, and F-actin expression was enhanced in the over-Ocstamp group. The differences were statistically significant (P<0.05). There were no significant differences in E-cad and α-SMA protein expression in si-Ocstamp group compared with si-NC group (P>0.05). Compared with over-Ocstamp group, the expression level of E-cad protein in over-Ocstamp+Fasudil group was up-regulated, the expression levels of α-SMA, Rho GDIα, RhoA, ROCK and p-MYPT protein were decreased, and F-actin expression was weakened, with statistical significance (P<0.05) . Conclusion: OC-STAMP overexpression in alveolar type Ⅱ epithelial cells may induce actin cytoskeleton remodeling through activation of Rho GDIα/RhoA/ROCK signaling pathway, thus promoting EMT.

[Characteristics of SARS-CoV-2 Omicron infection in children imported from Hong Kong].

Objective: To understand the clinical characteristics of children with SARS-CoV-2 Omicron infection imported from Hong Kong Special Administrative Region. Methods: This retrospective study was conducted to collect the data including clinical manifestations, outcomes and vaccination of 107 children with SARS-CoV-2 Omicron infection imported from Hong Kong Special Administrative Region to be admitted to the designated referral hospital in Shanghai from February to March 2022. According to the occurrence of clinical symptoms, the cases were divided into asymptomatic group and symptomatic group. According to the age of diagnosis, the cases were divided into <3 years group, 3-<6 years group and 6-<18 years group, and the clinical manifestations in different age group were analyzed with t-test and Mann-Whitney rank-sum test. Besides, to analyze the effectiveness of vaccination against SARS-CoV-2 Omicron infection in different age group, the cases aged 3-<18 years were also subdivided into unvaccinated group, 1-dose group and 2-dose group, and the relative risk (RR) was used to demonstrate the effectiveness. Results: Among the 107 cases, 66 were male and 41 were female, with infection age of 10 (5, 14) years. There were 29 cases in the asymptomatic group, and 78 cases in the symptomatic group, and no significant difference in the age of infection was observed between the 2 groups (11 (6, 14) vs. 10 (5, 14) years, Z=0.49, P>0.05). And there were no severe cases in symptomatic group. The length of hospitalization was (18±6) days, and was longer in symptomatic group than that in asymptomatic group ((19±6) vs. (16±7) d, t=0.17, P=0.030). Eight-two cases (76.6%) had a history of epidemiological exposure and, among whom, 81 cases (75.7%) were associated with household transmission. Among symptomatic group, 57 cases (73.1%) had fever and 20 cases (25.6%) had cough. Of the 74 cases undergoing chest CT examination, 17 cases (23.0%) showed mild abnormalities. Of the 83 cases who received the lab tests, 23 cases (27.7%) had white blood cell counts<4×109/L, 3 cases (3.6%) had C-reaction protein >8.0 mg/L, and 6 cases (7.2%) had slightly elevated aspartate transaminase and alanine aminotransferase. Among the 92 children aged 3-<18 years, 31 cases were unvaccinated, 34 cases received 1 dose, and 27 cases received 2 doses. The interval between the last vaccination and infection was 2.2 (0.6, 6.0) months; the interval between the last vaccination and infection in the 2-dose group was longer than that in 1-dose group (6.0 (4.5, 7.3) vs. 0.7 (0.3,2.0) months, Z=3.59, P<0.001).The risk of symptomatic infection was reduced by 45% (RR=0.55, 95% CI 0.35-0.87) with two-dose vaccination compared to non-vaccination in cases aged 3-<18 years. All these cases recovered completely. Conclusions: Children infected with SARS-CoV-2 Omicron are usually mild or asymptomatic. Household transmission is the main pattern of infection with SARS-CoV-2 Omicron in children. Two-dose SARS-CoV-2 vaccination in children aged 3-<18 years can provide partial protection against disease caused by SARS-CoV-2 Omicron.

[Clinical analysis of liver chemistries in children and adolescent with 2019-nCoV infection].

Objective: To describe the clinical features of liver involvement in children and adolescent with 2019-nCoV infection. Methods: The clinical data of 77 hospitalized cases admitted to the Children's Hospital of Fudan University were collected from January 19 to November 28, 2020. The characteristics and risk factors of abnormal liver chemistries in children with laboratory-confirmed 2019-nCoV infection were analyzed. Results: Of the 77 cases, 44 were male (57.1%) and 33 were female (42.9%), with a median age of 10 years. 27(35.1%) were asymptomatic, 28(36.4%) had mild illness, 22(28.6%)had non-severe pneumonia. Hydroxychloroquine was used in 7 cases. Of the 75 children without underlying diseases, alanine aminotransferase was elevated in 1 case (1.5%, during hydroxychloroquine therapy), aspartate aminotransferase was elevated in 7 cases (10.3%), alkaline phosphatase was elevated in 7 cases (28%), and total bilirubin, direct bilirubin, albumin, international normalized ratio were in normal range. There was no statistical difference between the pneumonia group and the non-pneumonia group in term of liver chemistries (P > 0.05), same as between the elevated erythrocyte sedimentation rate group and the normal group. There was no aggravation of liver injury in the child with biliary atresia. The child with epilepsy showed no abnormal liver chemistries after infection. Conclusion: Children with 2019-nCoV infection had mild clinical symptoms with few cases of liver injury. The abnormal liver chemistries in children with COVID-19 infection may be related to the underlying disease and the use of antiviral drugs.

Downregulation of nuclear protein-1 induces cell cycle arrest in G0/G1 phase in glioma cells in vivo and in vitro via P27.

Nuclear protein-1 (NUPR1), also named as p8 or Com1, has been since found overexpressed in several human malignant tumor cells, such as glioma. NUPR1 also regulates cell cycle progression, however, the role of NUPR1 in regulating glioma cell cycle remains poorly understood. Knockdown efficiency of U87 and U251 cells infected with the lentiviral vector was detected by quantitative real-time PCR and western blot in vitro and in vivo. Flow cytometry and western blot were used to explore a mechanism by which NUPR1 modulates cell cycle in U87 and U251 cells. Immunohistochemistry was applied to detect expression levels of P27, CDK2, and cyclin E in human glioma tissues with NUPR1 positive expression and tumorigenesis in nude mice. We confirmed that the downregulation of NUPR1 arrested the cell cycle in the G0/G1 phase in U87 and U251 cells in vitro. Furthermore, the expression level of P27 was increased, and CDK2 and cyclin E were decreased upon silencing NUPR1 expression in vitro and in vivo. In conclusion, the knockdown of NUPR1 induces cell cycle arrest in the G0/G1 phase in glioma cells via P27.

[Sacral neuromodulation preliminary outcomes in male patients with idiopathic dysuria].

Objective: To assess the effectiveness and safety of sacral neuromodulation (SNM) therapy for men with idiopathic dysuria. Methods: From January 2012 to December 2016, a total of 26 patients treated with SNM therapy from multi-center across the country were retrospectively studied. The age ranged from 19 to 86 years with an average age of 45.2 years. Patients suffered from one or multiple urinary symptoms such as frequency of urination, urgency, urinary retention, etc. All patients had received more than two types of conservative therapy including oral and behavioral therapy, but had poor or no improvement. The voiding diary, urgency score and the quality of life score before implantation, in stageⅠ after implantation and stage Ⅱ after permanent implantation were recorded and compared. Results: A total of 22 patients chose to receive IPG at the end of stageⅠ therapy while 4 patients refused further stage Ⅱ therapy because of dissatisfactory effect. The conversion rate of stage Ⅰ to stage Ⅱ was 84.6% (22/26). The average follow-up time was 19.2 months, ranging from 3 to 63 months. The baseline of residual urine, voiding frequency and average voiding amount and those after stage Ⅰ therapy were [5 (0, 137.5) ] ml vs [0 (0, 40) ] ml, 14.6±6.1 vs 9.1±2.8, [100 (80, 135) ] ml vs [190 (150, 210) ] ml, respectively. The differences were statistically significant(all P<0.05).However, no significant differences were found in urination volume and quality of life (QoL) before and after implantation(P>0.05). During an average follow-up time of 19.2 months after the permanent implantation, over 80% patients had an improvement of residual urine volume by more than 50% after permanent implantation while the improvement was 55.6% after stage Ⅰ therapy, suggesting that the improvement of residual urine volume might be positively correlated with the duration of regulation. No significant differences were found in other parameters between stage Ⅰ and Ⅱ therapy. No adverse events like wound infection and electrode dislocation happened during our study. Conclusions: SNM is an effective and safety procedure for male patients with idiopathic dysuria, with a relatively high transfer rate. The medium-term curative effect is stable. The duration of regulation may be positively correlated with the improvement of residual urine.

[Risk factors associated with systemic inflammatory response syndrome after flexible ueteroscopic lithotripsy based on enhanced recovery after surgery].

Objective: To investigate the risk factors of systemic inflammatory response syndrome (SIRS) in patients undergoing flexible ureteroscopic lithotripsy based on enhanced recovery after surgery (ERAS). Methods: The clinical data of 243 kidney stone cases who underwent flexible ureteroscopic lithotripsy based on ERAS in the First Affiliated Hospital of Wannan Medical College from January 2016 to December 2017 were analyzed retrospectively. The cases were divided into two groups according to whether they had SIRS after surgery: SIRS group (26 cases) and non-SIRS group (217 cases). The age, gender, laterality of kidney stone, history of previous kidney stone surgery, degree of hydronephrosis, multiple kidney stones, length of operation time, white blood cell count of preoperative urine routine, result of preoperative urine culture, use of preoperative antibiotics, diabetes and other chronic diseases in the groups were collected and analyzed. Results: SIRS occurred in 26 cases in this study, which accounted for 10.7% (26/243). Multivariate analysis found that, moderate and severe hydronephrosis (OR=6.711, P=0.008), stone burden ≥2 cm (OR=10.353, P<0.001), length of operation time ≥ 60 min (OR=5.583, P=0.011), white blood cell count of preoperative urine routine ≥25×10(6)/L (OR=6.195, P=0.005), positive preoperative urine culture (OR=4.216, P=0.011), diabetes and other chronic diseases (OR=4.532, P=0.006) were the independent risk factors for postoperative SIRS (P<0.05). Conclusions: The occurrence of SIRS after flexible ureteroscopic lithotripsy based on ERAS is closely correlated with hydronephrosis, stone burden, length of operation time, white blood cell count of preoperative urine routine, positive preoperative urine culture, diabetes and other chronic diseases.

[Clinical epidemiology and antimicrobial resistance of nontyphoidal Salmonella enteric infections in children: 2012-2014].

OBJECTIVE: To understand the clinical epidemiology and manifestation of nontyphoidal Salmonella (NTS)enteric infections and serotypes and antibiotic resistance patterns of NTS in children. METHOD: All cases with probable acute bacterial inflammatory diarrhea were included into this study from the Children's Hospital of Fudan University during January 2012 through December 2014. Salmonella species and other major enteric bacteria were routinely isolated from the fresh stool sample of the enrolled patients at their first visit to the enteric clinic. Salmonella isolates were serotyped and tested for antimicrobial susceptibility testing using Kirby-Bauer method at the reference laboratory of Shanghai Municipal Center for Disease Control and Prevention. All enrolled cases had clinical records. The enumeration data were described by constituent ratio. RESULT: Of the enrolled 3 143 patients, 742 (23.6%) were confirmed to have Salmonella infections, and NTS was identified in 738 (23.5%) cases and Salmonella Paratyphi B was identified in 4 (0.1%) cases, respectively. The isolation rate of NTS exceeded diarrheagenic Escherichia coli (6.7%, 182/2 727), Campylobacter (4.1%, 129/3 143) and Shigella (2.4%, 77/3 143). Nine serogroups and 41 serovars were identified in 742 salmonella isolates. Salmonella Enteritidis and Salmonella Typhimurium were the most common serovars, representing 42.6% (316/742) and 28.2% (209/742) of isolates, respectively.NTS infections were prevalent year-round with a seasonal peak from May to October. The common symptoms included fever (57.5%, 424/738) and blood-in-stool (31.0%, 229/738). Antimicrobial susceptibility showed only 8 (1.1%) of 742 isolates sensitive to 16 tested antibiotics.Resistance to the current first-line antibiotics such as the third-generation and(or) fourth-generation cephalosporins (cefotaxime and cefepime), ciprofloxacin was detected in 10.0%(74/738), 8.1%(60/738) and 1.1%(8/738), respectively. CONCLUSION: NTS is the leading causative agent responsible for bacterial inflammatory diarrhea in Shanghainese children. The prevalence of NTS enteric infections show an increasing trend year by year. It is essential to continuously monitor the antimicrobial resistance of NTS and trace the source of infection to prevent and treat NTS infections in Shanghai.

Alleviation of pre-exposure to low-dose 16O8+ ion on mouse testicular histological damage induced by subsequent high-dose irradiation.

The testes of the B6C3F1 hybrid strain mice were irradiated with 0.05 Gy of 16O8+ ion as the pre-exposure dose (D1), and were then irradiated with 2 Gy of 16O8+ ion as challenging radiation dose (D2) at 4 h after per-exposure. Testicular morphology was observed by light microscope at 35th day after radiation. The results showed that irradiation of mouse testes with 2 Gy of 16O8+ ion significantly impaired, mainly reduction of tubule diameter and decrease or loss of germ cells in various developing stages, especially spermatogenic elements. Pre-exposure to a low-dose (0.05 Gy) of 16O8+ ion significantly alleviated above mentioned damage on testicular morphology induced by subsequent a high-dose (2 Gy) radiation.

Effects of 16O+6 ion irradiation on human sperm spontaneous chemiluminescence, motility, acrosome reaction and viability in vitro.

Effects of 16O+6 ion irradiation with different doses on human sperm spontaneous chemiluminescence (SCL), motility, acrosome reaction (AR) and viability were examined. Spermatozoa were irradiated with 0, 0.25, 0.5, 1, 2, 4, 8, 16, 32, or 64 Gy 16O+6 ion beam at the energy of 3.17 MeV/u. After irradiation, samples were analyzed by SCL measurement at 1, 2 and 3 h of incubation; motility was determined by the transmembrane migration method within 2 h of incubation; the percentage of AR and viability was evaluated by the triple-stain technique at 3.5 h of incubation. The results showed: sperm SCL was significantly increased with irradiation doses and the lowest effective dose was 0.5 Gy; compared with controls, the transmembrane migration ratio of spermatozoa progressively elevated with irradiation doses at 0.5, 1, and 2 Gy; the percentage of sperm AR markedly increased in 0.5-4 Gy irradiation and the optimal dose was 2 Gy, and then significant decreased with further increase of irradiation doses; the viability had no significant change within 0.25-8 Gy, but was progressively decreased at 16, 32 and 64 Gy. These data suggested that heavy ion at low doses increased motility and AR, whereas had deleterious effects at higher doses, which are associated with free radical reactions induced by heavy ion irradiation.

Chromosomal aberrations induced by 12C6+ heavy ion irradiation in spermatogonia and spermatocytes of mice.

The testes of Kun-Ming strain mice were radiated with different doses of 12C6+ ion or 60Co gamma-ray. Chromosomal aberrations induced in spermatogonia and spermatocytes were analyzed by the air-drying method. The relative biological effectiveness (RBE) of 12C6+ ion was calculated with respect to 60Co gamma-ray for the induction of chromosomal aberrations. The 12C6+ ion and 60Co gamma-ray dose-response relationships for chromosomal aberrations were plotted by linear quadratic models. The results showed that there was an increase in frequency of chromosomal aberrations in all the treated groups compared to controls. The RBE values were 1.67 for aberrations of spermatogonia and 1.66 for aberrations of spermatocytes for a dose of 2.0 Gy. Moreover, a different distribution of the various types of aberrations has been found for 12C6+ ion and 60Co gamma-ray irradiations. The dose-response relationships for 12C6+ ion and 60Co gamma-ray exhibited negative curvature in both spermatogonia and spermatocytes groups: the frequencies of aberrations increased sharply at low doses and exhibited less sharp increases for higher doses, which may be related to an interaction between the chromosomal damage and a block in cell cycle. Our results may provide useful information for the assessment of genetic risks of humans exposed to heavy ions.

Effects of pre-exposure of mouse testis with low-dose (16)O8+ ions or 60Co gamma-rays on sperm shape abnormalities, lipid peroxidation and superoxide dismutase (SOD) activity induced by subsequent high-dose irradiation.

PURPOSE: To investigate the effects of pre-exposure of mouse testis with low-doses of (16)O8+ ions or 60Co gamma-rays on sperm shape abnormalities, lipid peroxidation and superoxide dismutase (SOD) activity induced by subsequent high-dose irradiation. MATERIALS AND METHODS: Testes of the B6C3F1 hybrid strain mice were pre-irradiated with 0.05 Gy of (16)O8+ ions or 60Co gamma-rays and then after 4 h given a test irradiation with 2 Gy of the same radiation type. SOD activity and thiobarbituric acid reactive substances (TBARS) in the testes were determined by spectrophotometric and TBA methods respectively at 4 h after irradiation. Testis weight, sperm count and sperm morphology were analysed at day 35 after irradiation. RESULTS: Compared with controls, there was a significant increase in SOD activity and a significant decrease in TBARS level of pretreated testes. Testis weight loss, sperm count reduction and sperm abnormalities were significantly lower in the pretreated testes. The bioeffects of a 2 Gy dose of (16)O8+ ions relative to 60Co gamma-rays were 1.84 +/- 0.28 for testis weight, 1.22 +/- 0.25 for sperm count and 1.29 +/- 0.10 for sperm abnormalities. CONCLUSIONS: These data suggest that pre-exposure of testes with a low dose of heavy ions or gamma-rays renders the organ more resistant to subsequent high-dose irradiation. The increase of SOD activity and the decrease of lipid peroxidation levels induced by low-dose ionizing irradiation may be involved in this resistance. The effects with heavy ion irradiation were greater than with gamma-rays.