Objective:To evaluate the effect of nicastrin (nct) gene on the biological functions of melanocytes in zebrafish.Methods:By using a morpholino oligonucleotide (MO) technology, a nct-MO sequence targeting the zebrafish nct mRNA was designed, so was a MO control (ctrl-MO) sequence. Then, the enhanced green fluorescent protein (EGFP) mRNA with MO target sequence at its 5′ end was synthesized, and co-microinjected with the nct-MO or ctrl-MO sequence into the zebrafish embryos to verify the silencing efficiency of nct-MO and observe changes in developmental phenotypes in zebrafish. With wild-type zebrafish as a blank control group, real-time fluorescence-based quantitative PCR (RT-PCR) was conducted to determine the mRNA expression of melanin synthesis-and notch signaling pathway-related genes, including mitfa, tyr, tyrp1a, tyrp1b, dct, pmela, notch1a, notch1b and hey1 genes. One-way analysis of variance was used for the comparison of means among multiple groups, and least significant difference (LSD) - t test for multiple comparisons. Results:Eight hours after zebrafish fertilization, green fluorescence was observed in the zebrafish embryos in the ctrl-MO+EGFP mRNA group, but not in the nct-MO+EGFP mRNA group or blank control group. Forty-eight hours after fertilization, the proportion of pigmented area among the whole area of the tail of zebrafish larvae was significantly lower in the nct-MO group (0.169 ± 0.083) than in the ctrl-MO group (0.258 ± 0.042, t=3.202, P=0.005) , and disorderly pigment distribution in the tails was observed in the nct-MO group. RT-PCR revealed significant differences in the mRNA expression of pmela, tyrp1a and hey1 genes among the nct-MO group, ctrl-MO group and blank control group (all P < 0.05) , but no significant difference was observed in the mRNA expression of mitfa, tyr, tyrp1b, dct, notch1a or notch1b genes among the 3 groups (all P>0.05) ; the relative expression levels of pmela and tyrp1a mRNAs were significantly lower in the nct-MO group (0.708 ± 0.028, 0.558 ± 0.136, respectively) than in the ctrl-MO group (1.023 ± 0.142, 1.016 ± 0.134, respectively, both P < 0.05) . Conclusion:The nct gene may affect biological functions of melanocytes by regulating melanin synthesis in zebrafish.

ObjectiveTo investigate the effects of nasal continuous positive airway pressure (nCPAP) and intubation in very low birth weight preterm infants. Methods One hundred and twenty-three very low birth weight preterm infants with respiratory distress within 60 minutes after birth were randomly assigned to nCPAP (n=63) or intubation group (n=60).Outcomes at 7,28 days and 36 corrected gestational weeks were assessed with x2 or t-test. ResultsThere were no significant difference in fatality rate and incidence of bronchopulmonary dysplasia between nCPAP group and intubation group [7.9％ (5/63) vs 6.6％(4/60),4.8％(3/63) vs 3.3％(2/60),x2 =0.07and 0.16,P＞0.05].In nCPAP group,the use of pulmonary sulfactant was 27.0％ (17/63),lower than that (83.3 ％,50/60) in intubation group (x2 =39.34,OR=0.3,90 ％ CI:0.2-0.6,P＜0.05) ;The nCPAP group had fewer ventilation support in 28 days [17.5％ (11/63) vs 25.0％ (15/60),OR=0.7,90％ CI:0.4-1.4] and 36 weeks [6.3％ (4/63) vs 8.3％ (5/60),OR=0.8,90％ CI:0.2-2.4] than those in intubation group but without statistical difference (x2=1.05 and 0.01,P＞0.05,respectively).The incidence of air leak in nCPAP group were lower than intubation group [11.1％ (7/63) vs 33.3％ (20/60),x2 =8.86,OR=0.3,90％00 CI:0.2-0.7,P＜0.05].There was no significant difference for other complications between two groups. ConclusionsIn very low birth weight preterm infants,early nCPAP dose not significantly reduce the fatality rate and the incidence of bronchopulmonary dysplasia as compared with intubation ventilation,but shorten the time of ventilation and lower the incidence of air leak.

Objective To carry out a comparative study between the Chinese 2008 and'92 staging system of nasopharyngeal carcinoma (NPC). Methods A total of 777 patients presented with untreated nondisseminated NPC who had received MRI scan of nasopharynx and neck were studied retrospectively. The clinical materials and information of imaging were collected. All patients were restaged according to the Chinese 2008 and 92 staging system of nasopharyngeal carcinoma. Distribution of T, N stage, survival and prognostic value were compared. 513 patients of the 777 cases were treated with conventional radiotherapy,264 cases with intensity modulated radiation therapy. Results The 3-year follow-up rate was 97.6%. The consistency of T stages was 95.0%. T, N and clinical stage distributions in two systems were similar ( Kappa = 0. 93, P = 0. 000; Kappa = 0. 58, P = 0. 000; Kappa = 0. 74, P = 0. 000). Local failure-free survival and disease specific survival were also similar. There was no difference of distant metastasis between N0 and N1(x2 = 1.94,P=0. 164), and a marginal difference between N1 and N2(x2 =3.83,P=0.051) in the Chinese'92 staging system. However, although there was also no difference of distant metastasis-free survival between No and N1a(x2 =0. 07,P =0. 797), ) the difference of overall survival among N1b, N2, and N3 were significant ( x2 = 4. 95, P = 0. 026; x2 = 6. 74, P = 0. 009) in the Chinese 2008 staging system. Conventional radiotherapy or intensity modulated radiation therapy was not a prognostic factor for survival ( x2 = 3.60,P =0. 058). It is reasonable for the Chinese 2008 staging system integrated lymph node characteristics such as laterality, level and extranodal neoplastic spread into the N staging criteria ( x2 = 6. 59, P = 0. 010; x2 =4.78,P=0. 029;x2=9.32,P=0. 002). Conclusions For the Chinese 2008 staging system, it was reasonable to simplify the previous T stage. The N stage showed a better predictive role of distant failure.The role of retropharyngeal lymph node in stage system needs further investigation.

Objective To compare the efficacy and safety of the non-invasive high frequency ventilation (NHFV) and non-invasive intermittent positive pressure ventilation (NIPPV)in neonates with respiratory distress syndrome (RDS) after extubation.Method Neonates with RDS from January 2015 to January 2016,who required high frequency ventilation after birth and were extubated after treatment were retrospectively studied.The enrolled patients were divided into NHFV group and NIPPV group to compare the rate of successful extubation within 7 days,non-invasive respiratory ventilation support time and complication incidence.Result In total 42 neonates were included,NHFV group were 21 cases and NIPPV group were 21 cases.The rates of successful extubation were not statistically different (71.4％ vs.80.9％,P ＞ 0.05);Compared with NIPPV group,NHFV group had shorter ventilation time [3.5 (2.2,4.1) d vs.4.6 (2.8,5.3)];the incidence of bronchopulmonary dysplasia,pneumothorax,intraventricular hemorrhage,periventricular leukomalacia,retinopathy of premature and necrotizing enterocolitis between groups were not statistically different (P ＞ 0.05).Conclusion NHFV is a new safe and efficient ventilation support method for extubated neonates,and needs further research.

Objective To analyze the epidemud growth factor receptor(EGFR)mutations in NSCLC patients in Fujian province.Methods Fresh specimens of lung cancer and corresponding normal lung tissue were collected from 50 cases of NSCLC patients.After DNA extraction,nested polymerase chain reaction (nested PCR)and direct deoxyribonucleic acid(DNA)sequencing were used to analyze EGFR gene mutations in NSCLC patients.Results EGFR mutations in tumors were identified from 13 of 50(26%)patients,including 10 cages of in-frame deletion in exon 19 and 3 cases of amino acid substitution in exon 21.Conclusion The mjor type of EGFR mutation in NSCLC patients in Fujian is in-frame deletion in exon 19.

Objective To explore the expressions and significances of the tumor stem cell markers CD133-2,CD24 and CD44s in head and neck squamous cell carcinoma (HNSCC) tissues and their association with the clinical pathologic characteristics.Methods Expressions of CD133-2,CD24 and CD44s were analyzed by immunohistochemistry (SP) in 83 cases of primary HNSCC tissues and 46 cases of normal epithelia.Clinicopathological indexes were assessed.Results In primary HNSCC tissues and normal epithelia,the expression rates of CD133-2 and CD24 were 9.64 ％ (8/83),21.74 ％ (10/46) and 90.36 ％ (75/83),46.67 ％ (21/46)respectively,which had statistically significances (x2 =15.040,5.818,P ＜ 0.05).CD44s expression was detected in primary HNSCC tissues and normal epithelia,but their staining scores had statistical significance (Z =-4.262,P ＜ 0.05).In primary HNSCC tissues,the expression of CD133-2 had negative correlation with differentiation degree (x2 =7.246,P ＜ 0.05),but CD24 and CD44s had positive correlation with differentiation degree (x2 =9.005,44.765,P ＜ 0.05).In addition,the expression of CD44s in primary HNSCC tissues had negative correlation with T classification (x2 =4.650,P ＜ 0.05).Conclusion The expressions of CD24 and CD44s in primary HNSCC tissues are highly up-regulated with tumor cells differentiation,and further research needs to be performed to discover whether or not CD24 and CD44s could be the markers of tumor stem cells of HNSCC.The expression of CD133-2 in primary HNSCC tissues is highly down-regulated with tumor cell differentiation.As one of the tumor stem cell markers of HNSCC,CD133-2 may play an important role in the development and clinical outcomes of tumor.

Objective To evaluate the effect of mild hypothermia on the expression of phosphorylated eukaryotic translation initiation factor 2α (p-eIf2αt) in the hippocampus in a mouse model of cerebral ischemia-reperfusion (I/R).Methods Sixty pathogen-free healthy male C57BL6 mice,aged 8-12 weeks,weighing 20-30 g,were divided into 3 groups (n =20 each) using a random number table:sham operation group (group S),group I/R and mild hypothermia group (group H).Cerebral I/R was induced by 15-min occlusion of bilateral common carotid arteries followed by reperfusion in chloral hydrate-anesthetized mice.Surface cooling was started immediately after reperfusion to maintain the rectal temperature at 32-34 ℃ for 3 h in group H.The neurologic deficit score was evaluated at 24 h of reperfusion.The mice were then sacrificed,brains were immediately removed,and hippocampi were isolated for examination of pathologic changes of hippocampal CA1 region and for determination of neuroapoptosis (by TUNEL) and expression of peIf2α (by Western blot).The apoptosis rate was calculated.Results Compared with group S,the neurologic deficit scores at 24 h of reperfusion and apoptosis rate of hippocampal neurons were significantly increased,and the expression of p-eIf2α was up-regulated in I/R and H groups (P＜0.05).Compared with group I/R,the neurologic deficit scores at 24 h of reperfusion and apoptosis rate of hippocampal neurons were significantly decreased,and the expression of p-eIf2α was down-regulated in group H (P＜ 0.05).Conclusion Mild hypothermia reduces endoplasmic reticulum stress and inhibits neuroapoptosis through inhibiting the expression of p-eIf2α in the hippocampus in a mouse model of cerebral I/R.

OBJECTIVE To evaluate the efficacy of otoendoscopic stapes surgery.METHODS From December 2020 to May 2023,a retrospective analysis was performed on 29 patients who underwent otoendoscopic stapes fenestration and artificial stapes implantation were diagnosed with otosclerosis during the operation.The medical history data of the patients were collected and analyzed.RESULTS In the period examined,44 stapes surgical procedures were performed and out of these 29 met the inclusion criteria.All patients were performed under otoendoscopy.There was no significant change in bone conduction threshold(P>0.05),but the average air conduction threshold improved from the preoperative(59.91±10.25)dB HL to(38.92±11.37)dB HL with air-bone gap reduced from the preoperative(37.37±7.65)dB HL to(18.71±8.38)dB HL(P<0.05).There were no serious complications such as sensorineural hearing loss and facial paralysis,but 22 patients were accompanied by varying degrees of vertigo,which were relieved after symptomatic treatment.CONCLUSION Endoscopic stapes surgery is a safe procedure with a low risk of peri-or postoperative complications,and can improve the air conduction hearing of otosclerosis patients.

Objective:To study the clinical characteristics of neonatal gastric perforation (NGP) and risk factors of mortality.Methods:From January, 2015 to December, 2021, clinical manifestations of neonates diagnosed with NGP in the Department of Neonatology and Neonatal Surgical Intensive Care Unit of our hospital were retrospectively analyzed. Neonates were assigned into the survival group and the death group according to their prognosis. Risk factors of mortality were analyzed using multivariate logistic regression method.Results:A total of 50 cases were enrolled, including 41 in the survival group and 9 in the death group. 38 cases were males, 34 were premature infants, 30 were low birth weight infants and 5 had history of asphyxia. The clinical manifestations included abdominal distension, tachypnea, cyanosis, poor response, fever, diminished bowel sound and redness of the abdominal wall. Abdominal X-ray indicated pneumoperitoneum. Laboratory abnormalities included leukocytosis, thrombocytopenia, elevated C-reactive protein and procalcitonin, decreased blood pH and increased lactic acid. 30 cases had perforation at the greater curvature of stomach. Perforation was larger than 3 cm in 40 cases and intestinal necrosis was identified in 14 cases. Some patients suffered from sepsis, respiratory failure, pulmonary hemorrhage, shock, coagulopathy and other related complications. The death group had significantly higher incidences of dyspnea, fever, elevated procalcitonin, blood pH<7.3, intestinal necrosis, time from onset of clinical manifestations to operation (Tm-o) >24 h and complications than the survival group ( P<0.05). Multivariate logistic regression analysis showed that pH<7.3 ( OR=9.755, 95% CI 1.363-69.800), Tm-o>24 h (OR=11.831, 95%CI 1.305-107.301), septic shock and sepsis ( OR=29.622, 95% CI 3.728-235.369) were risk factors of mortality. Conclusions:The main manifestations of NGP are abdominal distension and pneumoperitoneum. The risk factors of mortality in NGP are sepsis, blood pH<7.3 and Tm-o>24 h.

Objective:To study the risk factors and prognosis of pulmonary hypertension(PH) associated with bronchopulmonary dysplasia (BPD) in extremely preterm infants(EPIs).Methods:From January 2020 to December 2021, EPIs [gestational age (GA) <32 w] with BPD admitted to NICU of our hospital were retrospectively assigned into two groups: BPD with late-onset PH(PH group) and BPD without late-onset PH(non-PH group). Their general condition, treatment and prognosis were compared and the risk factors of late-onset PH were analyzed.Results:A total of 229 EPIs with BPD were enrolled, including 24(10.5%) in the PH group and 205(89.5%) in the non-PH group. The PH group had significantly smaller GA [(27.9±2.3) w vs. (28.7±1.8) w], longer mechanical ventilation [42.0(16.0, 84.0) d vs. 9.0(2.0, 23.0) d], longer hospital stay [100.5(86.3, 142.0) d vs. 77.0(56.5, 96.5)d],higher incidence of early-onset PH(54.2% vs. 9.3%) and higher mortality rate(33.3% vs. 9.8%) than the non-PH group ( P<0.05). Multivariate logistic regression analysis showed prolonged mechanical ventilation ( OR=1.046, 95% CI 1.011~1.064), early-onset PH ( OR=5.414, 95% CI 1.796~16.323) were independent risk factors for BPD with late-onset PH. 8(33.3%) patients in the PH group died, including 2 with grade Ⅱ BPD and 6 grade Ⅲ BPD. Conclusions:Prolonged mechanical ventilation and early-onset PH are independent risk factors for late-onset PH in BPD infants. BPD infants with late-onset PH have longer hospital stay, higher mortality and worse prognosis.