Objective To explore the application value of anhydrous ethanol as an alternative to methanol in the preparation of chromosomal specimens from peripheral blood lymphocytes, and to establish a set of quantitative analytical methods for objectively evaluating the effectiveness of specimen preparation. Methods Residual blood samples from routine laboratory slide preparation were used for lymphocyte culture. The standard slide preparation method was employed. The fixative in the control group was methanol and glacial acetic acid (3∶1). Four experimental groups were set up based on the ratio of anhydrous ethanol to glacial acetic acid in the fixative (volume ratios of 3∶1, 5∶1, 7∶1, and 9∶1 for experimental groups 1, 2, 3, and 4, respectively). A chromosomal analysis was conducted using an automated chromosome scanning/image analysis system to evaluate the morphology and dispersion of metaphase chromosomes in both control and experimental groups. Comparisons were made between the control and experimental groups regarding the dic + r aberration rate, ace aberration rate, chromosomal aberration rate, chromosome dispersion index, chromosome overlapping ratio, and dispersion index/overlapping ratio. Results Microscopic evaluation revealed that the preparation quality of experimental groups 1 and 2 was comparable to the control group. No statistically significant differences were observed in dic + r aberration rate between each of the experimental groups and the control (P > 0.05). All experimental groups except group 4 showed no significant differences in ace aberration rate and chromosome aberration rate compared with the control group (P > 0.05). Experimental groups 1 and 2 showed no significant differences in chromosome dispersion index, overlapping ratio, and dispersion index/overlapping ratio compared with the control group (P > 0.05). Conclusion A mixture of anhydrous ethanol and glacial acetic acid at a 5∶1 ratio is recommended for use as a fixative in the preparation of chromosomal specimens from peripheral blood lymphocytes. A quantitative index system for assessing the quality of chromosomal specimens was established, enabling objective evaluation of slide preparation effectiveness.

Objective To analyze the relationship between abnormal expression of serum neurohemoglobin(NGB)and myelin basic protein(MBP)and the cerebral neurodevelopment of neonates with hypoxic-ischemic encephalopathy(HIE).Methods A total of 89 preterm infants with HIE admitted in the hospital between January 2023 and March 2024 were selected as the observation group,and 60 preterm infants without HIE during the same period were selected as the control group.Serum levels of MBP,NGB and secretagogues in two groups were detected,and the neonates amplitude integration electroencephalogram score was evaluated.The neurological function of neonates was evaluated using 20 items of Neonatal Behavioral Neurological As-sessment(NBNA).The correlation test and diagnostic value were evaluated using Spearman method and re-ceiver operating characteristic(ROC)curve.Results The serum levels of NGB,MBP and secretagogue in the ob-servation group were higher than those in the control group(P<0.05),and the amplitude integration electroencepha-logram score was lower than that in the control group(P<0.05).The serum MBP,NGB and secretagogue levels in the mild,moderate,and severe groups increased sequentially(P<0.05),NBNA score and amplitude integration e-lectroencephalogram score decreased sequentially(P<0.05).The levels of NGB,MBP and secretagogue were the risk factors affecting NBNA score(P<0.05),and the amplitude integration electroencephalogram score was a protective factor affecting NBNA score(P<0.05).The area under the curve of NGB and MBP in diag-nosing HIE was greater than 0.8,which had high application value.Conclusion Serum NGB and MBP levels are closely related to the severity of HIE,and have certain connection with NBNA score.Elevated levels of NGB and MBP in neonates with HIE may be related to the body's stress response to nerve damage,which could reflect to some extent the brain nerve function damage in with HIE.

Immunotherapy has become the first-line standard treatment option for driver gene-negative advanced non-small cell lung cancer (NSCLC). But not all patients can benefit from immunotherapy, and can even have serious adverse reactions. It is crucial to identify the predictors of clinical response to immunotherapy. Several studies have shown that elevated baseline C-reactive protein (CRP) or persistent elevation of CRP during the treatment process may indicate a poorer prognosis for patients, and high CRP may be correlated with adverse reactions. Attention to the dynamic changes of CRP during immunotherapy for NSCLC may become an important predictor of prognosis.

Objective:To investigate the changes in gray matter volume (GMV) and abnormalities in structural covariant network (SCN) patterns in patients with chronic pontine infarction (PI).Methods:Patients with unilateral chronic PI (case group) with the first onset admitted to the First Affiliated Hospital of Zhengzhou University and Tianjin Medical University General Hospital from October 2014 to June 2021 were prospectively included. At the same time, healthy subjects matched with age, gender and education years (normal control group) were included. High-resolution three-dimensional T 1 structural MRI images and behavioral scores of the subjects were collected. The voxel-based morphometry and two-sample t test were used to explore the differences in GMV between the groups. Using GMV differential brain regions as seed points, SCN was constructed to explore the abnormality of structural covariant patterns in patients with PI. Spearman rank correlation analysis was used to analyze the correlation between GMV in differential brain regions and behavioral scores. Results:A total of 60 patients with PI were enrolled, including 33 left PI and 27 right PI, while 34 healthy controls were also enrolled. Compared with the normal control group, the GMV in bilateral posterior cerebellar lobe decreased significantly in the left PI group, and the GMV in left anterior and posterior cerebellar lobes and the right posterior cerebellar lobe decreased significantly in the right PI group (Gaussian random field correction with voxel level P<0.001 and cluster level P<0.05, cluster voxel >20), and there was a significant correlation between GMV values in the left anterior and posterior cerebellar lobes and the right posterior cerebellar lobe and the motor function score ( P<0.05). In addition, compared with the normal control group, the right PI group had broader covariate brain regions and a significant increase in the number of structural connections between covariate brain regions (family-wise error correction with voxel level P<0.05, cluster voxel >20). Conclusions:The GMV in bilateral posterior cerebellar lobe decreases significantly in patients with chronic PI, and were secondary to broader covariate brain regions and structural connections. This may be the neural mechanism of impaired behavioral function in patients with PI.

Objective:To evaluate the applicability of the Zhang′s diagnostic criteria for atopic dermatitis (AD) in adolescents and adults from southern Jiangsu.Methods:A total of 1 769 patients were collected, who were newly diagnosed with eczema or AD in departments of dermatology of 7 hospitals in southern Jiangsu from May 2019 to May 2021. A standardized survey interview was conducted with patients by dermatologists using a self-designed questionnaire, which included patient′s personal information, relevant medical history, clinical characteristics, laboratory tests, etc. Using Hanifin-Rajka criteria as the gold standard, the sensitivity and specificity of the Williams criteria, Zhang′s criteria and Japanese Dermatological Association (JDA) criteria were evaluated, separately.Results:Among the 1 769 patients, there were 759 (42.9%) males and 1 010 (57.1%) females, aged 32.2 ± 8.2 years (range, 12 - 79 years) . Pruritus was the most common clinical feature among the AD patients (883/913, 96.7%) . When using the Hanifin-Rajka criteria as the gold standard, 913 (51.6%) subjects were diagnosed with AD; the sensitivity of Zhang′s criteria reached 92.6% (845/913) , and its specificity was 73.2% (627/856) ; the sensitivity of Williams′ criteria was 87.8% (802/913) , and its specificity was 81.3% (696/856) ; the sensitivity of JDA criteria reached 96.9% (885/913) , and its specificity was 68.9% (590/856) . The consistency in the diagnosis of AD was moderate between the Zhang′s criteria and Williams criteria (Kappa = 0.61, P = 0.009) , was relatively high between the Zhang′s criteria and JDA criteria (Kappa = 0.85, P = 0.001) , and was moderate between the Williams criteria and JDA criteria (Kappa = 0.51, P = 0.013) . Conclusions:Compared with the Hanifin-Rajka criteria, the Zhang′s criteria exhibit good sensitivity and specificity for the diagnosis of AD among adolescents and adults in southern Jiangsu. However, pruritus remains important for the diagnosis of AD.

With the development of detection methods, various biomarkers of liver cancer have been detected constantly, which is of great significance for the early diagnosis and real-time monitoring of liver cancer after treatment. Based on the differences in the sensitivity and specificity of different biomarkers, exploration of the value of diverse biomarkers in the diagnosis and prognosis assessment of liver cancer can provide an important reference for clinicians to scientific and rational application of distinct biomarkers.

Objective:To evaluate the efficacy and safety of recombinant human tumor necrosis factor-α receptorⅡ: IgG Fc fusion protein (rhTNFR:Fc) in the treatment of drug-induced toxic epidermal necrolysis (TEN) .Methods:From 2009 to 2018, 22 patients with TEN were enrolled from 8 centers such as the Second Affiliated Hospital of Soochow University, including 10 males and 12 females, whose age ranged from 22 to 75 years. These patients were subcutaneously injected with rhTNFR:Fc at a dose of 25 mg once every 3 days for 6 - 8 consecutive sessions, and the initial dose was doubled. The drug eruption area and severity index (DASI) score and DASI improvement indices (DASI50, DASI75 and DASI90) were assessed before treatment and on days 4, 7, 10, 13, 16, 19, 22 and 25 after treatment; cytometric bead array (CBA) technology was used to detect the level of tumor necrosis factor (TNF) -α in peripheral blood and blister fluid samples. During the treatment, body temperature, rash changes, liver and kidney function of patients were monitored, and adverse reactions were recorded. Statistical analysis was carried out by using repeated measures analysis of variance, paired t test and Pearson correlation analysis. Results:Of the 22 patients, the temperature stopped rising in 20 patients without infections 24 - 72 hours after the first treatment, and returned to normal after 48 - 120 hours. Among the 22 patients, new blisters stopped appearing 24 - 48 hours after the first treatment, the skin color changed from bright red to dark purple after 48 - 96 hours, and most skin lesions subsided after 2 weeks. After 2 - 4 weeks of treatment, levels of alanine aminotransferase and aspartate aminotransferase returned to normal in 19 patients with abnormal liver function. After 4 - 13 days of treatment, levels of creatinine and urea nitrogen stopped rising in 7 patients with abnormal renal function. During the treatment, the DASI score of the 22 patients gradually decreased ( F = 532.81, P < 0.01) , from 53.64 ± 8.67 before treatment to 2.05 ± 1.21 on day 25 after treatment ( t = 26.60, P < 0.001) . On day 10 after treatment, 22 patients (100%) achieved DASI50; on day 19, 22 (100%) achieved DASI75; on day 25, 20 (90.90%) achieved DASI90. The level of TNF-α in peripheral blood of the 22 patients gradually decreased along with the extension of treatment duration, from 33.95 ± 27.90 ng/L before treatment to 2.38 ± 0.79 ng/L on day 25. Before treatment, the level of TNF-α in blister fluid of 15 patients was 111.99 ± 99.41 ng/L, and the ratio of blister-fluid TNF-α level to peripheral blood TNF-α level was 1.83 - 28.21. Before treatment, no correlation was observed between the serum level of TNF-α and DASI score in the 22 patients ( P = 0.10) , while the blister-fluid TNF-α level was positively correlated with DASI score in the 15 patients ( r = 0.59, P = 0.02) . No acute adverse reactions were observed during the treatment. All the 22 patients completed the treatment and were discharged with complete recovery. During 6 months of follow-up after discharge, no recurrence or any complication was observed. Conclusion:rhTNFR:Fc is effective and safe for the treatment of drug-induced TEN.

Objective: To discuss the impact and practicability of pluralistic management when puerpera with hepatitis B virus (HBV) in hospital. Methods: By review researching and convenient sampling, two groups of puerpera with HBV were studied comparatively by the index of the time of breast milk, the breastfeeding rate and the satisfaction rate in hospital.The control group (411 cases, January to December, 2017) carried on the normal regulations nursing and the observation group (411 cases, January to October, 2018) carried on the pluralistic management in the Obstetrics and Gynecology Department of the Fifth Medical Center of PLA General Hospital. Results: The rate of the starting time of milk secretion (<24, 24-48, 49-72, >72 h) was 29.4%(121/411), 36.0%(148/411), 24.6%(101/411), 1.0%(41/411) in the observation group, and 13.6%(56/411), 21.7%(89/411), 33.3%(137/411), 31.4%(129/411) in the control group. The differences between the two groups were significant (χ² value was 7.66-57.43, all P<0.01). Postpartum hospital stay in 3 days and 6 days, the breastfeeding rate and average rate was 90.4%(198/219), 95.3%(183/192), 92.7%(381/411) in the observation group, and 79.0%(180/228), 84.2%(154/183), 81.3%(334/411) in the control group, the differences between the two groups were significant (χ² value was 11.13, 12.7, 23.61, all P<0.01). The total satisfaction rate in hospital was 99.5%(409/411) in the observation group, and 91.2%(375/411) in the control group, the difference between the two groups was significant (χ² value was 31.93, P<0.01). Conclusions Pluralistic management can raise the rate of breastfeeding of puerpera with HBV and the satisfaction of puerpera with HBV and her family.

Objective To study the expression and significance of Nrf2 and caspase-3 in non-small cell lung cancer (NSCLC).Methods The specimens from 82 cases of NSCLC in our hospital from September 2015 to December 2017 were selected for observation.All patients were surgically resected to obtain NSCLC specimens.The expression of Nrf2 and caspase-3 in NSCLC was detected by immunohistochemistry and its clinical significance was analyzed.Results The positive expression rate of Nrf2 in NSCLC (63.4％) was significantly higher than that in normal tissue (30.0％),with statistically significant difference (P ＜ 0.05).The analysis of clinical characteristics showed that the expression of Nrf2 in NSCLC was related to tumor diameter,tumor node metastasis (TNM) stage,lymph node metastasis and distant metastasis (P ＜ 0.05).The expression of Nrf2 in NSCLC was not significantly associated with gender,age,smoking history,case classification,and histological differentiation (P ＞ 0.05).The positive rate of caspase-3 in NSCLC (40.2％) and the positive rate of caspase-3 in normal tissues (40.0％) were not statistically different (P ＞ 0.05).Analysis of clinical characteristics showed that the expression of caspase-3 in NSCLC was related to TNM stage,lymph node metastasis,distant metastasis,and the difference was statistically significant (P ＜ 0.05);the expression of Nrf2 in NSCLC was not significantly associated with gender,age,smoking history,case classification,tissue differentiation,and tumor diameter (P ＞ 0.05).Conclusions The abnormal expression of Nrf2 and caspase-3 is correlated with the progression of NSCLC and the malignant biological behavior.Active detection of Nrf2 and caspase-3 levels can clinically determine the clinical stage,lymph node metastasis and distant metastasis of NSCLC,providing information to facilitate clinical development of relevant interventions.

Objective: To elucidate the mechanism of melatonin combined with cisplatin in promoting cell apoptosis of rat pancreatic cancer AR42J cells. Methods: Rat pancreatic cancer AR42J cells were divided into control group, 1 mmol/L cisplatin treated group (cisplatin group), 1 mmol/L melatonin treated group (melatonin group), 1 mmol/L cisplatin combined with 1 mmol/L melatonin treated group (combined group), 1 μmol/L cisplatin combined with melatonin treated group after 1 μmol/L PBN pretreatment for an hour (PBN+ combined group) and 1 μmol/L cisplatin combined with melatonin treated group after PBN solvent pretreatment for an hour (solvent+ combined group). MTT and annexin V-FITC/PI were used to detect the cell proliferation rate and cell apoptosis rate, respectively. The protein expression of caspase-3 was detected by Western blot. DCFH-DA was used to detect the level of ROS. ROS level and caspase-3 expression in AR42J cells pretreated with ROS antagonist PBN for 24 hours were detected. Results: The cell proliferation rate of control group, cisplatin group, melatonin group and combination group after 24-hour culture was (96.29±3.49)%, (81.38±6.01)%, (80.72±3.68)% and (42.26±6.35)%, respectively. The cell apoptosis rate was (16.42±4.15)%, (56.47±9.06)%, (52.94±6.57)% and (87.36±6.48)%, respectively. The percentage of ROS positive cells was (1.33±1.53)%, (46.67±7.64)%, (45.67±5.13)% and (83.33±7.64)%, respectively. The expression of cspase-3 was 100%, (150.64±7.70)%, (147.00±7.27)% and (190.04±5.07)%, respectively. The cell proliferation rate of cisplatin group and melatonin group was significantly lower than that of the control group. The apoptotic rate, the proportion of ROS positive cells and the expression of caspase-3 were significantly higher than those in control group. The changes in the combined group were more obvious than those in the single drug treatment group, and the differences were all statistically significant (all Pvalues <0.05). The percentage of ROS positive AR42J cells [(45.24±4.64)% vs (84.90±6.13)%)] induced by melatonin combined with cisplatin could be significantly reversed by PBN pretreatment for 1 hour, and the expression of caspase-3 in AR42J cells could be down regulated (124.92±9.72 vs 184.69±7.76) with statistically significant difference (all P values<0.05). Conclusions Melatonin combined with cisplatin may promote cell apoptosis of pancreatic cancer AR42J cells through ROS/caspase-3 pathway.