Objective To investigate the effects of miR-135a on HOXA10 expression,proliferation and apoptosis of SKOV3 cells.Methods (1) Through computer-aided algorithms,the predicted target gene of miR-135a (HOXA10)were determined.(2) miR-135a mimics,miR-135a inhibitor and negative control were transfected into SKOV3 cells,respectively.Reverse transcription (RT)-PCR,western blot analysis were used to examine the expression levels of HOXA10 at different times (24,48 and 72 hours).(3) A luciferase reporter assay was used to confirm the direct regulation between miR-135a and HOXA10.(4) SKOV3 cells proliferation at different times (24,48 and 72 hours) was detected by methyl thiazolyl tetrazolium(MTT) assay [quantified by absorbance(A)].Western blot was used to examine the expression of apoptosis-associated protein bcl-2,bax and caspase-3 in SKOV3 cells after 48 hours transfection.Results (1) HOXA10 was predicted to be the target gene of miR-135a by computer-aided algorithms.(2) RT-PCR shown that HOXA10 mRNA levels were decreased over time (24,48 and 72 hours) after miR-135a mimics transfectionin SKOV3 cells (0.94 ±0.04 vs 0.78 ±0.03 vs 0.70 ±0.03,P ＜0.05).While,the expression of HOXA10 mRNA was increased over time after miR-135a inhibitor transfection (1.14 ± 0.05 vs 1.16 ±0.03 vs 2.60 ±0.08,P ＜0.05).After transfected with miR-135a mimics or miR-135a inhibitor over 48 and 72 hours,the HOXA10 expression levels in SKOV3 cells were significantly lower or higher than each control group,respectively (all P ＜ 0.01).Western blot analysis of HOXA10 expression in SKOV3 cells confirmed the results of RT-PCR detected.(3) After cotransfection of miR-135a plasmid and pMIR-REPORT luciferase plasmid containing HOXA10,luciferase reporter assays showed that the luciferase activity reduced by 67.8％ (P ＜0.01).(4) MTT showed that SKOV3 cells growth after miR-135a mimics transfection for 48 and 72 hours were significantly lower than those in control group (0.38 ± 0.03 vs 0.52 ± 0.05,0.67 ±0.05 vs 0.75 ± 0.06 ; respectively,all P ＜ 0.05).While,SKOV3 cells transfected with miR-135a inhibitor for 72 hours grew significantly faster than that in control group (0.95 ± 0.05 vs 0.75 ± 0.06,P ＜ 0.01).After miR-135a mimics transfection,the level of bcl-2 protein was significantly lower than that in control group (0.28 ±0.06 vs 0.76 ±0.09,P ＜0.01).The activity of caspase-3 was significantly higher than that in control group (115.0 ± 2.4 vs 95.4 ± 2.1,P ＜ 0.01).While,there was no statistical difference of bax expression (P =0.142).However,after miR-135a inhibitor transfection,the expression level of bcl-2 protein was significantly higher than that in control group (0.92 ± 0.03 vs 0.76 ± 0.09,P =0.037) and the activity of caspase-3 was significantly lower than that in control group (59.5 ± 4.1 vs 95.4 ± 2.1,P ＜ 0.01).There was also no statistical difference of bax expression (P =0.066).Conclusion miR-135a may play an important role in cell proliferation and apoptosis of ovarian cancer cells by regulating HOXA10 and its downstream pathways.

Objective Amyotrophic lateral sclerosis(ALS)is a progressive paralytic disorder resulting from the degeneration of upper and lower motor neurons.Sporadic ALS(SALS)accounm for majority of patients.ALS is a kind of complex disorder.There were several single nucleotide polymorphism (SNP)reported to be associated with SALS in recently published genome-wide association(GWA)study,but there are few data from Asia ALS population and no report focus on SNP which may associated with SALS of Chinese origin.Our study is to screen and add the SNPs related to the risks of SALs in Chinese.Methods Eighty-six individuals with SALS and 94 matched controls were recruited for our study and genomic DNA from blood samples was extracted.Genotypes were determined by a matrix assisted laser desorption/ionization time of flisht mass spectrometry based approach followed by association analysis. Results Individual genotype data for 8 SNPs,rs6700125,rs10260404, rs1942239,rs2279812,rs2405657,rs558889,rs6922711 and rs935 1470 in Chinese population showed no significant association with sporadic ALS.Combining genotype data from published GWA,rs1942239 gained in strength of allelic association(P value decreased to 9.07×10-5 from 1.48×10-4),and rs558889 deviated Hardy-Weinberg equilibrium at ALS case group which may be associated with susceptibility.Conclusions SNP rs1942239 and rs558889 may contribute to susceptibility of sporadic ALS in Chinese patient.The larger sample studies are warranted to confirm the association.

Objective:To compare the detection and amplitude of epileptiform discharges (EDs) between surface sphenoidal electrode and anterior temporal electrode in patients with interictal EDs in the temporal region, and to explore the value of surface sphenoidal electrode.Methods:A total of 1 356 outpatients with epilepsy who underwent 2-hour video electroencephalogram (EEG) monitoring in Xuanwu Hospital from October to December 2021 were retrospectively enrolled. All patients were hooked up with scalp electrode according to the international 10-20 system as well as surface sphenoidal electrode and anterior temporal electrode. The EEGs with EDs recorded by surface sphenoidal electrode and/or anterior temporal electrode were selected for analysis. The detection rate and the amplitude of EDs by surface sphenoidal electrode and anterior temporal electrode were compared.Results:Seventy-three EEGs were collected and 250 EDs were counted. The detection rate of the anterior temporal electrode and surface sphenoidal electrode were 88.0% (220/250) and 98.4% (246/250) respectively. The difference in detection rate was statistically significant (χ 2=18.38, P<0.001). For the EDs from anterior temporal regions (taking the discharges recorded by anterior temporal electrode as "gold standard"), the detection rate of surface sphenoidal electrode was 98.2% (216/220). There was no statistically significant difference in detection rate between the anterior temporal electrode and surface sphenoidal electrode (χ 2=2.27, P=0.132). There were 216 EDs recorded by these two kinds of electrode simultaneously. The average amplitude of the EDs on surface sphenoidal electrode and anterior temporal electrode was (77.1±38.9) μV and (80.2±44.9) μV, respectively. The difference was statistically significant ( t=2.28, P=0.031). Conclusions:The detection rate of surface sphenoid electrodes was higher than that of anterior temporal electrodes for the EDs in the temporal region, and surface sphenoidal electrodes can be used routinely in outpatient. The surface sphenoidal electrode had more chance to detect EDs originating from regions out of the anterior temporal regions.

Objective:To explore the establishment and application of ovarian cancer organoids.Methods:Fresh ovarian tumor tissues, obtaining from patients underwent surgery in the First Affiliated Hospital of Nanjing Medical University between October 2021 and March 2022, were collected, enzymatic degraded, digested, and embedded into matrigel to establish organoids. A total of 32 ovarian cancer samples were collected. Hematoxylin eosin (HE) staining and immunofluorescence (IF) procedure were used to verify the morphological structure of organoids and their expression of molecular markers. 3D cyto-live or dead assay was used to detecte the live or dead cells in organoids. Carboplatin with a concentration ranging from 5 to 80 μmol/L (5, 10, 20, 40, 80 μmol/L) was added to organoids to calculate the 50% inhibitory concentration (IC 50) in different organoids. Results:(1) Organoids from a total of 32 patients were established, of which 18 cases could be passaged stably in the long term in vitro, while 14 could be passaged in the short time. The average amplification time of long-term passage in vitro was over 3 months, and the longest reached 9 months. (2) In HE staining, significant nuclei atypia and local micropapillary structures were observed in organoids. IF staining revealed that ovarian cancer organoids expressed molecular markers similar to primary tumor tissues, such as Pan cytokeratin (Pan-CK), p53, paired box gene 8 (PAX8), and Wilms tumor gene 1 (WT1). (3) In 3D cyto-live or dead assay, a large number of apoptotic cells were observed inside and around the organoids after added carboplatin. The sensitivity to carboplatin varied in 18 organoids could amplify in the long term, with an average IC 50 of (29.5±15.8) μmol/L. Moreover, IC 50 values of 4 organoids derived from patients received neoadjuvant chemotherapy were much higher than the 14 organoids which did not received neoadjuvant chemotherapy [(48.7±11.3) μmol/L vs (24.0±12.1) μmol/L; t=3.429, P=0.022]. Conclusions:Organoids recapitulate ovarian cancers in vitro and could be stably passaged. Organoids derived from patients received neoadjuvant chemotherapy have higher resistance to carboplatin.

Brain ; physiology ; Deep Brain Stimulation ; methods ; Epilepsy ; therapy ; Humans

Objective Spike and wave index (SWI) is of great importance for the diagnosis of electrical status epilepticus during sleep (ESES) in children patients. This study intended to compare four commonly used SWI methods with full night SWI to evaluate their performances. Methods Fifteen patients diagnosed with ESES by electroencephalogram initial report (SWI≥50%) were retrospectively analyzed,and SWI was calculated for the full night sleep(SWI),the first sleep cycle (SWI1),the first 5 minutes of the first sleep cycle (SWI15 min),the first 10 minutes (SWI10 min) and the first 15 minutes (SWI15 min). Wilcox paired test was used to compare SWI with other four SWI methods respectively. Results The SWI of all four methods was significantly different from the full night SWI (p-values of 0.003、0.002、0.002和0.001,respectively,α=0.0125). Among the 15 patients,14 (93.3%) had the trend that SWI decreased with sleep cycles overnight. Conclusion None of the four methods of SWI is a good substitute for full night SWI. The SWI tends to decrease with sleep cycles overnight,and using the first sleep cycle or anepoch of it to calculate SWI is likely to be overestimated.

Background: Thyroid diseases are highly prevalent worldwide, but their diagnosis remains a challenge. We established reference intervals (RIs) for thyroid-associated hormones and evaluated the prevalence of thyroid diseases in China. Methods: After excluding outliers based on the results of ultrasound screening, thyroid antibody tests, and the Tukey method, the medical records of 20,303 euthyroid adults, who visited the Department of Health Care at Peking Union Medical College Hospital from January 2014 to December 2018, were analyzed. Thyroid-associated hormones were measured by the Siemens Advia Centaur XP analyzer. The RIs for thyroid-associated hormones were calculated according to the CLSI C28-A3 guidelines, and were compared with the RIs provided by Siemens. The prevalence of thyroid diseases over the five years was evaluated and compared using the chi-square test. Results: The RIs for thyroid stimulating hormone (TSH), free thyroxine (FT4), free triiodothyronine (FT3), total thyroxine (TT4), and total triiodothyronine (TT3) were 0.71–4.92 mIU/L, 12.2–20.1 pmol/L, 3.9–6.0 pmol/L, 65.6–135.1 nmol/L, and 1.2–2.2 nmol/L, respectively. The RIs of all hormones except TT4 differed significantly between males and females. The RIs of TSH increased with increasing age. The prevalence of overt hypothyroidism, overt hyperthyroidism, subclinical hypothyroidism, and subclinical hyperthyroidism was 0.5% and 0.8%, 0.2% and 0.6%, 3.8% and 6.1%, and 3.3% and 4.7% in males and females, respectively, which differed from those provided by Siemens. Conclusions Sex-specific RIs were established for thyroid-associated hormones, and the prevalence of thyroid diseases was determined in the Chinese population.

Objective:To analyze the molecular epidemiological characteristics of the Corona virus disease 2019 (COVID-19) cases in Shijiazhuang, which can reveal the origin of the outbreak and provide a scientific basis for COVID-19 prevention and control.Methods:From January 2 to January 8, 2021, a total of 404 samples from 170 COVID-19 cases were collected from the Shijiazhuang Fifth Hospital. The consensus sequence of 2019 novel Coronavirus(2019-nCoV) was obtained through multiplex polymerase chain reaction-based sequencing. The sequences of 170 COVID-19 cases were analyzed by the PANGOLIN, and the data were statistically analyzed by T-test.Results:Among the 404 COVID-19 samples, a total of 356 samples obtained high quality genome sequences (>95%,100×sequencing depth). The whole genome sequences of 170 COVID-19 cases were obtained by eliminating repeated samples. All 170 sequences were recognized as lineage B1.1 using PANGOLIN. The number of single nucleotide polymorphism arrange from 18-22 and most of the single nucleotide polymorphism were synonymous variants. All of 170 genomes could be classified into 48 sub-groups and most of the genomes were classified into 2 sub-groups (66 and 31, respectively).Conclusions:All cases in this study are likely originated from one imported case. The viruses have spread in the community for a long time and have mutated during the community transmission.

To summarize the nursing experience of an adolescent with early repolarization syndrome who had implanted a subcutaneous implantable cardioverter defibrillator(ICD)and experiences frequent ventricular fibrillation.The main nursing care were as follows:recognizing a ventricular fibrillation attack and emergency care;risk prevention and handling in transportation;prevention of iatrogenic infection and strengthening protective isolation;promoting the recovery of brain function;personalized health education;pre-hospital first aid skills training for families.After careful treatment and nursing,the patient did not recur arrhythmia,and heart function had improved significantly,and the pulmonary inflammation disappeared.No medical-acquired infections occurred during hospitalization.The patient's motor ability and language expression ability recovered to the baseline,and was discharged on the 20th day of admission.A month after discharge,the patient recovered well and could study and live normally.

Objective:To study the distribution and antibiotics resistance of the main pathogens of neonatal purulent meningitis in different regions of China.Methods:A retrospective descriptive clinical epidemiological study was conducted in children with neonatal purulent meningitis which admitted to 18 tertiary hospitals in different regions of China between January 2015 to December 2019. The test results of blood and cerebrospinal fluid, and drug sensitivity test results of the main pathogens were collected. The distributions of pathogenic bacteria in children with neonatal purulent meningitis in preterm and term infants, early and late onset infants, in Zhejiang Province and other regions outside Zhejiang Province, and in Wenzhou region and other regions of Zhejiang Province were analyzed. The chi-square test was used for statistical analysis.Results:A total of 210 neonatal purulent meningitis cases were collected. The common pathogens were Escherichia coli ( E. coli)(41.4%(87/210)) and Streptococcus agalactiae ( S. agalactiae)(27.1%(57/210)). The proportion of Gram-negative bacteria in preterm infants (77.6%(45/58)) with neonatal purulent meningitis was higher than that in term infants (47.4%(72/152)), and the difference was statistically significant ( χ2=15.54, P=0.001). There were no significant differences in the constituent ratios of E. coli (36.5%(31/85) vs 44.8%(56/125)) and S. agalactiae (24.7%(21/85) vs 28.8%(36/125)) between early onset and late onset cases (both P>0.05). The most common pathogen was E. coli in different regions, with 46.7%(64/137) in Zhejiang Province and 31.5%(23/73) in other regions outside Zhejiang Province. In Zhejiang Province, S. agalactiae was detected in 49 out of 137 cases (35.8%), which was significantly higher than other regions outside Zhejiang Province (11.0%(8/73)). The proportions of Klebsiella pneumoniae, and coagulase-negative Staphylococcus in other regions outside Zhejiang Province (17.8%(13/73) and 16.4%(12/73)) were both higher than those in Zhejiang Province (2.9%(4/137) and 5.1%(7/137)). The differences were all statistically significant ( χ2=14.82, 12.26 and 7.43, respectively, all P<0.05). The proportion of Gram-positive bacteria in Wenzhou City (60.8%(31/51)) was higher than that in other regions in Zhejiang Province (38.4%(33/86)), and the difference was statistically significant ( χ2=6.46, P=0.011). E. coli was sensitive to meropenem (0/45), and 74.4%(32/43) of them were resistant to ampicillin. E. coli had different degrees of resistance to other common cephalosporins, among which, cefotaxime had the highest resistance rate of 41.8%(23/55), followed by ceftriaxone (32.4%(23/71)). S. agalactiae was sensitive to penicillin, vancomycin and linezolid. Conclusions:The composition ratios of pathogenic bacteria of neonatal purulent meningitis are different in different regions of China. The most common pathogen is E. coli, which is sensitive to meropenem, while it has different degrees of resistance to other common cephalosporins, especially to cefotaxime.