BACKGROUND:Chitosan is wel known as good biocompatibility and biodegradability;however, its extensive use in biomedical applications is restricted due to its poor transfection efficiency. OBJECTIVE:To prepare the polyethyleneimine-chitosan/DNA nanoparticles loading enhanced green fluorescent protein gene, and to detect their physicochemical properties and gene transfection efficiency towards chondrocytes in vitro.
METHODS:Low molecular weight polyethyleneimine was covalently linked to chitosan backbone to construct chitosan-graft-polyethyleneimine;then the chitosan-graft-polyethyleneimine was mixed with DNA nanoparticles, which loaded enhanced green fluorescent protein gene, by a complex coacervation method. The nanoparticle morphology was observed under a scanning electron microscopy. The sizes and zeta-potentials of the
nanoparticles were measured by a Marven-nano laser diffractometer. The binding capacity of plasmid DNA was evaluated by agarose gel electrophoresis analysis. The gene transfection experiments in vitro were performed towards rabbit’s chondrocytes. The gene transfection efficiency was measured with flow cytometry and under fluorescence microscope. How marked DNA entered into the nucleus of chondrocytes mediated by the nanoparticles was detected by laser scanning confocal microscopy.
RESULTS AND CONCLUSION:The prepared nanoparticles were mainly spherical, with an average size of (154.6±18.6) nm, and zeta-potential of (24.68±6.82) mV. The agarose gel electrophoresis analysis confirmed that the nanoparticles could effectively protect plasmid DNA from DNase Ⅰ-induced degradation. Gene transfection in vitro proved that the nanoparticles were efficient in transfecting rabbit’s chondrocytes and the expression of green fluorescent proteins was observed under fluorescent microscope, with a transfection efficiency of (23.80±1.74)%that was significantly higher than that of the naked plasmid DNA and chitosan/DNA nanoparticles (P<0.05). But no significant differences were observed between polyethyleneimine-chitosan/DNA nanoparticles and LipofectamineTM 2000. These findings indicate that the polyethyleneimine-chitosan/DNA nanoparticles can effectively protect plasmid DNA from nuclease degradation, and exhibit the favorable transfection ability towards articular chondrocytes.

Objective To establish the biological reference intervals of serum immunoglobulin ( Ig) for children ( neonates to young adults) in our laboratory.Methods This was a retrospective study.Serum IgG, IgA and IgM of the neonatal ( 1-28 days ) to 18 years old who visited the First Hospital of Jilin University during January 2011 to July 2014 were measured.The inclusion criteria were normal C-reactive protein content, normal liver and kidney function and without history of chronic diseases, allergic reactions, connective tissue diseases, rheumatic diseases and human immunodeficiency diseases.Children whose Ig tests were below upper limit of adult reference range were divided into 19 age groups.By eliminating outliers within each group, 9 466 cases of reference individuals conformed to the establishment of reference intervals were selected.Rank sum test was applied in comparing Ig levels of each two adjacent age groups, if there was not statistical significance ( P>0.05 ) , the two age groups were merged; otherwise not merged.The upper and lower limit of Ig reference range for each age group were calculated using nonparametric method, and biological reference intervals of children′s serum Ig were established.Results The neonatal serum IgG decreased grually after birth and reached the lowest point at 29 days-3 months, then the concentration increased gradually along with age growth and reached adult level by 11 years old.Neonatal serum IgA and IgM levels were at the lowest point and the concentration increased along with age growth then reached adult levels by 11 and 2 years old respectively.The children′s serum Ig reference intervals were as below:IgG:3.59-7.90, 2.26-5.40, 2.72-6.62, 2.87-7.74, 3.38-8.07, 3.80-9.08, 4.86-11.40, 4.97-11.70, 5.51-12.40, 6.17-13.10, 6.41-13.60, 6.53-14.20 and 6.84-14.30 g/L for neonatal, 29 days-3 months, 4-6 months, 7 months, 8-9 months, 10 months-1 year old, 2, 3, 4, 5-6, 7, 8 and 9-10 years old, respectively;IgA:0.23-0.45, 0.24-1.02, 0.23-0.79, 0.23-0.92, 0.24-1.03, 0.24-1.56, 0.26-1.93, 0.31-2.16, 0.44-2.56, 0.56-2.85, 0.52-3.35 and 0.63-3.23 g/L for neonatal-3 months, 4 months, 5-7 months, 8-9 months, 10 months-1 years old, 2, 3, 4, 5-6, 7, 8, and 9-10 years old, respectively;IgM:0.16-0.70, 0.21-1.20, 0.30-1.62, 0.38-2.16 and 0.44-2.17 g/L for neonatal, 29 days-3 months, 4-7 months, 8-9 months and 10 months-1 years old, respectively.Conclusions There were great differences of the serum Ig concentrations between children and adults, thus children′s immunologic function should not be assessed based on adult Ig reference intervals.According to the research, reference intervals of Ig were obtained for children at different age groups in the laboratory and reference in evaluating children′s immunologic function was provided for clinicians.

Objective: To compare the effectiveness and clinical safety between the two through the observation of Orthokeratology lens and multifocal soft lens. Methods: By using prospective longitudinal design, sixty patients (120 eyes) with orthokeratology lens and multifocal soft lens were selected. After 1, 3, 6 and 12 months, our tests were conducted, including ocular surface disease index (OSDI) questionnaire, tear break up time(BUT), corneal fluerescein staining (CSF) and meibomian gland function assessment, before and after wearing glasses, ocular axis and refraction in 6,12 and 18 months after were evaluated to assess the effect of myopia control. Results: But in the orthokeratology lens group was lower than the soft lens group after two months. The difference was statistically significant( t=4.35, P <0.05). After 1 month of wearing glasses, BUT in the orthokeratologylens group was shorter than before( F=305.83, P <0.05). In the group of soft lens, there was no statistically significance before and after wearing glasses( F= 0.38, P >0.05). There was no difference in corneal fluorescence staining (CSF) and meibomian gland function between the two groups before and after wearing glasses.( F time=1.55, 1.38, 10.15, 1.50, P >0.05, F group=2.31, 0.02， P >0.05). After 18 months of wearing orthokeratology lens, the average degree of spherical lens increased by -0.53 D, which was different from that before wearing( F=6.54, P <0.05). After wearing soft lens 18 month, the average degree of spherical lens increase by -0.34 D, which was different from that before wearing( F=2.88, P <0.05). After 18 months of wearing glasses, there were differences in the growth of ocular axis between the two groups( F =2.73, 2.83, P <0.05). Conclusion Orthokeratology lens and multifocal soft lens are both safe and effective measures to control myopia.

Objective:To observe and determine the gene mutation site and clinical phenotype of a NDP gene mutant family, and provide a basis for the prenatal diagnosis of offspring. Methods:A pedigree investigation study. Two patients and 6 family members of a third-generation Han family with NDP gene mutation who were admitted to the Maternal and Child Health Hospital of Gansu Province from July 2019 to December 2021 were included in the study. The patients and their parents underwent the examination of pupil light reflex, strip light imaging, visual acuity evaluation, fundus color photography, and wide-field fluorescein fundus angiography (FFA). Peripheral blood of all the subjects was collected, the pathogenic genes were screened by whole exome sequencing, and NDP genes were detected by amplification of multiple ligated probes. DNA prenatal diagnosis was performed by amniocentesis at 19th weeks of the mother's third gestation. Results:Proband (Ⅲ1), male, 4 years old, full term natural delivery. At about 40 days after birth, B-mode ultrasonography indicated total retinal detachment in both eyes. Normal hearing and intelligence. Fundus examination was not performed. First sibling of proband (Ⅲ2, big younger brother), ophthalmologic examination 30 days after birth, retinal detachment in both eyes. Proband's mother (Ⅱ2) had unvascularized peripheral temporal retina in both eyes. Wide-angle FFA examination showed no vascularization of the peripheral temporal retina in both eyes, and slight leakage of peripheral vascular fluorescein. The proband's second sibling (Ⅲ3, little younger brother) was screened for neonatal eye disease 1 day after birth. No abnormalities were observed outside both eyes. Cornea and lens transparent. No abnormalities were observed in the optic disc and macula in both eyes. No vascular curvature was observed in the peripheral retina. The results of gene detection showed that there was hemizygote deletion in exon 2 of NDP gene of the proband (Ⅲ1) and its big younger brother (Ⅲ2). His mother (Ⅱ2) had heterozygosity deletion in exon 2 of NDP gene. The phenotype and genetic test results of the proband's father (Ⅱ1), uncle (Ⅱ3), maternal grandfather (Ⅰ1) and maternal grandmother (Ⅰ2) were not abnormal. Conclusions:The hemizygote deletion in exon 2 of NDP gene is a pathogenic variation in the native family. The clinical phenotypes of different genders are different. Prenatal diagnosis is an effective way to block hereditary diseases in families.

Objective To explore the relationship between different smoking status and hypothyroidism in six iodine-suitable areas of China. Methods A total of 8187 residents were investigated by cluster sampling in six cities, and 7448 residents were included in the survey. The height, weight, waist circumference, and blood pressure were measured by filling out epidemiological questionnaire. Blood samples were collected to detect thyroid stimulating hormone ( TSH) , thyroid peroxidase antibody ( TPOAb) , and thyroglobulin antibody ( TgAb) . Results The mean TSH, TPOAb, and TgAb positive rates in passive smoking and active smoking groups were all lower than those in non-smoking group ( all P<0.01) . In the active smoking group, the TSH value decreased by 0.023 units for every unit increase in smoking index. The positive rates of TgAb and TPOAb in both passive smoking and active smoking groups were lower than those in non-smoking group (all P<0.01). Active and passive smoking reduced the prevalence of hypothyroidism (both P<0.01). Among women, the risks of clinical hypothyroidism and subclinical hypothyroidism were reduced in both active and passive smoking groups. Besides, the risk of subclinical hypothyroidism decreased significantly when the smoking index was more than 70. In male population, the risk of subclinical hypothyroidism in active and passive smoking group decreased. Besides, the risk of clinical hypothyroidism and subclinical hypothyroidism decreased significantly when the smoking index was more than 70. Conclusion Smoking in iodine-suitable areas may reduce TSH level and the positive rates of TPOAb and TgAb.

Objective:To explore the clinical and pathological characteristics and prognostic factors of gastric neuroendocrine carcinoma(G-NEC)and gastric mixed adenoendocrine carcinoma(G-MANEC).Methods:Retrospective analysis was conducted on the clinical data of 67 patients with G-NEC and G-MANEC who underwent surgical treatment at Heping Hospital Affiliated to Changzhi Medical College from May 2015 to May 2023.The study included an analysis of the pathological characteristics distinguishing G-NEC from G-MANEC.Results:Com-pared to gastric adenocarcinoma,patients with G-NEC and G-MANEC in the stomach showed a higher incidence of gastric cancer in the male gastric cardia and were diagnosed at a later age.Tumors with larger diameters increase susceptibility to anemia,low albumin levels,and in-vasion of nerves and vasculature.Deeper tumor infiltration is associated with increased local lymph node metastases,later TNM staging,and a higher likelihood of distant metastasis post-surgery.The prognosis of G-NEC and G-MANEC is worse than that of gastric adenocarcinoma(P=0.001).However,there is no statistically significant difference in the pathological characteristics(P>0.05)and prognosis analysis(P=0.212)between G-NEC and G-MANEC.Univariate survival analysis identified age,preoperative albumin,preoperative CEA,number of lymph node metastases,TNM staging,and postoperative distant metastasis as risk factors affecting patient's overall survival(OS).In the multivariate ana-lysis,age,preoperative albumin,TNM staging,and postoperative distant metastasiswere identified as independent risk factors for OS.Con-clusions:There is a significant difference in clinical characteristics between G-NEC,G-MANEC,and gastric adenocarcinoma,often diagnosed at an advanced stage,which is prone to distant metastasis post-surgery.Poor prognosis is observed in patients aged over 60 years,with pre-operative albumin<40g/L,TNM stage Ⅱ/Ⅲ,and postoperative distant metastasis.