The major bottle-neck in the way of constructing high density protein microarray is the availability and stability of proteins. The traditional methods of generating protein arrays require the in-vivo expression, purification and immobilization of hundreds or thousands of proteins. The cell-free protein array technology employs cell-free expression systems to produce proteins directly onto surface from co-distributed or pre-arrayed DNA or RNA, thus avoiding the laborious and often costly processes of protein preparation in the traditional approach. Here we provide an overview of recently developed novel technology in cell free based protein microarray and their applications in protein interaction analysis, in antibody specificity and vaccine screening, and in biomarker assay.
Cell-Free System ; DNA ; genetics ; Humans ; Protein Array Analysis ; methods ; Protein Biosynthesis ; Protein Interaction Mapping ; Proteins ; analysis ; genetics ; metabolism

Objective To investigate the effect of comprehensive nursing intervention and evaluate its role in the entire clinical course of in the treatment of children with severe burns. Methods A total of 60 children with 1-5 years old with clinical diagnosis of severe burns were randomly divided into experimental group and control group by random digits table method, 30 cases in each group. The control group received routine burn care. The experimental group received comprehensive nursing interventions on the basis of routine burn care and the intervention time covered the entire course of treatment. During hospitalization, the recovery and family social system were evaluated by anxiety scales and social support scales combined with the cure rate and hospitalization of children. Results Before intervention, the score of Anxiety Rating Scale was high in both groups of families, while Social Support Scale was lower, the difference was not statistically significant between the two groups (P>0.05). After intervention, the score of Anxiety Rating Scale and was Social Support Scale were (52.31±7.81), (31.52± 5.48)points in the experimental group, and (62.51 ± 6.52), (23.62 ± 5.62) points in the control group, and there were significant differences (t=5.491 3, 5.512 5, P<0.01). The length of stay was (18.34±3.58) d in the experimental group, and (21.82 ± 4.23) d in the control group, and there was significant difference(t=3.439 6, P<0.05). The cure rate was 93.33%(28/30) in the experimental group, and 56.67%(17/30) in the control group, and there was significant difference(χ2=10.755 6, P<0.05). Conclusions The comprehensive nursing intervention can provide comprehensive refinement of the overall care in children and their families, promote physical and psychological rehabilitation of children with severe burns, and repair their family and social support systems. The comprehensive nursing intervention has far-reaching implications for the rehabilitation of children with severe burns.

OBJECTIVE: To explore the genetic basis for a patient diagnosed with tuberous sclerosis complex (TSC). METHODS: Peripheral blood samples of the patient and his parents were collected for the extraction of genomic DNA. Next generation sequencing (NGS) was carried out to detect potential variant, and the result was verified by Sanger sequencing. RESULTS: The patient was found to harbor a heterozygous c.1053delG (p.Glu352SerfsX10) frameshifting variant of the TSC2 gene. The same variant was not found in his unaffected parents and 100 unrelated healthy controls. Based on the American College of Medical Genetics and Genomics guidelines, the variant was predicted to be pathogenic (PVS1+PS2+PM2). CONCLUSION The novel c.1053delG (p.Glu352SerfsX10) frameshifting variant of the TSC2 gene probably underlay the TSC in this patient.
Genomics ; Heterozygote ; Humans ; Mutation ; Tuberous Sclerosis/genetics* ; Tuberous Sclerosis Complex 2 Protein/genetics*

Objective To analyze histopathological and clinical features of dermatofibroma,and to explore the relationship between them.Methods Clinical and histopathological data were collected from 150 patients with histopathologically confirmed dermatofibroma in Department of Pathology,Shanghai Skin Disease Hospital from September 2017 to August 2018,and analyzed retrospectively.Results Among the 150 patients,65 were males,and 85 were females.Their age was 42 ± 13.8 years,and the course of disease ranged from 3 months to 30 years.Some of the patients had concomitant symptoms,mainly manifesting as itching,some had spontaneous pain and mild tenderness,and 18 patients had a history of injury,insect bite or infection at lesion sites.Skin lesions mainly occurred on the extremities (107 cases,71.3％),and most were solitary (105 cases,70％).Before pathological examinations,102 cases were clinically diagnosed as dermatofibroma,16 as epidermoid cyst,13 as pigmented nevus,3 as keloid,12 as skin mass,1 as malignant melanoma,1 as xanthogranuloma,1 as prurigo nodularis,and 1 as neurofibroma.Among 169 hematoxylin and eosin (HE)-stained sections,25 (14.8％) appeared to be consistent with aneurysmal dermatofibroma,66 (39.1％)with cellular dermatofibroma,36 (21.3％) with sclerosing dermatofibroma,and 22 (13.0％)with epithelioid dermatofibroma.Coexistence of two or more subtypes could be seen in 12 sections.There were also a few new variants,such as dermatofibroma with hyperplastic sweat duct (1 case),deep dermatofibroma (3 cases),dermatofibroma with epithelioid cells intermingled with hyperplastic collagen (1 case).The duration of aneurysmal dermatofibroma varied from 7 months to 30 years,and most manifested as skin masses on the lower extremities.A relatively short course of disease was observed in patients with cellular dermatofibroma,who often visited a hospital several months after the onset,and cellular dermatofibroma was commonly observed on the extremities and frequently accompanied with itching and pain.The duration of sclerosing or atrophic dermatofibroma was usually long for years or decades,and it commonly occurred on the upper limbs without concomitant symptoms.Epithelioid dermatofibroma of varied durations had various clinical manifestations,frequently occurred on the lower limbs without concomitant symptoms.Conclusions The clinical and pathological manifestations of dermatofibroma are diverse.Different dermatofibroma lesions can share similar typical histopathological manifestations,and atypical pathological features can interfere with the diagnosis of dermatofibroma.

Objective:To analyze clinical phenotypes and pathogenic mutations of a patient with classic tuberous sclerosis complex.Methods:Clinical data was collected from a patient with classic tuberous sclerosis complex. Next-generation sequencing was performed to screen pathogenic gene variants, and Sanger sequencing to verify the mutations. Minigene plasmids were constructed and transfected into the human renal epithelial cell line 293T, and RNA was extracted for transcriptional analysis.Results:The patient clinically presented with recurrent epileptic seizures, facial angiofibroma, periungual fibroma, pulmonary lymphangioleiomyomatosis, renal angiomyolipoma and multiple osteosclerosis. Next-generation sequencing revealed a suspected pathogenic variant in the TSC2 gene in the patient. Sanger sequencing identified a heterozygous mutation c.336_336+15delGGTAAGGCCCAGGGCG in exon 4 of the TSC2 gene in the patient, but not in his parents or 100 unrelated healthy controls. Moreover, this mutation had not been previously reported. The minigene experiment showed changed mRNA sequence of the TSC2 gene in this patient with loss of the authentic splice site in exon 4 and insertion of a 74-bp intron, which shifted the splice site 90 bp downstream (r.336delins336+16_336+90) .Conclusion:The novel heterozygous mutation c.336_336+15delGGTAAGGCCCAGGGCG in exon 4 of the TSC2 gene can lead to aberrant splicing, and may contribute to tuberous sclerosis complex in this patient.

Objective:To evaluate the efficacy and safety of transepithelial photorefractive keratectomy (Trans-PRK) combined with accelerated corneal cross-linking (CXL) for refractive error in patients with thin or irregular corneas, excluding keratoconus.Methods:An observational case series study was performed.Fifty-five right eyes of 55 myopic patients diagnosed with thin or irregular corneas, who underwent Trans-PRK combined with prophylactic CXL surgery, were included at Baotou Chaoju Eye Ophthalmic Hospital from August 2017 to July 2018.Uncorrected distance visual acuity (UDVA) of the operated eye was measured using international standard visual acuity charts, and refractive diopters were measured by computer and comprehensive refraction before surgery and at 1 week, 1, 3, 6, and 12 months after surgery.Corneal morphology was assessed with the Pentacam anterior segment analyzer before surgery and at 3, 6, and 12 months after surgery.Intraocular pressure (IOP) was measured with a non-contact tonometer before surgery and at 1, 3, 6, and 12 months after surgery.The incidence of postoperative complications was recorded.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Baotou Chaoju Ophthalmic Hospital (No.btcj-u-1). Written informed consent was obtained from each subject.Results:Preoperative, 1-week, 1-, 3-, 6-, and 12-month postoperative UDVA (LogMAR) were 0.52(0.55, 0.78), 0.22(0.12, 0.17), 0.10(0.04, 0.07), 0.00(-0.04, -0.16), -0.08(-0.05, -0.03) and -0.08(-0.06, -0.04), respectively, showing a statistically significant overall difference ( Z=249.44, P<0.001). UDVA at each postoperative time point was improved compared to preoperative, and UDVA at 3, 6, and 12 months postoperatively was significantly improved compared to 1 week and 1 month postoperatively (all at P<0.001). The spherical diopter at each postoperative time point decreased significantly compared to preoperative, with the spherical diopter at 1, 3, 6, and 12 months postoperatively being lower than that at 1 week postoperatively, and the 12-month postoperative spherical diopter being lower than that at 3 and 6 months postoperatively, showing statistically significant differences (all at P<0.001). The cylindrical degree at 1, 3, 6, and 12 months postoperatively was lower than that at preoperative and 1 week postoperatively, with statistically significant differences (all at P<0.05). After the operation, the spherical equivalent of the operated eye gradually decreased with time, tending toward emmetropia.The spherical equivalent at each postoperative time point decreased compared to preoperative, with the spherical equivalent at 1, 3, 6, and 12 months postoperatively being lower than that at 1 week postoperatively, and the spherical equivalent at 12 months postoperatively being lower than that at 3 and 6 months postoperatively, showing statistically significant differences (all at P<0.001). The corneal K1 and K2 values at 3, 6, and 12 months postoperatively were significantly lower than preoperatively (all at P<0.001), and the corneal K1 and K2 values at 3 months postoperatively tended to stabilize.The IOP of the operated eye at 3, 6, and 12 months postoperatively was significantly lower than preoperatively, and the IOP at 6 and 12 months postoperatively was lower than that at 1 and 3 months postoperatively, with statistically significant differences (all at P<0.001). One eye developed grade 0.5 corneal haze at 1 week postoperatively, which spontaneously resolved to transparency at 1 month postoperatively. Conclusions:Trans-PRK combined with accelerated CXL has good efficacy, stability and safety for refractive error patients with thin or irregular corneas, except for keratoconus.

To evaluate the clinical significance of nerve growth factor（NGF） in patients with cerebral small vessel disease（CVSD） and explore the relationship between NGF and the pathogenesis of depression after CVSD（CVSD-D）. Methods 80 Patients with first-onset CSVD were recruited. The CSVD were confirmed by MRI. Forty age and gender matched healthy people were selected as the control group. One month later，the diagnosis of post-stroke depression（PSD） was performed according to the American Classification and Diagnostic Criteria for Mental Disorders（DSM-IV）. Depressive symptoms are scored according to the 17-item Hamilton Depression Scale（HAMD-17 items）. The cubital venous blood was drawn within 24 hours after the depression scale was scored，and the serum NGF was detected by enzyme-linked immunosorbent assay（ELISA）. Results The serum NGF in patients with csvd were higher than those of normal patients，But in the CSVD patients，the serum NGF in patients with CSVD-D was lower than those of non CSVD-D group. The difference was statistically significant. Correlation analysis showed that there is no correlation between serum NGF and HAMD score（r=02096，P=0.3752）. Conclusion The occurrence of depression after cerebrovascular disease is closely related to the serum NGF level. But the degree of depression has no correlation with the NGF level.

Objective:To explore the effect of restrictive perineal incision evaluation combined with prone flexion delivery on reducing the perineal incision rate.Methods:The convenient sampling method was used to select 440 primiparas who delivered naturally in Wuxi People's Hospital Affiliated to Nanjing Medical University from June to November 2020 as research objects. According to the random number table method, the primiparas were divided into the observation group and the control group, with 220 cases in each group. The observation group was scored item by item according to the Restrictive Perineotomy Assessment Scale, and the indications of perineotomy were strictly performed according to the final score. The control group judged whether to undergo perineotomy according to subjective experience. The prone flexion delivery was used in both groups and the delivery outcomes of the two groups were compared.Results:The perineal incision rate of the observation group was lower than that of the control group, and the difference was statistically significant ( P<0.01) . The perineal laceration degree of the observation group was less than that of the control group, and the difference was statistically significant ( P< 0.05) . The rate of poor perineal wound healing in the observation group was lower than that in the control group, and the difference was statistically significant ( P<0.05) . The degree of postpartum perineal pain in the observation group was less than that in the control group, and the difference was statistically significant ( P<0.01) . There was no statistically significant difference in the incidence of neonatal asphyxia between the two groups ( P>0.05) . There was no neonatal injury in the two groups. Conclusions:Construction and application of Restrictive Perineotomy Assessment Scale avoid midwives to judge based on subjective experience and improve the accuracy of perineotomy assessment. The combination with prone flexion delivery can further reduce the rate of perineotomy and poor healing rate of postpartum perineal wound and reduce postpartum perineal pain, which is an effective method to improve postpartum perineal outcome and ensure the safety of mother and children.