Objective To study the expressions of Toll-like receptor 9 protein (TLR9) in peripheral B and T lymphocytes in newly diagnosed, untreated patients with systemic lupus erythematosus (SLE) and their relationship with clinical parameters. Methods Blood samples were obtained from 35 newly diag-nosed, untreated patients with SLE and 16 healthy human controls. B, T lymphocytes and TLR9 protein were labeled with fluorescent antibodies, and the expressions of TLR9 protein were detected by flow cytometry in peripheral B and T lymphocytes. The relationship between TLR9 expression and clinical parameters was assessed. Results The proportions of B and T lymphocytes expressing TLR9 in newly diagnosed, untreated patients were (53.94±17.95)% and (49.33 ± 23.30)%, respectively, compared to (29.40 ± 10.54)% and (29.18 ± 14.78)%, respectively, in healthy controls (t = 6.11,3.73, respectively, both P < 0.01). Additionally,the proportion of B lymphocytes expressing TLR9 correlated negatively with SLE disease activity index (SLEDAI)(r = -0.39, P < 0.05), but positively with the level of serum IgA antibody (r = 0.74, P < 0.01).Condnsions The expression of TLR9 is elevated in peripheral T and B lymphocytes from patients with newly diagnosed, untreated SLE, and the proportion of TLR9-expressing B lymphocytes negatively correlates with SLEDAI, but positively correlates with the serum level of IgA antibody.

Purpose To assess left ventricular diastolic function in the patients with essentialhypertension. Methods 25 normotensives,93 hypertensiives without hypertrophy (NLVH) and 47 withhypertrophy (LVH)(LVMI, left ventricular mass index ＞ 124 g/m2 in males and 110 g/m2 in females)underwent 2DE and Doppler echocardiography. Results The ratio of early to late peak velocity (E/A)was 1.21 ± 0.24 in normotensives and 1.03 ± 0.23 in NLVH patients( P ＜ 0.01 ); reversed pulmonaryvenous flow at atrial systole PA wave was (32.7 ± 7.5 ) vs (38.9 ± 8.7) cm/s (P ＜ 0.01 ) in normotensivesand in NLVH patients respectively. The isovolumic relaxation time (IVRT) and left atrial dimension (LAD)have significant differences in three groups ( P ＜ 0. 05 - 0. 001 ). EF value was similar betweennormotensives and NLVH patients, but it was lower in LVH patients ( P ＜ 0.01 ). Conclusions E/Aratio, PA wawe, Pai, IVRT and LAD are sensitive parameters indicating mild diastolic left ventriculardysfunction. Systolic left ventricular function has no change in NLVH patients.

Purpose To investigate the expression pattern and c1inicopatho1ogica1 significance of D2-40 in craniopharyngioma( CP). Methods Immunohistochemica1 method was used to assess D2-40 expression in 126 cases of craniopharyngioma. Statistic software was used to ana1yze the corre1ation between D2-40 expression and c1inicopatho1ogica1 features. Results The overa11 positive rate of D2-40 expression in 126 craniopharyngioma was 87. 30%. The rate of +, and  were 44. 44%( 56/126 ),37. 30%( 47/126 )and 5. 56%(7/126)respective1y. In adamantinomatous CPs,D2-40 expression was observed in epithe1ia1 components corresponding to the stratum intermedium,whi1e in papi11ary CPs,it was immuno-positive in basa1 ce11s. With the increasing of the existence of inf1amma-tion in tumor,D2-40 expression was up-regu1ated. Tumor ce11s were over-expressed for D2-40 in basa1 ce11s and stratum intermedium ce11s of the invasive frontier. D2-40 expression was higher in the invasive craniopharyngioma than in the non-invasive craniopharyngio-ma. In addition,D2-40 expression was higher in the recurrent craniopharyngioma than in the non-recurrent craniopharyngioma. There were no significant re1ationship between D2-40 expression and gender,and histo1ogica1 c1assification of tumors. Conclusions D2-40 expression is associated with the deve1opment of craniopharyngioma,which can 1ead to enhance the tumor ce11 invasion,and may be re-1ated to inf1ammation-re1ated mechanisms. D2-40 expression may be one of the recurrence factors in patients with craniopharyngioma.

Objective To establish normally immortalized porcine hepatocyte lines by ectopic expression of simian virus 40 large T (SV40LT) antigen and the human telomerase reverse transcriptase(hTERT). Methods Primary porcine Hepatoeyte cells were transfeeted with recombinant retrovirus containing SV40LT or hTERT respectively. Subsequently drug resistant cell clones were screened and expanded for further studies. Immortalized porcine hepatocyte was confirmed by examination. Results The morphological phenotype of the transfected cells was similar to the primary porcine hepatocyte. One clone, HepLP, has been maintained in cultue for half year, and expanded by more than 60 passages. SV40 LT and hTERT could be detected in transfected porcine hepatocyte. Pig albumin mRNA was also detected by RT-PCR. No tumor formation occurred when HepLP cells were injected into Balb/c nude mice. Conclusions The immortalized, nontumorigenic, porcine hepatoeytes maintained the properties of porcine primary hepatocytes such as the albumin secretion. This generation of immortalized porcine hepatocyte may be helpful for bioartifical liver support system, hepatocytes transplantation, drug/toxicological studies, and liver biologic studies.

OBJECTIVETo test the effects of 7 virus-encoded RNA silencing suppressors (RSSs) for enhancement of a plant virus-based vector system-mediated heterologous expression of green fluorescence protein (GFP) in Nicotiana benthamiana.

METHODSSeven transient expression vectors for the 7 RSSs were constructed and co-inoculated on the leaves of Nicotiana benthamiana with PVXdt-GFP vector, a novel Potato virus X-based plant expression vector, through agroinfiltration. The protein and mRNA expression levels of the reporter gene GFP in the co-inoculated Nicotiana leaves were examined by Western blotting, ELISA and RT-qPCR to assess the effect of the RSSs for GFP expression enhancement.

RESULTSThe 7 RSSs differed in the degree and duration of enhancement of heterologous GFP expression, and the p19 protein of Tomato bushy stunt virus (TBSV) induced the highest expression of GFP. African cassava mosaic virus AC2 protein and Rice yellow mettle virus P1 protein produced no obvious enhancement GFP expression.

CONCLUSIONTransient co-expression of RSSs suppresses host silencing response to allow high-level and long-term expression of heterologous genes in plant, but the optimal RSS has to be identified for each plant virus-based expression vector system.

Genetic Vectors ; Green Fluorescent Proteins ; genetics ; Plant Viruses ; genetics ; Plants, Genetically Modified ; genetics ; metabolism ; Potexvirus ; genetics ; RNA Interference ; Tobacco ; genetics ; metabolism

Objective To establish immortalized human hepatocyte lines for studies of bioartificial liver,hepatocyte transplantation,and drug metabolism in vitro.Methods Primary human hepatocytes were isolated by 4-step perfusion technique with collagenase and transfected with recombinant retrovirus containing Simian virus 40 large T antigen(SV40 LT).Subsequently,immortalized human hepatocytes were evaluated by analysis of gene expression and functional characteristics in vitro.Results Two immortalized human hepatocyte lines,HepLi2 and HepLi3,were obtained after primary human hepatocytes being infected by SV40 LT containing recombinant retrovirus for 3-4 weeks.The immortalized human hepatocytes showed classical appearance of hepatocyte observed by phase contrast microscope.The protein expression of SV40 LT in HepLi2 and HepLi3 cells were detected by Western blotting.The mRNA expressions of albumin(Alb),glutathione S-transferase(GST-p),human blood coagulation factor X(HBCF-X)and β-actin in HepLi2 and HepLi3 cells were detected by reverse transcription polymerase chain reaction(RT-PCR),and the mRNA expressions of cytochrome(CY)450 subtypes(CYP3A5,CYP2E1,CYP2C8-19 and CYP3A4)in HepLi2 and HepLi3 cells were also observed by RT-PCR.Levels of alanine transaminase (ALT),lactate dehydrogenase(LDH)and Alb were detected in the supernatant of immortalized human hepatoeyte culture.Conclusions The immortalized human hepatocyte lines have the biological characteristics of primary human hepatocytes and have the CYP450 functions of hepatocytes,which may be heIDful for the studies of bioartificial liver,heoatocvte transplantation and drug metabolism in vitro.

In the COVID-19 epidemic prevention and control work, not only the safety management of in-hospital organ transplantation is facing severe challenges, but also a large number of patients at home after transplantation are in immunosuppressive state, and professional protection management is urgently needed. Since the outbreak of the epidemic, Shulan(Hangzhou)hospital fully identified the risks of transplant patients, established a special team, and formulated and implemented the control management plan for organ transplant patients based on the crisis leadership model. The control management plan has achieved staged results.From January 15 to March 2, 2020, 29 liver transplants and 31 kidney transplants were performed in our hospital. The remote health education of 1 002 patients after liver and kidney transplantation was completed. The goal of " zero infection" was achieved, and the protection management quality indicators were ideal.

Objective:To investigate the changes of paraspinal muscles in patients with degenerative lumbar scoliosis (DLS) and its correlation with lumbar kyphosis.Methods:The clinical data of 67 female patients with degenerative lumbar scoliosis, with an average of 65.4±5.6 years old (rang 52-83 years old), were retrospectively analyzed. There were 35 patients of DLS with lumbar degenerative kyphosis (LDK) in the DLS+LDK group, with an average of 64.60±5.40 years old (rang 52-75 years old), and 32 patients of lumbar scoliosis without lumbar kyphosis in the DLS group, with an average of 66.22±5.8 years old (rang 55-83 years old). The cross-sectional area (CSA) and the percentage of fat infiltration area (FIA%) of erector spinae and multifidus muscles of the 5 intervertebral disc levels (from L 1-2 to L 5S 1) were measured by MRI using Image J software (ver. 1.51 k, National Institutes of Health, USA). The curve direction, Cobb angle, sagittal vertical axis (SVA), thoracic kyphosis (TK), thoracolumbar kyphosis (TLK), lumbar lordosis (LL), pelvic incidence (PI), pelvic tilt (PT) and sacral slope (SS) were evaluated and recordedin both groups using an anteroposterior radiograph in the standing position, and the correlation between the changes of paraspinal muscles and these factors was analyzed. Results:The TLK, LL, and SVA values of the DLS+LDK group (11.85°±7.89°, -9.35°±8.70° and 70.16±76.94 mm) were higher than those of the DLS group (7.47°±5.06°, -26.46°±10.26° and 39.45±38.18mm) ( t=2.73, P=0.008; t=7.38, P<0.001; t=2.10, P=0.041). The TK, PI, and SS values of the DLS+LDK group (16.36°±13.52°, 42.49°±11.70° and 11.89°±10.03°) were lower than those of the DLS group (23.60°±10.23°, 49.38°±11.92° and 21.21°±8.28°) ( t=2.45, P=0.017; t=2.38, P=0.020; t=4.13, P<0.001). The differences of Cobb and PT were not statistically significant between the two groups. The cross-sectional areas of L 1-2, L 2-3, L 3-4 intervertebral disc levels of erector spinae of the DLS+LDK group (1 328.36±339.16 mm 2, 1 331.98±305.76 mm 2 and 12 53.58±275.86 mm 2) were lower than those of the DLS group (1 564.16±312.68 mm 2, 1 574.80±325.92 mm 2 and 1 427.18±278.82 mm 2) ( t=0.40, P=0.004; t=0.81, P=0.002; t=0.306, P=0.013). The cross-sectional areas of L 1-2, L 2-3, L 3-4, L 4-5 intervertebral disc levels of multifidus muscles of the DLS+LDK group (225.07±59.80 mm 2, 228.38±87.44 mm 2, 436.40±117.99 mm 2 and 666.55±184.13 mm 2) were lower than those of the DLS group (264.28±44.27 mm 2, 384.85±75.52 mm 2, 576.10±109.92 mm 2 and 801.52±145.83 mm 2) ( t=0.21, P=0.004; t=0.42, P<0.001; t=0.52, P<0.001; t=0.37, P=0.002). The differences of FIA% of erector spinae and multifidus muscles at all lumbar spine levels were not statistically significant between the two groups. The cross-sectional areas of L 1-2, L 2-3, L 3-4 intervertebral disc levels of erector spinae and L 1-2, L 2-3, L 3-4, L 4-5 intervertebral disc levels of multifidus muscles of the two groups were negatively correlated with LL values ( r=-0.37, P=0.002; r=-0.34, P=0.005; r=-0.21, P=0.049; r=-0.34, P=0.005; r=-0.61, P<0.001; r=-0.65, P<0.001; r=-0.55, P<0.001), and positively correlated with SS ( r=0.42, P<0.001; r=0.37, P=0.002; r=0.27, P=0.027; r=0.38, P=0.001; r=0.53, P<0.001; r=0.46, P<0.001; r=0.42, P<0.001). The cross-sectional areas of L 3-4 intervertebral disc levels of erector spinae and L 1-2, L 2-3 intervertebral disc levels of multifidus muscles of the two groups were positively correlated with PI ( r=0.25, P=0.039; r=0.33, P=0.006; r=0.35, P=0.004). There was no correlation between the FIA% of erector spinae and multifidus muscles at all lumbar spine levels and the sagittal and pelvic parameters in both groups. Conclusion:Paravertebral muscle atrophy is more obvious in patients with degenerative lumbar scoliosis with lumbar kyphosis, which may be related to the reduce of lumbar lordosis and sacral slope. Patients with lumbar scoliosis with a smaller PI are more likely to experience paravertebral atrophy and increased loss of lumbar lordosis, and ultimately leading to lumbar kyphosis.

Objective:To investigate the relationship between health promoting behavior and frailty in elderly patients with proliferative diabetic retinopathy(PDR),and the mediating effects of self-efficacy and loneliness.Methods:In an eye hospital of Anhui Province,214 elderly patients with PDR were selected from May 2021 to Nov 2022 by convenient sampling method.The Frailty Scale,Chinese Version of Health-Promoting Lifestyle Profile-Ⅱ,Self-Efficacy Scale and Simplified Loneliness Scale were used in this survey.Bootstrap method of Process software was used to analyze the mediating effect of self-efficacy on the relationship between health promoting behaviors and frailty in elderly PDR patients and the moderating effect of loneliness on the relationship between self-efficacy and frailty.Results:A total of 220 questionnaires were distributed and 214 valid questionnaires were returned,with valid response rate of 97.27% .Moderated mediation effect analysis suggested that health-promoting behaviors negatively predicted frailty(β=-0.508,P＜0.01).Health promoting behaviors and self-efficacy had significant predictive effects on frailty(β=-0.191 and-0.433,P＜0.01),and health promoting behaviors also had a significant predictive effect on self-efficacy(β=0.063,P＜0.01).Self-efficacy played a partially mediating role between health promoting behaviors and frailty,and the mediating effect accounted for 14.76% of the total effect.The product term of loneliness and self-efficacy significantly predicted frailty(β=0.255,P＜0.01),the mediating effect of self-efficacy on frailty was moderated by loneliness.Conclusions:The health-promoting behaviors of elderly patients with PDR affect frailty through self-efficacy,and loneliness moderates the relationship between self-efficacy and frailty.The moderated mediation model is established.

Objective:To investigate the effect of eyebrow arch augmentation with personalized polyether ether ketone (PEEK) implant assisted by endoscope.Methods:From January 2019 to August 2021, 9 patients with low and flat eyebrow arch requiring eyebrow arch augmentation were treated in the plastic surgery department of the Friendship Plastic Surgery Hospital Affiliated to Nanjing Medical University. The cranial and maxillofacial CT scanning data were obtained before operation, the eyebrow arch elevation range and height were designed according to the patient's requirements, and personalized peek implants were manufactured. During the operation, a small temporal hairline incision was used to accurately separate the implantation cavity of individualized peek implants under the periosteum of the eyebrow arch implantation area with the aid of endoscope. Personalized PEEK implants were implanted under endoscope to make them fully fit with the eyebrow arch bone and fixed with titanium nails. Photos were taken and analyzed before and after operation.Results:All patients were followed up for 3 months to 2 years. Their facial expressions were normal, and there were no motor nerve injury, rejection and iatrogenic infection. The patients were satisfied with the postoperative effect. Compared with before operation, the low level of orbital bone in eyebrow area was improved, and the facial contour was three-dimensional and harmonious.Conclusions:The eyebrow arch augmentation with individualized peek implant assisted by endoscope is safe and effective. It can greatly reshape the upper contour of the face, deepen the upper eyelid fossa and improve the convex eye phenomenon, so as to achieve the cosmetic effect of enhancing the three-dimensional sense of the eyebrow orbital area, which is worthy of clinical application.