Objective:To further optimize the nutritional support program for elderly neurocritical patients, analyze the changes and influencing factors of serum albumin levels in elderly neurocritical patients after surgery, and explore the impact on prognosis.Methods:A retrospective cohort study was conducted to investigate the medical records of neurosurgical intensive patients in Shanghai Tenth People's Hospital from May 2015 to September 2019. All the patients were treated for at least 5 days after operation. Patients with severe liver and kidney dysfunction, mental illness and incomplete medical records were not included in the study. Patients aged ≥65 years were included as the experimental group and patients aged <65 years served as the control group. Generalized estimating equation and other models were used to analyze the difference in the trend of albumin level and its influencing factors between the two groups. Automated machine learning was used to explore the influence of factors such as albumin level, nutrition mode, age, sex, diagnosis, preoperative consciousness level on the prognosis of patients.Results:A total of 284 patients were included in the study, including 85 in the experimental group and 199 in the control group. There were no significant differences in baseline data such as sex, age, and proportion of nutritional patterns between the two groups. Generalized estimating equation and generalized mixed additive equation showed that albumin level in both groups reached the minimum on the 3rd day after operation, and the average level in the experimental group was significantly lower than that in the control group ( P=0.03). Cerebral hemorrhage, emergency surgery, and low preoperative albumin were risk factors ( P<0.05); intravenous supplementation of human albumin might have an adverse effect on the experimental group ( P = 0.047). Machine learning models suggested that the higher the serum albumin level at discharge, the better the prognosis at 1 month after surgery. The area under ROC curve was 0.906, the specificity was 93.7%, and the sensitivity was 67.0%. Conclusions:The changes of albumin level in elderly patients with neurological critical illness after surgery are basically the same as those in young patients, with the lowest albumin level on the 3rd day after surgery, and the albumin levels even lower on the 3rd day after surgery in patients with cerebral hemorrhage, emergency surgery and preoperative low albumin. The higher the albumin level at discharge, the better the prognosis of patients may be.

Objective To measure the expressions of Toll like receptors (TLRs) in lesions of condyloma acuminatum (CA). Methods Tissue samples were resected from lesions of 25 patients with CA and normal skin of 16 human controls. Frozen sections were prepared and the expressions of TLRs 1 - 10 were measured by immunohistochemical staining. Results In normal skin samples, there was a strong expression of TLR 6 and 9, medium expression of TLR 1, 3, 10 and weak expression of TLR 2, 4, 5, 7, 8 in the stratum corneum and stratum granulosum, but a weak or negative expression of TLRs in the stratum spinosum and stratum basale. In contrast, a strong expression of TLR 1, 3, 6, 9, medium expression of TLR 4, 8, 10 and weak expression of TLR 2, 5, 7 were detected in keratinocytes of almost all epidermal layers except for stratum basale in CA lesions. Conclusions The expressions of TLRs, especially TLR 1, 3, 6, 9, are markedly enhanced in proliferated keratinocytes in CA lesions compared with normal skin, which is likely to be associated with anti-viral immunity triggered by HPV infection.

Objective:To explore the clinical effect of negative pressure wound therapy and to discuss the ethi-cal issues in this therapy and seek a solution. Methods:Sixty patients with negative pressure wound therapy in our hospital were invited to participate in this study. Patients were all with deep pressure ulcers or diabetic foot, of which the Wagner Scores were graded as three or four. They were divided into two groups randomly. The subjects in control group received standard treatment, while those in experimental group were treated by self -designed closed negative pressure drainage settings. The wound healing time, the cost of the treatment and comfort were compared between the two groups. Results:The cost of wound care in the experimental group was less than those in the control group (P<0. 001). There was no significant difference in comfort between two groups (P>0. 05). But the wound healing time was longer in the experimental group than those in the control group. Conclusion:Self-designed closed negative pressure settings can decrease the cost of patient care with the equal patient comfort and acceptance as the standard negative pressure, however, the wound healing time has been extended. So before using negative pressure technique, the medical staff should comprehensively analyze, be guided by medical ethics and choose the appropriate treatment for different patients.

Abstract: Objective To investigate the occurrence of multidrug-resistance among tuberculosis patients in Hainan Province from 2014 to 2020 and to analyze the influencing factors, aiming to provide reference for formulating drug-resistant tuberculosis control strategies in this region. Methods This study collected sputum samples from the patients with pulmonary tuberculosis admitted to the Second Affiliated Hospital of Hainan Medical University from 2014 to 2020, and performed isolation and identification of Mycobacterium tuberculosis and drug susceptibility testing. After the strains were identified as positive, drug sensitivity tests were conducted, and multi-drug resistant patients were found. Clinical data was retrospectively collected, and chi-square test and unconditioned logistic regression were used to analyze the influencing factors of multidrug resistance. Results A total of 2 672 patients underwent sputum culture, strain identification, and drug susceptibility testing in TB designated hospitals in Hainan Province from January 1, 2014 to December 31, 2020. Among them, 1 942 patients with available drug susceptibility test results and complete clinical data were enrolled, among which 398 cases with drug-resistant TB were included in the case group, and 1 544 cases without drug resistance were included in the control group. Multivariate logistic regression analysis showed that farmers, rural residence, treatment history of retreatment, irregular medication history, number of pulmonary cavities ≥3, and BMI<18.5 were independent risk factors for MDR-TB. The risk of MDR-TB in farmers was higher than that in non-farmers (OR=1.542, 95%CI: 1.150-2.020); patients living in rural areas had a higher risk of multidrug resistance than those living in urban areas (OR=1.445, 95%CI: 1.095-1.907); the risk of MDR in the retreatment patients was higher than that in the initial treatment patients (OR=5.616, 95%CI: 4.250-7.421); the risk of multi-drug resistance in patients with irregular medication was higher than that in patients with regular medication (OR=2.665, 95%CI: 2.012-3.531); the risk of multidrug resistance in patients with pulmonary cavity number ≥3 was higher than that in patients with pulmonary cavity number <3 (OR=5.040, 95%CI: 3.768-6.740); compared with patients with BMI<18.5, patients with BMI=18.5-24.0 and BMI≥24.0 had a lower risk of multidrug resistance (OR=0.735, 95%CI: 0.555-0.975 and OR=0.447，95%CI:0.225-0.888, respectively). Conclusions Retreatment, farmer occupation, rural residence, irregular medication and low BMI may be the risk factors for multidrug resistance in Hainan Province.

The purpose of this study is to investigate the applicability of a natural swelling matrix derived from boat-fruited sterculia seed (SMS) as the propellant of osmotic pump tablets. The sugar components, static swelling, water uptake and viscosity of SMS were determined and compared with that of polythylene oxide (WSR-N10 and WSR-303). Both ribavirin and glipizide were used as water-soluble and water-insoluble model drugs. Then, the monolayer osmotic pump tablets of ribavirin and the bilayer osmotic pump tablets of glipizide were prepared using SMS as the osmotically active substance and propellant. SMS was mainly composed of rhamnose, arabinose, xylose and galactose and exhibited relatively high swelling ability. The area of the disintegrated matrix tablet was 20.1 times as that at initial after swelling for 600 s. SMS swelled rapidly and was fully swelled (0.5%) in aqueous solution with relative low viscosity (3.66 +/- 0.03) mPa x s at 25 degrees C. The monolayer osmotic pump tablets of ribavirin and the bilayer osmotic pump tablets of glipizide using SMS as propellant exhibited typical drug release features of osmotic pumps. In conclusion, the swelling matrix derived from boat-fruited sterculia seed, with low viscosity and high swelling, is a potential propellant in the application of osmotic pump tablets.

Objective To observe the levels of Foxp3+ CD+ CD25+ regulatory T cells in the peripheral blood of condyloma acuminatum (CA) patients and investigate their roles in the pathogenesis of CA. Methods The peripheral blood was collected from 30 CA patients (including 15 with relapsing and 15 with first onset) and 20 healthy controls. Peripheral blood mononuclear ceils (PBMC) were isolated and stained with anti-human CD4-PE-Cy5 and anti-human CD25-fluorescein isothiocyanate (FITC) monoclonal antibodies on cell membrane, followed by intraeellular staining with anti-human Foxp3-PE. The percentage of Foxp3+ CD4+-CD25+ regulatory T cells was detected by three-color flow cytometry. Comparison between groups was done by ANOVA test. Results The percentages of Foxp3+ CD4+ CD25+ regulatory T cells among total CD4 + T cells in CA patients and relapsing CA group were (3.4 ± 1.0) % and (4.7 ±+ 1.2) %, respectively, which were both significantly higher than that in healthy control group [(1.2±0. 5)%, P<0.01]. Furthermore, that in first onset CA group was (2. 1 ± 1.0) %, which was higher than that in healthy control group, but without statistical significance; but that in relapsing CA group was significantly higher than that in first onset group (P<0.05). Conclusions The number of Foxp3+ CD4+ CD25+ regulatory T cells increases in the peripheral blood of CA patients. The disorder of cellular immunity may be involved in the immunopathogenesis of CA.

Objective To screen and identify the predicted epitopes of synthesized HLA-A*0201restricted CTL derived from HPVll E7 antigen.Methods Five HPVll E7 CTL epitope peptides and terramers consisting of HLA-A*0201 were selected by way of computer and synthesized by Sanquin company,including HPVllE7 7-15(TLKDIVLDL),15-23(LQPPDPVGL),47-55(PLTQHYQIL),81-89(DLLLGTLNI)and 82-90(LLLGTLNIV).These peptides binding to human peripheral blood-derived DCs were tested for their ability to activate T cells isolated from peripheral blood lymphocytes of HLA-A*0201 healthy individuals.the number of specific tetramer+CD8+T cells by flow cytometry,the level of the section of IFN-γ by ELISA,and the ability of the CTL to kill the target cells were observed.Results The immature DCs could be fully activated by all the five HPV11 E7 peptides.Peptide-loaded mature DCs were able to stimulate the epitope-specific T cells responses in vitro.An increased frequency(P<0.05)of T ceils specific for the E7 7-15 epitope compared to other epitopes of HPV11E7.The epitope-specific CTL of E7 7-15 induced by the activated DCs specifically killed HPV11E7 expressing 293 cell line,and in a ratio of 50:1,the specific cytolytic activity was the strongest than the others(P<0.05).Conclusion DCs loaded with HPV11 E7 7-15(TLKDIVLDL)peptide can induce highly effective and specific ectogenic processed epitopespecific CTL responses in vitro.This peptide may be the candidate for development of CTL based vaccine in the treatment of HPV infeetions.

Objective To construct a prokaryotic plasmid to express the testis development related gene 1 (TDRG1) recombinant protein and obtain anti-TDRG1 mAb by immunizing mice, and to identify the biological properties of the mAb. Methods The coding sequence of TDRG1 was amplified by RT-PCR from normal human testis tissue and cloned into the vector pET21, and then was expressed in the E. coli BL 21(DE3) to get TDRG1 recombinant protein. The purified TDRG1 recombinant protein was injected to immunize the BALB/C mice to develop anti-TDRG1 mAb. Splenocytes of the immunized mice were collected and fused with the mouse myeloma cell line Sp2/0 cells. The hybridoma cells that secreted anti-TDRG1 mAb were subcloned with limited dilution. Enzyme-linked immunosorbent assay (ELISA) was used to evaluate titers and subtypes of mAb. Western blot and immunohistochemistry were used to detect specificity of mAb.Results The prokaryotic plasmid expressing the recombinant protein was constructed, and the TDRG1 recombinant protein was expressed and purified. Two hybridoma cell lines that secreted anti-TDRG1 mAb were obtained. The titer of the mAb in ascites was 1∶1.6×10~6, and the subtype of the mAb was IgG_1. Westem blot and immunohistochemistry analysis indicated the mAb showed specific combination with TDRG1 protein in human testes.Conclusion The TDRG1 recombinant protein is highly purified and has strong antigenicity. The anti-TDRG1 mAb is produced successfully.

Objective:To construct a p53-fused dual luciferase reporter and to test whether this reporter can mimic wild-type p53 activities in a high-throughput screen.Methods:A restriction endonuclease site was added to each terminus and the stop codon of the wild-type full-length p53 open reading frame (ORF) was removed by PCR. A restriction endonuclease site was added to each terminus and the start codon of the ifrelfy luciferase ORF was removed by PCR. The two modified ORFs were inserted upstream of the IRES-induced renilla luciferase ORF in a CMV-derived vector. hTe p53 fusion protein was expressed in cells to test its MDM2-mediated degradation, subcellular localization, and induction of p53-responsive promoter.
Results:hTe p53-fused dual luciferase reporter was successfully constructed. Atfer transfection into the host cells, the reporter expressing the p53 fusion protein that was degraded by oncoprotein MDM2, was mainly located inside the nucleus, and induced the p53-responsive promoter, respectively.
Conclusion:hTe p53-fused dual luciferase reporter (p53FL/IRES/RL) can identify modulators of P53 protein level in a high-throughput screen of genetic or chemical libraries.

AIM:To explore the differential expression of interleukin-19 (IL-19) and its receptors (IL-20R1/IL-20R2) in the samples of chronic rhinosinusitis (CRS) with or without nasal polyps (CRSwNP and CRSsNP), and to investigate the correlation of IL-19 and its receptors with tissue remodeling factors, including matrix metalloproteinase (MMP)-2, MMP-9 and tissue inhibitor of metalloproteinase (TIMP)-1 in CRS.METHODS:The polyps from CRSwNP patients (n=30), the sinus mucosa from CRSsNP patients (n=15) and the inferior turbinal mucosa of nasal septum de-viation patients (n=15) were collected.The expression of IL-19 and its receptors (IL-20R1/IL-20R2) in each group was detected by real-time RT-PCR and immunohistochemistry.The expression of tissue remodeling factors MMP-2, MMP-9 and TIMP-1 in different groups was detected by real-time RT-PCR and ELISA.RESULTS:Increased mRNA and protein expression levels of IL-19, IL-20R1, IL-20R2 and MMP-9 were observed in CRSwNP group compared with CRSsNP and control group (P<0.05), while elevated expression level of TIMP-1 was observed in CRSsNP group compared with CRSwNP and control group (P<0.05).The relative expression of MMP-2 among the 3 groups showed no obvious difference.The expression of IL-19 and its receptors showed significantly positive correlations with MMP-9 in CRSwNP (P<0.05).No significant correlation between IL-19 and/or its receptors with TIMP-1 in CRSwNP group was observed.CONCLUSION:The high expression levels of IL-19, IL-20R1 and IL-20R2 and their positive correlations with MMP-9 in CRSwNP indica-te that IL-19 and its receptors may be involved in the tissue remodeling of chronic rhinosinusitis.