A vimentin-targeting oral compound with host-directed antiviral and anti-inflammatory actions addresses multiple features of COVID-19 and related diseases

Damage in COVID-19 results from both the SARS-CoV-2 virus and its triggered overreactive host immune responses. Therapeutic agents that focus solely on reducing viral load or hyperinflammation fail to provide satisfying outcomes in all cases. Although viral and cellular factors have been extensively profiled to identify potential anti-COVID targets, new drugs with significant efficacy remain to be developed. Here, we report the potent preclinical efficacy of ALD-R491, a vimentin-targeting small molecule compound, in treating COVID-19 through its host-directed antiviral and anti-inflammatory actions. We found that by altering the physical properties of vimentin filaments, ALD-491 affected general cellular processes as well as specific cellular functions relevant to SARS-CoV-2 infection. Specifically, ALD-R491 reduced endocytosis, endosomal trafficking, and exosomal release, thus impeding the entry and egress of the virus; increased the microcidal capacity of macrophages, thus facilitating the pathogen clearance; and enhanced the activity of regulatory T cells, therefore suppressing the overreactive immune responses. In cultured cells, ALD-R491 potently inhibited the SARS-CoV-2 spike protein and human ACE2-mediated pseudoviral infection. In aged mice with ongoing, productive SARS-CoV-2 infection, ALD-R491 reduced disease symptoms as well as lung damage. In rats, ALD-R491 also reduced bleomycin-induced lung injury and fibrosis. Our results indicate a unique mechanism and significant therapeutic potential for ALD-R491 against COVID-19. We anticipate that ALD-R491, an oral, fast-acting, and non-toxic agent targeting the cellular protein with multipart actions, will be convenient, safe, and broadly effective, regardless of viral mutations, for patients with early- or late-stage disease, post-COVID complications and other related diseases. IMPORTANCEWith the Delta variant currently fueling a resurgence of new infections in the fully-vaccinated population, developing an effective therapeutic drug is especially critical and urgent in fighting COVID-19. In contrast to the many efforts to repurpose existing drugs or address only one aspect of COVID-19, we are developing a novel agent with first-in-class mechanism-of-actions that address both the viral infection and the overactive immune system in the pathogenesis of the disease. Unlike virus-directed therapeutics that may lose efficacy due to viral mutations and immunosuppressants that require ideal timing to be effective, this agent, with its unique host-directed antiviral and anti-inflammatory actions, can work against all variants of the virus, be effective during all stages of the disease, and even resolve post-disease damage and complications. A further development of the compound will provide an important tool in the fight against COVID-19, its complications, as well as future outbreaks of new viruses.

Normocalcemic with elevated post-operative parathormone in primary hyperpara-thyroidism: 9 case reports and literature review / 北京大学学报(医学版)

OBJECTIVE@#To summarize and analyze the clinical characteristics of primary hyperpara-thyroidism (PHPT) with normocalcemic parathormone elevation (NPE) after surgical treatment, so as to improve the therapeutic ability and standardized post-operative follow-up of PHPT patients.@*METHODS@#Nine patients who were diagnosed with PHPT in the Department of Endocrinology of China-Japan Friendship Hospital from August 2017 to November 2019 were selected as the subjects. They all developed NPE within 6 months after surgical treatment. The clinical features and outcomes were collected and analyzed retrospectively, in addition, the related literature was reviewed.@*RESULTS@#Clinical features: among the 9 patients, 6 were middle-aged and elderly females and 3 were male. The main clinical manifestations were bone pain, kidney stones, nausea and fatigue except for one case of asymptomatic PHPT. Pre-operative examination showed high serum calcium [(3.33±0.48) mmol/L], low serum phosphorus [0.76 (0.74, 0.78) mmol/L], high 24-hour urinary calcium [8.1(7.8, 12.0) mmol/24 h], obviously elevated intact PTH [(546.1±257.7) ng/L], vitamin D deficiency [25-hydroxyvitamin D3 (21.0±5.7) nmol/L]. Serum levels of bone alkaline phosphatase [7 patients 41.3(38.6, 68.4) μg/L, 2 patients >90 μg/L] and N-terminal midcourse osteocalcin (>71.4 μg/L) were significantly elevated. The estimated glomerular filtration rate decreased in 2 patients. Imaging examination: 7 patients had osteoporosis. Renal calculi were found in 3 patients by renal ultrasound. Imaging examination of parathyroid glands found definite lesions in all the patients, including 2 cases of multiple lesions and 7 cases of single lesions.@*TREATMENT AND OUTCOME@#two patients underwent parathyroidectomy, while other patients were treated with microwave thermal ablation. PTH increased 1 month after therapy [(255.0±101.4) ng/L], and no recurrent lesions were found by parathyroid ultrasound. After combined treatment with cal-cium and vitamin D for six months, PTH decreased significantly and the level of serum calcium remained normal at anytime during the follow-up period.@*CONCLUSION@#The occurrence of postoperative NPE may be related to the higher pre-operative PTH, vitamin D deficiency and lower creatinine clearance. However, NPE may not predict recurrent hyperthyroidism or incomplete parathyroidectomy. Adequate calcium and vitamin D supplementation after surgery seems to be beneficial for patients with NPE. Post-operative follow-up of PHPT patients should be standardized to prevent and treat post-operative NPE.

Discharge and Contents of Exosome from Leukocyte-Reduced Apheresis Platelet Stored for Different Storage Time / 中国实验血液学杂志

OBJECTIVE@#To explore the effect of storage time on discharge and content of exosome from leukocyte-reduced apheresis platelets (LRA-Plt).@*METHODS@#Exosome (EXO) from LRA-Plt were acquired by ExoQuick, and its' morphology, immunological marker and particle size distribution were detected by transmission electron microscopy (TEM), Western blotting and dynamic light scattering (DLS), respectively. The changes in particle size distribution of EXO from LRA-Plt with different storage time were detected by DLS. The changes in content of protein and RNA of EXO from LRA-Plt with different storage time were detected by Nanodrop® ND-2000.@*RESULTS@#EXO from LRA-Plt was acquired successfully, which was characterized by cup-like shape, CD63/TSG101 enriched and Calnexin negative, and the particle size of which ranged from 30 to 200 nm. At early stored stage (stored for 1 day and 2 days), particle size of EXO from LRA-Plt was small and ranges from 30 to 40 nm. Meanwhile, the contents of protein and RNA were low. The particle size distribution, contents of protein and RNA of EXO from LRA-Plt were not significanty different ammg groups (P＞0.05). At middle-late stored stage (stored for 3, 4 and 5 days), the particle size of EXO from LRA-Plt was larger than that of early stored stage, which ranges was from 130 to 200 nm. Meanwhile, the contents of protein and RNA were higher than those of early stored stage. Particle size distribution, contents of protein and RNA of EXO from LRA-Plt stored for middle-late stage were significant higher than those of early stored stage (P＜0.05).@*CONCLUSION@#Morphology of EXO from LRA-Plt stored for middle-late stage was different from that stored for early stored stage. Moreover, the particle size distribution, contents of protein and RNA of EXO from LRA-Plt stored for middle-late stage were higher than those of early stored stage. A large amount of protein and RNA contained in EXO from LRA-Plt may participate in the multiple functions caused by platelet transfusion.

Distribution and antimicrobial resistance of Acinetobacter baumannii in a geriatric hospital from 2013 to 2016 / 中国感染控制杂志

Objective To analyze the clinical distribution and antimicrobial resistance change trend of Acinetobacter baumannii(AB) from a geriatric hospital in 2013-2016.Methods Specimen source,department distribution,and antimicrobial resistance of AB isolated from all patients in the hospital from 2013 to 2016 were analyzed retrospectively.Results From 2013 to 2016,1 712 strains of AB were isolated,AB isolation rates in 2013,2014,2015,and 2016 decreased year by year,which were 17.92％,17.17％,15.10％,and 11.81％,respectively.AB were mainly isolated from sputum (n =1 524,89.02％),followed by urine (n =79,4.61％) and blood (n=37,2.16％).The main departments of AB isolation were intensive care unit (n =798,46.61％),department of respiratory medicine (n =507,29.62％),and neurology department (n =156,9.11％).Resistance rates of AB to most antimicrobial agents increased in 2013-2016,resistance rates to compound sulfamethoxazole were low (25.68 ％-65.89 ％),followed by resistance rates to cefoperazone/sulbactam (54.74％-68.00％),resistance rates to imipenem were 71.40％-77.42％,to the other antimicrobial agents were all＞60％;in 2013-2016,resistance rates of AB to cefepime,cefoperazone/sulbactam,gentamicin,tobramycin,and compound sulfamethoxazole were significantly different (all P＜0.05).Conclusion Antimicrobial resistance rates of AB in this hospital is increasing,it is necessary to strengthen the monitoring,promote the rational use of antimicrobial agents,and block the infection and transmission of AB in hospital.

Research on displacement model of urine flow rate meter calibration device based on regression analysis / 医疗卫生装备

Objective To determine the relationship between the theory and actual displacement of the urine flow rate meter calibration device gear pump, and to construct a model between actual and theory displacement, in order to obtain actual displacement via theory displacement. Methods The weight of the standard water flow in each flow rate was measured by B2000S digital scales,and the scatter plot between the theory displacement and actual displacement was drawn to find the best fitted curve;then the MATLAB Curve Fitting Tool was used to obtain the best quasi relational equation. Results The error between the fitted displacement and the actual displacement of the gear bump was 1.494% in maximum and-0.010% in minimum, far less than 5% of the requirements for urinary flow meter measurement. Conclusion Based on regression analysis, the relationship between the theoretical displacement and actual displacement of the flow rate meter calibration device is established,the fitting error is small,and the method is simple and easy to achieve.

Significance of normal range urinary albumin to creatinine ratio in Chinese subjects with metabolic syndrome / 华中科技大学学报（医学）（英德文版）

This study was aimed to investigate clinical features of Chinese metabolic syndrome (MS) subjects with normal urinary albumin to creatinine ratio (UACR) and to estimate independent correlation factor for UACR. Data were drawn from a cross-sectional survey in participants having MS. The patients with different grade of albuminuria were divided into 4 groups according to the value of UACR (<10, 10-20, 21-30, >30 mg/g). All underwent biochemical tests. Bioelectrical impedance body fat content, islet β-cell function and insulin sensitivity were measured. Multivariable linear regression models were applied to further determine association between UACR and clinical factors with adjustment. Systolic blood pressure (SBP), diastolic blood pressure (DBP), mean arterial pressure (MAP), fasting plasma glucose (FPG), glycosylated hemoglobin (HbA1c), TG, fat mass, fat content and homeostasis model assessment for insulin resistance (HOMA-IR) were significantly higher in the group with UACR at 10-20 mg/g than those in the group with UACA lower than 10 mg/g (P<0.05). Multivariable linear regression showed that TG, HbA1c, waist-hip ratio (WHR) and SBP were independently associated with UACR. The patients with normal UACR had abnormal levels of MS components. The factors independently associated with UACR were TG, HbA1c, WHR and SBP.

Protective effect of Danxuetong injection against testicular injury after testis torsion/detorsion in rats / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the protective effect of Danxuetong injection (DXT, a combination of Danshen and Xueshuantong injections) against testicular ischemia-reperfusion injury following testis torsion/detorsion in rats.</p><p><b>METHODS</b>Thirty-two 4-week-old healthy male SD rats were randomly divided into four groups of equal number: sham operation, normal saline, single DXT injection, and successive DXT injection. The rat models of testicular ischemia-reperfusion injury were established by 2-hour 720-degree torsion/detorsion of the unilateral testis. At 6 weeks after modeling, the rats were killed and their testes were harvested for measure- ment of testicular coefficients, sperm counts, sperm motility, and the levels of total anti-oxidative capacity (T-AOC) , superoxide dismutase (SOD) , nitric oxide synthase (NOS) , and malondialdehyde ( MDA) in the testis tissue.</p><p><b>RESULTS</b>Compared with the rats of the normal saline group, those of the single DXT injection and successive DXT injection groups showed significant increases in the testicular coefficient (0.11 ± 0.03 vs 0.35 ± 0.04 and 0.40 ± 0.06, P < 0.05), sperm count ([0.46 ± 0.10] vs [1.44 ± 0.50] and [3.00 ± 1.28] x10(9)/ml, P < 0.05), sperm motility ([13.63 ± 14.04] vs [39.63 ± 5.04] and [76.31 ± 3.67]%, P < 0.05), the activity of SOD (72.76 ± 5.58 vs 116.25 ± 8.83 and 133.20 ± 13.84, P < 0.05), and the level of T-AOC (5.58 ± 1.07 vs 13.34 ± 5.81 and 19.21 ± 5.69, P < 0.05), but a remarkable decrease in the content of MDA (42.38 ± 8.94 vs 20.94 ± 5.65 and 15.02 ± 1.03, P < 0. 05) in the injured testes.</p><p><b>CONCLUSION</b>DXT can effectively rid the testis tissue of oxygen free radicals, improve sperm count and motility by antioxidation, and protect the testis tissue of prepubertal rats against testicular ischemia-reperfusion injury after testis torsion/detorsion. It also has a protective effect on the contralateral testis, and successive injection has a better effect than single injection of DXT.</p>

Formation of para-Bombay phenotype caused by homozygous or heterozygous mutation of FUT1 gene / 中国实验血液学杂志

This study was aimed to explore the molecular mechanisms for para-Bombay phenotype formation. The H antigen of these individuals were identified by serological techniques. The full coding region of alpha (1, 2) fucosyltransferase (FUT1) gene of these individuals was amplified by high-fidelity polymerase chain reaction (PCR). PCR product was identified by TOPO cloning sequencing. Analysis and comparison were used to explore the mechanisms of para-bombay phenotype formation in individuals. The results indicated that the full coding region of FUT1 DNA was successfully amplified by PCR and gel electrophoresis. DNA sequencing and analysis found that h1 (547-552delAG) existed in one chromosome and h4 (35C > T) existed in the other chromosome of NO.1 individual. Meantime, h1 (547-552delAG) was found in two chromosomes of NO.2 and NO.3 individual. It also means that FUT1 gene of NO.1 individual was h1h4 heterozygote, FUT1 gene of NO.2 and NO.3 individuals were h1h1 homozygote. It is concluded that homozygous and heterozygous mutation of FUT1 gene can lead to the formation of para-Bombay phenotype.

Effects of umbilical cord blood monocytes transplantation on EPO protein and oligodendrocyte progenitors in neonatal rats with hypoxic-ischemic brain damage / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the effects of umbilical cord blood monocytes (UCBMC) transplantation on erythropoietin (EPO) protein and oligodendrocyte progenitor cells in hypoxia-ischemia (HI) neonatal rats.</p><p><b>METHODS</b>Forty seven-day-old Sprague-Dawley rats were randomly divided into normal control (N), HI, UCBMC and HI+UCBMC groups (n=10 each). Hypoxic-ischemic brain damage (HIBD) model was prepared according to the Rice method. Twenty-four hours after hypoxia, the N and HI groups were injected with 2 μL phosphate buffered saline (PBS), and the UCBMC and HI+UCBMC groups were injected with 3×10(6) UCBMC via the lateral ventricle. EPO protein and oligodendrocyte progenitor cells in the subventricular zone of the injured brain were observed by EPO/DAPI and NG2/DAPI immunofluorescence double staining, and their correlation was analyzed.</p><p><b>RESULTS</b>Seven days after transplantation, there were more NG2(+)DAPI(+) and EPO(+)DAPI(+) cells in the HI+UCBMC group than in the UCBMC (P<0.05), N and HI groups (P<0.01). More NG2(+)DAPI(+) and EPO(+)DAPI(+) cells were observed in the UCBMC group compared with the N and HI groups (P<0.01). There were more NG2(+)DAPI(+) cells in the N group than in the HI group (P<0.01). The number of NG2(+)DAPI(+) cells was correlated with the number of EPO(+)DAPI(+) cells in the HI+UCBMC group (r=0.898, β=1.4604, P<0.01).</p><p><b>CONCLUSIONS</b>UCBMC can promote expression of oligodendrocyte progenitor cells, which is correlated with an increase in EPO protein and thus repairs brain white matter damage in neonatal rats with HIBD.</p>

Seroreactivity and immunogenicity of Tp0965, a hypothetical membrane protein of Treponema pallidum / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Treponema pallidum (T. pallidum) subsp. pallidum is the causative agent of syphilis. Analysis of recombinant antigens of T. pallidum led to the identification of potential candidate antigens for vaccine development and syphilis serodiagnosis. Tp0965 was predicted to be a membrane fusion protein and was found to be reactive with infected human sera in previous studies, but the results were controversial. In this research, the antigenicity and immunoreactivity of recombinant protein Tp0965 were assessed.</p><p><b>METHODS</b>T. pallidum subsp. pallidum (Nichols strain) was propagated and isolated and the genomic DNA was extracted. The Tp0965 gene was amplified by polymerase chain reaction (PCR). Then the recombinant protein Tp0965 was expressed in Escherichia coli and purified by nickel-nitrilotriacetic acid (Ni-NTA) purification system. The reactivities of protein Tp0965 were examined by immunoblot analysis and indirect enzyme-linked immunosorbent assay. The antisera against protein Tp0965 were obtained by immune rabbits and the immunogenicity of antisera were detected by indirect enzyme-linked immunosorbent assay.</p><p><b>RESULTS</b>Recombinant protein Tp0965 was expressed successfully in vitro. Immunoblot assay showed that the recombinant protein Tp0965 could be recognized by human syphilitic sera of all stages. Indirect enzyme-linked immunosorbent assay showed there were only 4 of 74 human syphilitic sera that failed to show reactivity to recombinant antigen Tp0965, and lack of reactivity of Tp0965 to all 28 uninfected sera. A low titer of antiserum against Tp0965 in immune rabbits could be detected after the third time of immunization.</p><p><b>CONCLUSIONS</b>The recombinant antigen Tp0965 shows excellent sensitivity for the reactivity with sera from syphilitic individuals at all stages. The results also demonstrate a potential application for the serodiagnosis of syphilis.</p>

Establishment of B lymphoblastoid cell lines of Miao pedigree with Bardet-Biedl syndrome / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To establish immortalized lymphoblastoid cell lines of a Miao core pedigree with Bardet-Biedl syndrome (BBS), in order to provide a long-term source of material for research.</p><p><b>METHODS</b>With Epstein-Barr virus transformation of B cells and addition of cyclosporine A to inhibit the activity of T cells, fresh anticoagulated blood samples with heparin were collected from 12 members of the core pedigree, and were used to establish the immortalized lymphoblastoid cell lines of B lymphocytes.</p><p><b>RESULTS</b>Twelve immortalized lymphoblastoid cell lines of the core BBS pedigree were obtained successfully.</p><p><b>CONCLUSION</b>The immortalized B lymphoblastoid cell lines of the Miao pedigree with BBS can preserve the whole genome information and provide long-term research materials for BBS study.</p>

Effects of Shengmai injection on testicular injury after torsion/detorsion in rats of different ages / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the different effects of Shengmai injection on testicular injury after testis torsion/detorsion in rats of different ages.</p><p><b>METHODS</b>Sixteen healthy male SD rats aged 3, 6 and 12 weeks were equally randomized into an experimental group (testicular torsion/detorsion plus Shengmai injection) and a control group (testicular torsion/detorsion plus saline). The rat models of testicular torsion were killed 24 h after surgery for the measurement of total antioxidant capacity (T-AOC), superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in the testis.</p><p><b>RESULTS</b>Compared with the controls, the 3- and 6-week-old rats of the experimental group showed no significant changes in T-AOC, SOD activity and MDA content (P > 0.05), while the 12-week-old experimental rats exhibited a remarkable increase in SOD and T-AOC and an obvious decrease in MDA content (P < 0.05).</p><p><b>CONCLUSION</b>Shengmai injection has a protective effect against acute ischemia-reperfusion testicular injury after torsion/detorsion in rats, but the effect varies with the age, more obvious in older ones.</p>

Nkx2-5 gene promotes expression of cardiac markers in 5-azacytidine-induced monolayer P19 cells / 第二军医大学学报

Objective: To study the role of Nkx2-5 gene in 5-azacytidine-induced differentiation of monolayer P19 cells into myocardial cells. Methods: The experiment was divided into two groups: an experimental group and a control group. The cells in the experimental group were P19 cells stably expressing Nkx2-5 gene, and cells in the control group were P19 cells. Under the monolayer-culture condition, the cells of two groups were induced by 5-azacytidine (1 μmol/L). The growth of cells were observed by inverted microscope. On the 4th day, 8th day, 12th day and 16th day after induction, RT-PCR was used to detect the expression of GATA-4, α-MHC and ANP gene. Results: In control group, there was no ANP expression after induction; GATA-4 expression was seen on the 8 th day, 12th day, and 16th day after induction; and α-MHC expression was found on the 12th day and 16 th day. In experimental group, the expression of GATA-4 was detected on the 4th day, 8th day, 12th day and 16 th day after induction; Alpha-MHC and ANP expression was noticed on the 8th day, 12th day and 16th day after induction. RT-PCR results showed that the expression of GATA-4 and α-MHC in the experimental group was earlier than that in the control group. And at all time points of observation, the expression of GATA-4 and α-MHC in the experimental group was significantly increased compared with that in the control group (P<0. 05,P<0. 01), except for α-MHC expression on the 12 th day. Conclusion: Nkx2-5 gene can promote 5-azacytidine-induced differentiation of monolayer P19 cells into myocardial cells.

Inhibitory effect of S-adenosylmethionine on the growth of human gastric cancer cells in vivo and in vitro / 癌症

<p><b>BACKGROUND AND OBJECTIVE</b>S-adenosylmethionine (SAM), the most important methyl donor in human body, is generally used to treat cholestasis in clinic. In recent years, SAM has been found to have inhibitory effects on breast cancer, liver cancer and colon carcinoma. This study was to investigate the inhibitory effects of SAM on human gastric cancer cells in vivo and in vitro, and the antitumor mechanisms.</p><p><b>METHODS</b>The effects of SAM on the proliferation of gastric cancer SGC-7901 and MKN-45 cells were determined by MTT assay. After SGC-7901 and MKN-45 cells were treated with 0, 2, and 4 mmol/L SAM for 72 h, the expression and methylation of c-myc and urokinase type plasminogen activator (uPA) were detected by reverse transcription-polymerase chain reaction (RT-PCR) and methylation-specific PCR (MSP). Tumor xenografts were established by injecting SGC-7901 cells subcutaneously in BALB/c nude mice. The mice were randomized into low concentration group [192 µmol/(kg · day)], high concentration group [768 µmol/(kg · day)], and control group [normal saline (NS)], and received peritoneal injection of relative reagents for 15 days. The tumor size was measured, the protein and mRNA expression of c-myc and uPA were detected by immunohistochemistry and RT-PCR, and the methylation of c-myc and uPA genes was detected by MSP.</p><p><b>RESULTS</b>SAM inhibited the growth of SGC-7901 and MKN-45 cells obviously and the effects were enhanced with the increase of SAM concentration and treatment time. The mRNA expression of c-myc and uPA in SGC-7901 cells and that of uPA in MKN-45 cells significantly decreased. The c-myc and uPA genes in SGC-7901 cells and uPA gene in MKN-45 cells were partly or completely methylated after SAM treatment. The tumor volume was significantly lower in low concentration group [(618.51 ± 149.27) mm³] and high concentration group [(444.32 ± 118.51) mm³] than in control group [(1018.22 ± 223.07) mm³] (both P < 0.01). The inhibitory rates of tumor growth were 39.26% in low concentration group and 56.36% in high concentration group. The protein and mRNA expressions of c-myc and uPA were remarkably reduced (all P < 0.01), and the hypomethylation of c-myc and uPA genes were reversed after SAM treatment.</p><p><b>CONCLUSIONS</b>SAM can inhibit the growth of human gastric cancer cells both in vivo and in vitro. The mechanism may be that SAM can reverse the hypomethylation of c-myc and uPA genes, reduce their expression, and then inhibit tumor growth.</p>

Exogenous Nkx2-5 gene expression induces the expression of cardiac markers during P19 cell differentiation in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the role of homeobox gene Nkx2-5 in cardiac myogenesis.</p><p><b>METHODS</b>Two P19 cell lines, namely cells transfected with exogenous expression of Nkx2-5 gene and non-transfected cells, were cultured in suspension for 4 days to induce cell aggregation, and the cell aggregates were transferred to the Petri dish for further adherent culture. On days 4, 8, 12 and 16 of adherent culture, the expressions of α-sarcomeric actin (α-SA) and cardiac troponin T (cTnT) protein were detected by immunocytochemistry, and the mRNA expressions of GATA-4, α-myosin heavy chain (α-MHC) and atrial natriuretic factor (ANF) genes by RT-PCR.</p><p><b>RESULTS</b>In the transfected cells, α-SA and cTnT protein expressions were detected on days 8, 12 and 16 of adhere culture, and their expressions increased gradually with time. α-SA and cTnT expression was significantly higher on day 16 than on day 8 of culture (P<0.01). RT-PCR analysis of the transfected cell showed the presence of GATA-4 expression on day 4 of adherent culture, and the expression increased on days 8 and 12 but decreased on day 16. ANF and α-MHC expressions were found on days 8, 12, and 16, increasing gradually over time and showing significant differences from those on day 4 (P<0.05 or P<0.01). The expression of α-MHC was significantly higher on days 12 and 16 than on day 8 (P<0.05 or P<0.01), and ANF expression was significantly higher on day 16 than on days 8 and 12 (P<0.01). The non-transfected cells were negative for the expressions of all these genes.</p><p><b>CONCLUSION</b>Exogenous expression of Nkx2-5 gene can induce P19 cells to express cardiac markers in vitro.</p>

Meta-analysis of the efficacy and safety of fluconazole in prophylaxis of fungal infection in very low birth weight infants / 中华儿科杂志

<p><b>OBJECTIVE</b>To evaluate the effect of fluconazole in prophylaxis of fungal infection in very low birth weight infants.</p><p><b>METHODS</b>PubMed, EMBASE, Ovid, China National Knowledge Infrastructure, Vip Chinese Periodical Database, Wanfang Chinese Periodical Database and Chinese Bio-medicine Database were searched for the case-control study on the effect of fluconazole in prophylaxis of fungal infection in very low birth weight infants from Jan. 1994 to Jan. 2009. Articles were evaluated according to inclusion criteria. Poor-quality studies were excluded, and RevMan 4.22 software was applied for investigating the heterogeneity among individual studies and calculating the pooled risk ratio (RR) and 95% confidence interval (CI).</p><p><b>RESULTS</b>Five eligible randomized clinical trials were included. Four studies were graded as "A" and one study was graded "B". Meta-analysis based on the included studies showed that the prophylactic fluconazole could significantly reduce the fungal colonization (RR: 0.32 and 95% CI: 0.23 to 0.44, P < 0.00001); and infections (RR: 0.44 and 95% CI: 0.29 to 0.65, P < 0.0001) in very low birth weight neonates. However, there was no statistically significant difference between the infants treated and not treated with prophylactic fluconazole in the neonatal mortality (RR: 0.68 and 95% CI: 0.43 to 1.07, P = 0.09) and the prophylactic use of fluconazole did not show any side-effects on the liver and bilirubin. None of the studies found any significant changes in the minimal inhibitory concentration of fluconazole in fungal isolates during the study period. There were different results about the emergence of resistance to fluconazole.</p><p><b>CONCLUSIONS</b>Meta-analysis of five randomized controlled trials suggest that prophylactic fluconazole reduces the incidence of fungal colonization and invasive fungal infection in very low birth weight infants. Further trials are needed to provide more precise evaluation on efficacy, and to assess the effect on mortality, neurodevelopment and the emergence of resistance to antifungal agents. Different NICU should have different policy on prophylactic fluconazole and also adjust the policy at different time according to the incidence of fungal infection and antifungal drug resistance.</p>

Expression of Cx43 and Pax3 in the small intestinal muscular layers of early human embryos / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the patterns of Cx43 and Pax3 protein expressions in the small intestinal muscular layers of human embryo during early development.</p><p><b>METHODS</b>Immunohistochemistry with SABC method was employed to examine the expression of Cx43 and Pax3 proteins in the muscular layers of the small intestine in early human embryos in the second to fourth months of gestation.</p><p><b>RESULTS</b>In the second month of gestation, the muscle layer of the small intestine was negative for Cx43 and Pax3 protein expressions. In the third month, Cx43 and Pax3 expressions were negative in the inner circular muscle layer, but some positive cells were found in the longitudinal muscle layer and the myenteric plexus. In the fourth month, positive expression of Cx43 and Pax3 proteins were seen in the entire muscle layer.</p><p><b>CONCLUSION</b>Cx43 and Pax3 proteins are closely related to the growth and development of the cells and tissues in the small intestinal muscle layer in human embryos.</p>

Changes and clinical significance of Toll-like receptor 2 and 4 expression in neonatal infections / 中华儿科杂志

<p><b>OBJECTIVES</b>Neonates are vulnerable to various infections because of their immature immune responses. Toll-like receptors could induce immune responses, both the innate and the acquired immune responses. The aim of the present study was to investigate the changes of TLR2 and TLR4 in neonatal infections, and to determine their roles in anti-infection immune reaction.</p><p><b>METHODS</b>A total of 200 infants were divided into six groups: sepsis group (n = 21), bacterial pneumonia group (n = 70), bacterial meningitis group (n = 17), urinary tract infection group (n = 38), congenital syphilis group (n = 11) and non-infection group (n = 48). The TLR mRNA was determined by RT-PCR. The protein expression of TLR and the percentage of TLR positive cells were evaluated through flow cytometric analysis.</p><p><b>RESULTS</b>1. The TLR2 mRNA expression increased significantly in the sepsis group (6.14 +/- 0.80), most significantly in the Gram positive sepsis group (6.43 +/- 0.74). TLR2 mRNA expression was also significantly higher in the bacterial pneumonia group (5.49 +/- 0.62), the bacterial meningitis group (5.61 +/- 0.60) and the congenital syphilis group (5.89 +/- 0.38). TLR2 protein expression was the highest in the sepsis group and significantly increased in the bacterial pneumonia group, bacterial meningitis group and the congenital syphilis groups as well, all were higher than the TLR2 protein expression of the non-infectious group (1.27 +/- 0.75). The TLR2 protein expression in the Gram positive bacterial sepsis group was 2.54 +/- 0.68, that of Gram negative bacterial sepsis group was 1.25 +/- 0.51 (P < 0.05). The percentage of TLR2 positive cells in the neonatal infection group was (70.95 +/- 20.15)%, which did not differ significantly from that of non-infection group. 2. The mRNA expression of TLR4 was the highest in the sepsis group (6.20 +/- 1.59), while that in the Gram negative bacterial sepsis group was 6.78 +/- 1.79, higher than that of the Gram positive bacterial sepsis group, 5.39 +/- 0.78, (t = 2.29, P = 0.037). TLR4 mRNA expression increased significantly in the bacterial pneumonia group (5.33 +/- 1.07), the bacterial meningitis group (5.87 +/- 0.70) and the urinary tract infection group (5.38 +/- 0.91). There were no significant differences in TLR4 protein expression among these groups. The percentage of TLR4 positive cells in the neonatal infection groups was (0.71 +/- 0.31)%, higher than that of non- infection group (0.29 +/- 0.36)%. 3. In the Gram positive bacterial sepsis group, the mRNA expression of TLR2 (6.43 +/- 0.74) was higher than the mRNA expression of TLR4 (5.39 +/- 0.78), (t = 1.56, P = 0.024). In the Gram negative bacterial sepsis group, the mRNA expression of TLR4 (6.78 +/- 0.79) was significantly higher than the mRNA expression of TLR2 (5.64 +/- 0.68) (t = 2.63, P = 0.011). In the sepsis group, the TLR2 protein expression was significantly higher than the expression of TLR4 (t = 1.06, P = 0.044). The percentage of TLR4 positive cells was lower than the percentage of TLR2 positive cells among all these groups, P < 0.01. 4. Correlation analysis on gestational age and the mRNA expression, the protein expression and the percentage of TLR2 and TLR4 positive cells among all these groups did not show any statistical significance.</p><p><b>CONCLUSIONS</b>The mRNA and the protein expression of TLR2 and the mRNA expression of TLR2 increased significantly in the studied neonatal infection groups, especially in the severe sepsis groups. The mRNA expression of TLR2 increased mainly in the Gram positive bacterial infection groups, and the mRNA expression of TLR4 increased in the Gram negative bacterial infection groups, suggesting that both the TLR2 and TLR4 signal pathway took part in the immune mechanism of neonatal infection, providing new idea and experimental basis for further understanding of immune mechanism of neonatal infection.</p>

Comparative proteome analysis of human papillomavirus-infected cervical specimens and the difference between the high- and low-risk genotypes of human papillomavirus / 中国医学科学院学报

<p><b>OBJECTIVE</b>To perform an comparative proteome analysis of human papillomavirus-infected cervical specimens and to investigate different expressions between high- and low-risk genotypes.</p><p><b>METHODS</b>The cervical specimens were divided into two groups (cervical intraepithelial neoplasia group and condyloma acuminatum group) according to their genotypes. Using comparative proteome technology, high-risk human papillomavirus-infected cervical intraepithelial neoplasia, low-risk human papillomavirus-infected condyloma acuminatum, and normal cervical intraepithelial tissue were compared. The differential expression protein spots were identified by mass spectrometry.</p><p><b>RESULTS</b>Totally 26 differential spots were selected and analyzed, and 22 peptide mass fingerprints (PMF) maps were obtained by MALDI-TOF-MS. Eighteen proteins were preliminarily identified after searching the NCBInr database. The function information of these 18 proteins mainly involved cell metabolism, signal transduction, cell secretion, cell cytoskeleton construction, cell proliferation, and apoptosis.</p><p><b>CONCLUSION</b>The proteomic expressions after the cervical infection of high- or low-risk genotype of human papillomavirus are obviously different.</p>

Retrospective Analysis of Clinic and Epidemiology of Respiratory Syncytial Virus Pneumonia in 309 Neonates / 实用儿科临床杂志

Objective To explore the characteristics of clinic and epidemiology of neonatal respiratory syncytial virus(RSV) pneumonia in newborn infants.Methods Three hundred and nine neonates(male 196,female 113) who were admitted to children's hospital of Fudan University and identified as having RSV pneumonia from Jan.2004 to Dec.2008 were enrolled.RSV antigen was detected in exfoliated respiratory cells by direct immunofluorescence.All the clinical data were collected and subjects and they were divided into different groups based on path of infection,gestational age and RSV status.Results From case notes over the 5 years,309 eligible neonates with RSV infection were identified.Male term neonates were more likely to acquire infection,as compared to female,1.71.0.The incidence of RSV infection peaked in winter and spring.Compared with community acquired infection group,hospital acquired infection group had more premature infants(23.2% vs 8.7%,P=0.002),lower birth weight[(3 010.8?852.8) g vs(3 153.2?943.4) g,P=0.026],much longer mean length of hospital stay[(19.0?8.0) d vs(12.2?4.5) d,P=0],more expensive cost[(5 646.4?3632.2) RMB vs(4 175.8?2 879.2) RMB,P=0] and later occurrence day[(21.3?8.6) d vs(15.8?6.0) d,P=0].Compared with simple RSV infection group,mixed infection group had more expensive cost[(6 063.1?3 085.4) RMB vs(4 513.2?3 860.8) RMB,P=0.047] and more oxygen use(40.0% vs 25.7%,P=0.006).The clinical characteristics of patients with RSV pneumonia varied.Preterm group compared with term group more frequently exhibited apnea and cyanosis(20.0% vs 0.7%,42.9% vs 22.7%),respectively.Forty-two percent of them had bacterial infection,while 13.3% of them had infection of other part of body.The prognosis of most RSV(99.0%) infections was good.Conclusions RSV was an important cause of respiratory tract infections during the last five years.Clinical characteristics of RSV infections were atypical.Hospital acquired RSV infection was more severe than the community acquired infection in neonates.There are no specific,effective interventions for treating RSV infections,so preventive measures are most important.