[Application value of serum protein indicators in constructing the early prediction model for the prognosis of patients with pulmonary tuberculosis].

Objective: To analyze the clinical significance of laboratory examination indicators as the key prognostic factors and to construct an early prediction model for prognosis assessment of pulmonary tuberculosis patients. Methods: The basic information, biochemical indexes and blood routine items of 163 tuberculosis patients (144 males and 19 females, aged 41-70 years, with an average age of 56 years) and 118 healthy persons who underwent physical examination (101 males and 17 females, aged 46-64 years, with an average age of 54 years) in Suzhou Fifth People's Hospital from January 2012 to December 2020 were retrospectively collected. According to the presence of Mycobacterium tuberculosis after six months of treatment, the enrolled patients were divided into a cured group (96 cases) and a treatment failure group (67 cases). To analyze the baseline levels of laboratory examination indicators between these two groups, we screened the key predictors and the binary logistic regression method in SPSS statistics software was used to construct the prediction model. Results: The baseline levels of total protein, albumin, prealbumin, glutamic-pyruvic transaminase, erythrocyte, hemoglobin and lymphocyte were significantly higher in the cured group than in the treatment failure group. After 6 months of treatment, the indexes of total protein, albumin and prealbumin increased significantly in the cured group, but remained at the low levels in the treatment failure group. Receiver operating characteristic (ROC) curve analysis showed that total protein, albumin and prealbumin as independent predictors for forecasting the prognosis of pulmonary tuberculosis patients had the highest prediction accuracy. Logistic regression analysis showed that the combination of these three key predictors could construct the best early prediction model for assessing the prognosis of pulmonary tuberculosis patients, with a prediction accuracy of 0.924 (0.886-0.961), sensitivity of 75.0%, specificity of 94%, showing an ideal prediction accuracy. Conclusions: The routine test indexes of total protein, albumin and prealbumin show good application value in the construction of early prediction model for prognosis evaluation of pulmonary tuberculosis treatment. The combined prediction model consisting of total protein, albumin and prealbumin is expected to provide a theoretical basis and reference model for precision treatment and prognosis assessment of tuberculosis patients.

[Pre-operative prognostic nutritional index as a predictive factor for prognosis in non-metastatic renal cell carcinoma treated with surgery].

OBJECTIVE: To evaluate the implications of the prognostic nutrition index (PNI) in non-metastatic renal cell carcinoma (RCC) patients treated with surgery and to compare it with other hematological biomarkers, including neutrophil to lymphocyte ratio (NLR), platelet to lymphocyte ratio (PLR), and systemic immune inflammation index (SII). METHODS: A cohort of 328 non-metastatic RCC patients who received surgical treatment between 2010 and 2012 at Peking University First Hospital was analyzed retrospectively. Receiver operating characteristic (ROC) curve analysis was used to determine the optimal cutoff values of the hematological biomarkers. The Youden index was maximum for PNI was value of 47.3. So we divided the patients into two groups (PNI≤ 47. 3 and >47. 3) for further analysis. Categorical variables [age, gender, body mass index (BMI), surgery type, histological subtype, necrosis, pathological T stage and tumor grade] were compared using the Chi-square test and Student' s t test. The association of the biomarkers with overall survival (OS) and disease-free survival (DFS) was analyzed using Kaplan-Meier methods with log-rank test, followed by multivariate Cox proportional hazards model. RESULTS: According to the maximum Youden index of ROC curve, the best cut-off value of PNI is 47. 3. Low level of PNI was significantly associated with older age, lower BMI and higher tumor pathological T stage (P < 0.05). Kaplan-Meier univariate analysis showed that lower PNI was significantly correlated with poor OS and DFS (P < 0.05). In addition, older age, lower BMI, tumor necrosis, higher tumor pathological T stage and Fuhrman grade were significantly correlated with poor OS (P < 0.05). Cox multivariate analysis showed that among the four hematological indexes, only PNI was an independent factor significantly associated with OS, whether as a continuous variable (HR=0.9, 95%CI=0.828-0.978, P=0.013) or a classified variable (HR=2.397, 95%CI=1.061-5.418, P=0.036). CONCLUSION: Low PNI was a significant predictor for advanced pathological T stage, decreased OS, or DFS in non-metastatic RCC patients treated with surgery. In addition, PNI was superior to the other hematological biomar-kers as a useful tool for predicting prognosis of RCC in our study. It should be externally validated in future research before the PNI can be used widely as a predictor of RCC patients undergoing nephrectomy.

[Clinical effects of pre-expanded anterior perforator flap of transverse cervical artery and pre-expanded thoracic random flap in reconstructing extensive facial and cervical scar in relay].

Objective: To explore the clinical effects of pre-expanded anterior perforator flap of transverse cervical artery in extensive facial and cervical scar reconstruction and contralateral pre-expanded thoracic random flap in relay in donor site repair. Methods: A retrospective cohort study was conducted. From May 2008 to December 2018, 10 patients with extensive facial and cervical scar after burns were treated in the Fourth Medical Center of PLA General Hospital, including 8 males and 2 females, aged 10-55 years. In the first stage of operation, two skin and soft tissue expanders of the same volume (with rated capacity of 250-600 mL) were respectively placed in the right side and left side of the chest according to the size of scar, and then the skin was expanded. The total amount of normal saline injected was 2 to 4 times of the rated capacity of the expander. In the second stage, the defect with area of 12 cm×8 cm-23 cm×15 cm caused by scar resection and release was repaired with unilateral pre-expanded anterior perforator flap of transverse cervical artery with area of 12 cm×9 cm-24 cm×16 cm. The contralateral pre-expanded thoracic random flap with the same area as that of the above-mentioned perforator flap was extended to repair the secondary defect with area of 8 cm×6 cm-17 cm×14 cm formed after transfer of the above-mentioned perforator flap. The exploration of perforating branch of transverse cervical artery, flap transfer and survival, injury repair, and complications were observed. The appearance and related function of donor and recipient sites and satisfaction of patients were followed up. Results: The perforating branches of transverse cervical artery appeared stably in the 10 patients. All the flaps were transferred to the recipient area without tension and survived. Both facial and cervical injuries were repaired successfully with no common complications. During the follow-up of 6 months-8 years, the color and texture of the pre-expanded anterior perforator flap of transverse cervical artery matched with the surrounding tissue, the functions of head raising and neck rotation of patients were significantly improved compared with those before operation, the color and texture of the flap transplanted in the first donor site matched with the original skin, linear scar left at the surgical incision, and 9 patients were satisfied with the restoration of the appearance and function of donor and recipient sites. Conclusions: The color and texture of the pre-expanded anterior perforator flap of transverse cervical artery match well with the face and neck, and the repairable area is large. After the perforator flap is removed, the secondary wound can be repaired with the pre-expanded thoracic random flap at the same time, and the injury of the chest donor site is alleviated. This relay repair method is a good choice for reconstructing extensive facial and cervical scar.

Conjugating MMAE to a novel anti-HER2 antibody for selective targeted delivery.

OBJECTIVE: To investigate the target delivery properties of RC48-ADC, a novel antibody drug conjugate (ADC) comprising cytotoxic monomethyl auristatin E (MMAE) and an anti-human epidermal growth factor receptor 2 (HER2) antibody tethered via valine-citrulline linker, in vitro and in vivo. MATERIALS AND METHODS: Dissociation rate of MMAE from RC48-ADC was used as an estimate of its stability in serum. Cytotoxicity of the antibody and RC48-ADC towards multiple cell lines was measured. Subcellular distribution of the drug was determined by fluorescence imaging. The mechanism of lysosome targeting was verified. Endocytic pathways of RC48-ADC were assessed by the cellular fluorescence intensity of fluorescently-labelled drugs. Intracellular and extracellular distribution of MMAE was analysed after RC48-ADC or MMAE administration to characterize MMAE release. The serum and tumour concentration of MMAE was compared after tail-vein injection of RC48-ADC into tumour-bearing mice. RESULTS: RC48-ADC was highly stable in human serum. HER2-overexpressed cell line SK-BR-3 proliferation was stronger when suppressed by RC48-ADC than by the naked antibody. Both RC48-ADC and naked antibody were internalized via caveolae-mediated and clathrin-mediated endocytosis and concentrated in lysosomes. Higher HER2 expression was associated with enhanced uptake and intracellular release of conjugated MMAE; free MMAE could kill tumour cells via the bystander effect. Although serum RC48-ADC concentration was higher than that in tumours, exposure of MMAE in tumours was ~200 times higher than in serum, which rationalized the reduced toxicity of RC48-ADC. CONCLUSIONS: In vitro and in vivo experiments confirmed the targeted transport and release of RC48-ADC; it could selectively deliver MMAE to the targeted HER2-positive cell or tumour tissue, which could reduce off-target toxicity and enhance anti-tumour potency in humans.

Impulse Control and ERP Characteristics of Patients with Mental Disorder Caused by Traumatic Brain Injury.

ABSTRACT: Objective To explore the impulse control and event-related potential （ERP） characteristics of patients with mental disorders caused by traumatic brain injury （TBI） in forensic psychiatry identification and to provide objective auxiliary indicators for forensic psychiatry identification. Methods Thirty patients （TBI group） with mental disorders caused by traumatic brain injury, who were identified as mild psychiatric impairment by judicial psychiatry, including 24 males and 6 females, as well as the thirty people in the control group participated in the study. All the participants completed Barratt Impulsiveness Scale-11 （BIS-11） and ERP induced by Go/NoGo tasks. BIS-11 and ERP data were collected and analyzed. Results The results of the BIS-11 showed that the total score and subscale scores of the TBI group were higher compared to the control group （P<0.05）. Moreover, the TBI group exhibited significantly lower NoGo-N2 amplitude and lower NoGo-P3 amplitude than the control group. The NoGo-N2 amplitude was larger than the Go-N2 amplitude, and the NoGo-P3 amplitude was larger than the Go-P3 amplitude in both groups （P<0.05）. Conclusion Traumatic brain injury could impair impulse control of mild psychiatric impairment patients, and the amplitudes of NoGo-N2 and NoGo-P3 could be important parameters to evaluate the impulse control of patients with mental disorders caused by traumatic brain injury.

[Comparison of safety and effectiveness between retroperitoneal laparoscopic tumor aspiration and laparoscopic partial nephrectomy in the treatment of renal angiomyolipoma].

OBJECTIVE: To compare the safety and treatment effectiveness of retroperitoneal laparoscopic tumor aspiration and laparoscopic partial nephrectomy (LPN) in the treatment of renal angiomyolipoma (RAML). METHODS: We retrospectively reviewed the clinical data of patients with pathologically confirmed RAML who received operation between August 2010 and August 2016 in the Department of Urology, Peking University First Hospital. Among them, a series of 121 patients were included in this trial according to the inclusion criteria, of which 74 cases could be collected and followed-up effectively. Based on the detailed surgical route, the 74 patients were divided into groups A and B: group A, which underwent retroperitoneal laparoscopic tumor aspiration, included 43 cases; group B, which received retroperitoneal LPN, included 31 cases. Patient demographics, intraoperative variables and postoperative outcomes were reported and compared between the groups. RESULTS: No statistical difference was detected in both groups before the treatment. Intraoperatively, the mean estimated blood loss was 48.7 mL in group A and 102.9 mL in group B, and the mean operative time was 70.1 min (21.2 min of warm ischemia time included) in group A and 103.6 min (28.5 min of warm ischemia time included) in group B, which were both statistically different. In group A, no complications occurred and yet 2 complications of transfusion and 1 complication of urine leakage were discovered in group B, although all finally recovered only with conservative treatment. A statistical difference was observed in the complication rates. Post-operatively, the mean serum creatinine level was 1.13 mg/dL in group A, and the level was 1.08 mg/dL in group B, in which no evident difference was detected. In a mean 52.6-months' follow-up, a recurrence of 3 cases in group A (7.0%) and a recurrence of 2 cases in group B (6.5%) were reported. No evident difference was also detected between the groups in the tumor recurrence rates. CONCLUSION: Due to the improvements in the intraoperative blood loss and operative time, retroperitoneal laparoscopic tumor aspiration may be provided with more potential advantages in the safety, also with equal efficacy of lower tumor recurrence rates when compared with the traditional retroperitoneal LPN in the treatment of RAML.

[A comparison of the outcome of therapy between fractional flow reserve guided- versus coronary angiography guided-strategy in acute coronary syndrome patients with moderate coronary lesions].

OBJECTIVE: To compare the outcomes between coronary angiography (CAG ) guided- and fractional flow reserve (FFR) guided-strategy in acute coronary syndrome (ACS) patients with moderate lesions. METHODS: Totally, 249 ACS subjects with moderate lesions examined by CAG in Beijing Anzhen Hospital from July 1, 2014 to July 30, 2015 were included in the present analysis. Among them, 98 patients were further examined by FFR and 151 were not. All the patients were treated with medication either guided by CAG or by FFR. Subjects were followed up for an average of 10 months. The end points included death, nonfatal myocardial infarction (MI), target vessel revascularization (TVR), and hospitalization costs. Major adverse cardiac events (MACE) were defined as death, nonfatal MI, and TVR. RESULTS: At the end of follow-up, 29 patients had MACE with 6 cases (6.5%) in the FFR-guided group, and 23 cases (16.2%) in the CAG-guided group (P=0.036). Patients treated with FFR-guided strategy had significantly lower rate of TVR than those treated with CAG-guided strategy (5.4% vs 14.8%, P=0.045). No statistical difference was observed in nonfatal MI (2.2% vs 3.5%, P=0.242) between the two groups, and no cardiac death occurred in the two groups. However, the rate of patients treated with stains (P=0.033) and the hospitalization costs (P=0.001) were significantly increased in the FFR-guided group. CONCLUSIONS: FFR-guided strategy for patients with ACS results in lower TVR and MACE, but higher cost when compared with CAG-guided strategy.

Harnessing the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated Cas9 system to disrupt the hepatitis B virus.

The current therapies to treat hepatitis B virus (HBV) infection are limited. Recently, clustered regularly interspaced short palindromic repeat (CRISPR) systems, originally identified in bacteria and archaea, have been found to consist of an RNA-based adaptive immune system that degrades complimentary sequences of invading plasmids and viruses. Here, we studied the effects of the CRISPR/CRISPR-associated Cas9 system that was targeted to the surface antigen (HBsAg)-encoding region of HBV, both in a cell culture system and in vivo. The HBsAg levels in the media of the cells and in the sera of mice were analyzed by a quantitative enzyme-linked immunosorbent assay. The HBV DNA levels were assessed by quantitative PCR and HBsAg expression in mouse livers was assessed by an immunohistochemical assay. The amount of HBsAg secreted in the cell culture and mouse serum was reduced by CRISPR/Cas9 treatment. Immunohistochemistry analyses showed almost no HBsAg-positive cells in the liver tissue of CRISPR/Cas9-S1+X3-treated mice. The CRISPR/Cas9 system efficiently produced mutations in HBV DNA. Thus, CRISPR/Cas9 inhibits HBV replication and expression in vitro and in vivo and may constitute a new therapeutic strategy for HBV infection.

Genome-wide survey and phylogenetic analysis on subunit sequences of eukaryotic DNA polymerase delta.

DNA polymerase d is not only the major replicative enzyme in eukaryotic chromosomal DNA synthesis but is also the primary polymerase for most DNA repair pathways. However, the subunit composition of polymerase d varies in different organisms. While polymerase d in many eukaryotic species has all 4 subunits (POLD1, 2, 3, and 4), many other organisms do not possess POLD4. Whether POLD4 is indispensable and why these differences exist are unknown. In the present study, we identified the POLD4 protein sequences of 218 eukaryotic species and determined the POLD1, 2, and 3 protein sequences of 55 species representing various taxonomic groups. No insect and nematode species examined possessed POLD4. Approximately 80% of protozoan species did not contain POLD4. Nearly 50% of fungal species did not contain POLD4. Other animal and plant species are expected to contain POLD4. Phylogenetic analyses of POLD1, 2, 3, and 4 sequences revealed that most animal and plant species inherited DNA polymerase d from protozoa, whereas some other animal and plant species may have inherited polymerase d directly from fungi. Because a large number of protozoan and fungal species do not possess POLD4, current insect and nematode species lacking POLD4 may have evolved from ancestor protozoan species lacking POLD4; thus, other protozoan and animal species lacking POLD4 may share a similar evolutionary history. Future studies should examine the origin and indispensability of POLD4 in various organisms.

Inelastic x-ray scattering study of exciton properties in an organic molecular crystal.

Excitons in a complex organic molecular crystal were studied by inelastic x-ray scattering (IXS) for the first time. The dynamic dielectric response function is measured over a large momentum transfer region, from which an exciton dispersion of 130 meV is observed. Semiempirical quantum chemical calculations reproduce well the momentum dependence of the measured dynamic dielectric responses, and thus unambiguously indicate that the lowest Frenkel exciton is confined within a fraction of the complex molecule. Our results demonstrate that IXS is a powerful tool for studying excitons in complex organic molecular systems. Besides the energy position, the IXS spectra provide a stringent test on the validity of the theoretically calculated exciton wave functions.

Studies of micro-, kappa-, and delta-opioid receptor density and G protein activation in the cortex and thalamus of monkeys.

The aim of this study was to investigate the relative density of micro -, kappa-, and delta-opioid receptors (MOR, KOR, and DOR) and guanosine 5'-O-(3-[35S]thio)triphosphate ([35S]GTPgammaS) binding stimulated by full agonists in cortical and thalamic membranes of monkeys. The binding parameters [Bmax (femtomoles per milligram)/Kd (nanomolar)] were as follows: [3H][d-Ala2,N-Me-Phe4,Gly5-ol]-enkephalin (DAMGO) (MOR; 80/0.7), [3H]U69593 [(5alpha,7alpha,8beta)-(-)-N-methyl-N-(7-(1-pyrrolidinyl)-1-oxaspiro(4,5)dec-8-yl) benzeneacetamide] (KOR; 116/1.3), and [3H][d-Pen2,d-Pen5]-enkephalin (DPDPE) (DOR; 87/1.3) in the cortex; [3H]DAMGO (147/0.9), [3H]U69593 (75/2.5), and [3H]DPDPE (22/2.0) in the thalamus. The relative proportions of MOR, KOR, and DOR in the cortex were 28, 41, and 31% and in the thalamus were 60, 31, and 9%. Full selective opioid agonists, DAMGO (EC50 = 532-565 nM) and U69593 (EC50 = 80-109 nM) stimulated [35S]GTPgammaS binding in membranes of cortex and thalamus, whereas SNC80 [(+)-4-[(alphaR)-alpha-((2S,5R)-4-allyl-2,5-dimethyl-1-piperazinyl)-3-methoxybenzyl]-N,N-diethyl-benzamide] (DOR; EC50 = 68 nM) was only active in cortical membranes. The magnitudes of [35S]GTPgammaS binding stimulated by these agonists were similar in the cortex, ranging from 17 to 25% over basal binding. In the thalamus, DAMGO and U69593 increased [35S]GTPgammaS binding by 44 and 23% over basal, respectively. Opioid agonist-stimulated [35S]GTPgammaS binding was blocked selectively by antagonists for MOR, KOR, and DOR. The amount of G protein activated by agonists was highly proportional to the relative receptor densities in both regions. These results distinguish the ability of opioid agonists to activate G proteins and provide a functional correlate of ligand-binding experiments in the monkey brain. In particular, the relative densities of opioid receptor binding sites in the two brain areas reflect their functional roles in the pharmacological actions of opioids in the central nervous system of primates.

["Target secondary structural motif" in the action of antisense oligodeoxynucleotides].

AIM: To optimize the antisense drug design based on the methods of secondary structure prediction of target mRNA by computer and the quantitative structure-activity relationship analysis. METHODS: The secondary structures of mRNA were predicted by the software RNAstructure, then the antisense phosphorothioate oligodeoxynucleotides (AS-ODN) were designed against the secondary structural elements. The in vitro anti-tumor bioactivity of AS-ODN was evaluated by A549 lung carcinoma cell line. The multiple regression was performed with the computer program SPSS. RESULTS: AS-ODN with high bioactivity concentrated on some local secondary structural motifs that were composed of several secondary structural elements, designated here as "target secondary structural motif" (target motif). The target motifs were relatively stable in the whole mRNA structures, but there were one or more unstable secondary structural elements (free energy > 0) such as internal loops, knots, and hairpins, especially the bulge loops in the motif. Bioactivities of AS-ODN targeting different "target motifs" were statistically different (P < 0.01) while AS-ODN against the same "target motif" were with similar effects. CONCLUSION: The concept of the "target motif" was helpful for optimizing the antisense drug design and might be useful for discovering the local function of mRNA and designing oligonucleotide probes and primers.

Application of secondary structure prediction in antisense drug design targeting protein kinase C-alpha mRNA and QSAR analysis.

AIM: To optimize the design of antisense drug targeting protein kinase C-alpha (PKC-alpha) mRNA and obtain better antisense drugs than ISIS3521 that is undergoing clinical trials. METHODS: RNAstructure (version 3.21, 1999) was utilized to predict the optimal and suboptimal secondary structures of human PKC alpha mRNA (GenBank, X52479), and 29 antisense phosphorothioate oligodeoxynucleotides (S-ODN) targeting the secondary structural elements, 3 partly matched S-ODN and 1 scrambled 3521 were designed. ISIS3521 was set as positive control. Mean (n = 3-5) 50% inhibitory effects on proliferation of A549 cells (IC50) of S-ODN were evaluated. Free energies (delta G degree 37) relating to the target secondary structural elements were calculated according to the nearest neighbor model. The quantitative structure-activity relationship (QSAR) analysis through multiple regression was obtained by SPSS. RESULTS: Three S-ODN; (5'-AGCCCA-GCCGCTTGGCTGGG-3', 5'-AGGAGTGCAGCTGC-GTCAAG-3', 5'-TCAGAGGG-ACTGATGACTTT-3') had lower IC50[(48 +/- 7), (50 +/- 4), (64 +/- 2.7) nmol.L-1, respectively] than that of ISIS3521 [(81 +/- 25) nmol.L-1]. The number of bases comprising the target secondary structural element bulge loop, internal loop, and knot, the free energy of S-ODN (delta G degree 37S), and reaction (delta G degree 37R) were important parameters in QSAR equation. In the multiple regression, R was 0.68, P = 0.0193. Not tally with the equation, two S-ODN (5'-TCAAATGGAGG-CTGCCCGGC-3', 5'-AAAACGTCAGCCATGGTCCC-3') with favorable target structures and delta G degree 37 did not behave good activities. CONCLUSION: Computer aided design was helpful to obtain S-ODN with better in vitro effect than current positive drug. The degree of instability of secondary structural elements and delta G degree 37 were important factors for drug activity. Other important factors needed for further investigation.

Pharmacokinetics and peripheral platelet counts after a single dose of recombinant human thrombopoietin in rhesus monkeys.

AIM: To study the pharmacokinetics and the change of peripheral platelet counts after a single dose of recombinant human thrombopoietin (rhTpo). METHODS: After i.v. or s.c. injections of rhTpo in 12 rhesus monkeys, rhTpo concentration in serum was determined by ELISA. Platelets were counted by automatic microcell counter. RESULTS: The terminal half-lives of rhTpo were 12-18 h. AUC following s.c. were linearly increased with dose, while Cls were 0.061, 0.08, and 0.07 L.kg-1.h-1 in s.c. 0.5, 2, and 8 micrograms.kg-1 groups, respectively. Bioavailability was 0.50 +/- 0.18 after s.c. Single dose of rhTpo was associated with an increase in platelets (55.9%-107.4%, P < 0.05) in a dose-related manner. The peak response and the sustained days of platelet increase were dose-related. The degree of platelet increase (% x time) correlated with the systemic exposure to rhTpo (C x time). CONCLUSION: rhTpo behaved as a linear pharmacokinetics in the monkey within dose range of 0.5-8 micrograms.kg-1.

A novel factor produced by placental cells with activity against HIV-1.

The factors controlling the dynamics of HIV-1 transmission from mother to infant are not clearly known. Previous studies have suggested the existence of maternal and placental protective mechanisms that inhibit viral replication in utero. Preliminary studies from our laboratory revealed that supernatant from placental stromal cells protected HIV-1-infected PBMC from virus-induced apoptosis and suppressed virus production. We have attempted to characterize the antiviral activity of this placental factor (PF) and delineate the stages of HIV-1 replication affected. This activity was not due to the presence of any known cytokine reported to have anti-HIV effect. Direct exposure to PF had no suppressive effect on the infectivity of cell-free HIV-1, and envelope-mediated membrane fusion appeared to be unaffected. Western blot analysis of HIV-1 from infected PBMC treated with PF revealed that expression of all viral proteins was reduced proportionately, both intracellularly and in released virions. However, exposure of HIV-1-infected cells to PF resulted in production of virions with 10-100-fold-reduced infectivity. PF-treated virions contained two- to threefold reduced ratios of cyclophilin A:Gag protein as compared with untreated virus. Reduced cyclophilin A content resulting in decreased binding of cyclophilin A to Gag could account, in part, for the observed reduction in infectivity. Our results suggest that placental cells produce an antiviral factor that protects the fetus during gestation and may have therapeutic potential.

Chemokine-independent in vitro resistance to human immunodeficiency virus (HIV-1) correlating with low viremia in long-term and recently infected HIV-1-positive persons.

Chemokines have been implicated as protective factors against human immunodeficiency virus (HIV) infection, competing for binding to receptors that also function as coreceptors for HIV. In this study of HIV-positive donors, peripheral blood mononuclear cell (PBMC) culture resistance to endogenous and exogenous HIV correlated with low plasma viremia and high in vitro RANTES production. However, resistant cells were not rendered susceptible by neutralization of C-C chemokines, and addition of C-C chemokines did not consistently suppress endogenous virus or exogenous HIV-1MN. In contrast, CD8 T cell depletion markedly decreased the frequency of resistant cultures without reducing C-C chemokine production. Among newly infected persons, half exhibited phenotype switching from preinfection susceptibility to postinfection resistance, suggesting that genetically predetermined constitutive cytokine production or allelic receptor expression are not generally responsible for in vitro resistance and nonprogression.

Diagnosis of human immunodeficiency virus infection using citrated whole blood.

Standard isolation of human immunodeficiency virus type 1 (HIV-1) from peripheral blood mononuclear cells (PBMC) requires 5 to 20 ml of blood, and the centrifugal separation of PBMC is expensive and time-consuming. Whole-blood coculture techniques use small sample volumes, do not require centrifugation, and allow measurement of the total viral burden in peripheral circulation. We compared the results of citrated whole-blood coculture with those obtained by the standard AIDS Clinical Trials Group PBMC semiquantitative culture method and reverse transcription-PCR quantitation of plasma HIV-1 RNA levels. PBMC cocultures were also set up with added erythrocytes (RBCs) to determine if the presence of RBCs affects the replication of HIV-1 in vitro. The mean number of cells required for a p24-positive PBMC coculture was approximately seven times greater than that required for a positive citrated whole-blood coculture (P < 0.01). At volumes of 100, 50, and 25 microl, the sensitivities of the whole-blood coculture were 94.5, 93.6, and 87.3%, respectively. The PBMC culture in the presence of added RBCs was more sensitive than PBMC coculture alone. The citrated whole-blood coculture was simple to perform, produced a reliable diagnosis of HIV infection in adult volunteers, was more sensitive than previously reported techniques even in half the culture time, and showed less variability than the PBMC coculture. Citrated whole-blood coculture may be a useful and efficient tool for diagnosing infection with HIV-1.

Non-invasive detection of ibuprofen in vivo 13C-NMR signals in rats.

A non-invasive in vivo 13C-NMR experiment is performed to follow the metabolic pathway of ibuprofen in rats. Detection of possible intermediates and the stereoselectivity of ibuprofen chiral inversion process are discussed.

[Image analysis on pulmonary arterioles in chronic cor pulmonale].

Thickness and area of the vessel wall and caliber and area of the vessel lumen of pulmonary arterioles obtained from 13 cases of chronic cor pulmonale were analysed. The relative ratios of vessel lumen area/total area(EA/TA), vessel wall area/total area(WA/TA) and vessel wall thickness/vessel external diameter(WD/TD) were calculated with image analysor. The results showed that compared with the pulmonary arterioles in patients without cor pulmonale, the relative ratios of EA/TA decreased significantly, while the values of WA/TA and WD/TD increased significantly in those with cor pulmonale owing to chronic bronchitis and diffuse interstitial fibrosis. The results obtained are of meaning for quantitative study and diagnosis about the pathology of cor pulmonale.