Improving the Accuracy and Efficiency of Abnormal Cervical Squamous Cell Detection With Cytologist-in-the-Loop Artificial Intelligence.

Population-based cervical cytology screening techniques are demanding and laborious and have relatively poor diagnostic accuracy. In this study, we present a cytologist-in-the-loop artificial intelligence (CITL-AI) system to improve the accuracy and efficiency of abnormal cervical squamous cell detection in cervical cancer screening. The artificial intelligence (AI) system was developed using 8000 digitalized whole slide images, including 5713 negative and 2287 positive cases. External validation was performed using an independent, multicenter, real-world data set of 3514 women, who were screened for cervical cancer between 2021 and 2022. Each slide was assessed using the AI system, which generated risk scores. These scores were then used to optimize the triaging of true negative cases. The remaining slides were interpreted by cytologists who had varying degrees of experience and were categorized as either junior or senior specialists. Stand-alone AI had a sensitivity of 89.4% and a specificity of 66.4%. These data points were used to establish the lowest AI-based risk score (ie, 0.35) to optimize the triage configuration. A total of 1319 slides were triaged without missing any abnormal squamous cases. This also reduced the cytology workload by 37.5%. Reader analysis found CITL-AI had superior sensitivity and specificity compared with junior cytologists (81.6% vs 53.1% and 78.9% vs 66.2%, respectively; both with P < .001). For senior cytologists, CITL-AI specificity increased slightly from 89.9% to 91.5% (P = .029); however, sensitivity did not significantly increase (P = .450). Therefore, CITL-AI could reduce cytologists' workload by more than one-third while simultaneously improving diagnostic accuracy, especially compared with less experienced cytologists. This approach could improve the accuracy and efficiency of abnormal cervical squamous cell detection in cervical cancer screening programs worldwide.

Protein Phosphatase 1 Regulatory Subunit 3: A Prognostic Biomarker in Stomach Adenocarcinoma.

PURPOSE: This study aimed to determine the potential application of the protein phosphatase 1 regulatory subunit 3 (PPP1R3B) gene as a prognostic marker in stomach adenocarcinoma (STAD), as well as its potential mediating biological processes and pathways. MATERIALS AND METHODS: Differential expression analyses were performed using the TIMER2.0 and UALCAN databases. Complete RNA-seq data and other relevant clinical and survival data were acquired from The Cancer Genome Atlas (TCGA). Univariate survival analyses, Cox regression modelling, and Kaplan-Meier curves were implemented to investigate the associations between PPP1R3B gene expression and clinical pathologic features. A genome wide gene set enrichment analysis (GSEA) was conducted to define the underlying molecular mechanisms mediating the observed associations between the PPP1R3B gene and STAD development. RESULTS: We found that PPP1R3B was overexpressed in STAD tissues, and that higher PPP1R3B expression correlated with worse prognoses in patients with STAD. Comprehensive survival analyses suggested that PPP1R3B might be an independent predictive factor for survival time in patients with STAD. The prognostic relationship between PPP1R3B and STAD was also verified using Kaplan-Meier curves. Patients with higher PPP1R3B levels had a shorter clinical survival time on average. Additionally, a GSEA demonstrated that PPP1R3B might be involved in multiple biological processes and pathways. CONCLUSION: Our findings demonstrate that the PPP1R3B gene has utility as a potential molecular marker for STAD prognoses.

The Impact of Health Investment on Industrial Structure: "Spillover" or "Crowding Out"?-Evidence From Emerging Market Countries.

Using national dynamic panel data from 21 emerging markets between 1999 and 2020 and bidirectional fixed effect and threshold regression methods, this paper evaluated the impact of health investment on industrial structure upgrading from two aspects of economic output and economic structure. The results showed that: (1) public health investment and private health investment have a crowding out effect on the added value of primary and secondary industries, and the crowding out effect of public health investment is greater than that of private health investment; (2) Public health investment and private health investment have a spillover effect on the added value of the tertiary industry, and the spillover effect of public health investment is greater than that of private health investment; (3) Both public and private health investment contribute to the transfer of the labor force to the tertiary industry, and tests showed the baseline regression results were robust and reliable; (4) The relationship between health investment and industrial structure upgrading was non-linear. As per capital GDP increases, the inhibition effect of public health investment on industrial structure upgrading gradually weakens whilst the promotion effect of private health investment on industrial structure upgrading gradually increases. The results of this study clarify how health investment affects industrial structure, and offers new guidance for health investment policy formulation in emerging market countries.

Contrast-enhanced ultrasound for the preoperative assessment of laryngeal carcinoma: a preliminary study.

BACKGROUND: Contrast-enhanced ultrasound (CEUS) is considered an attractive imaging technique to evaluate tumor microcirculation. However, the validity of CEUS for assessing laryngeal carcinoma is unclear. PURPOSE: To compare the performance of CEUS with conventional US and contrast-enhanced computed tomography (CECT) in the diagnosis and preoperative T-staging of laryngeal carcinoma. MATERIAL AND METHODS: Forty-one consecutive patients with laryngeal carcinoma underwent conventional high-frequency US, CEUS, and CECT before surgery. The CEUS characteristics of laryngeal carcinoma were recorded. The imaging findings of CEUS and conventional US were compared with CECT findings and the postoperative pathological examination. RESULTS: CEUS showed hyperenhancement in 38 cases and isoenhancement in three cases. Homogeneous distribution of contrast agent was found in 20 cases and heterogeneous distribution in 21 cases, of which 16 cases showed local perfusion defects. In the enhanced phase, rapid entry was observed in 37 cases, synchronous entry was observed in two cases, and slow entry was observed in two cases. Rapid exit was observed in 25 cases and slow exit was observed in 16 cases. The pretherapeutic T-staging accuracy was not significantly different between conventional US, CEUS, and CECT (P ≥ 0.500). A high sensitivity and specificity were achieved by CEUS in the evaluation of involvement of thyroid cartilage. CONCLUSION: Compared with conventional US and CECT, CEUS has a reliable initial T-staging accuracy and diagnostic properties for detecting laryngeal cartilage invasion.

Combined Acoustic Radiation Force Impulse and Conventional Ultrasound in the Quantitative Assessment of Immunoglobulin a Nephropathy.

We investigated the value of combined acoustic radiation force impulse (ARFI) imaging and conventional ultrasound (US) in identifying renal histopathological fibrosis with immunoglobulin A nephropathy. A total of 146 patients with immunoglobulin A nephropathy, pathologically confirmed by renal biopsy were grouped according to Oxford classification and Katafuchi grading, were included in the test group, and 39 healthy volunteers were included in the control group. Receiver operating characteristic (ROC) curves were constructed to compare the diagnostic accuracy of ARFI, renal lengths, parenchymal thicknesses and interlobular arterial resistance index (RI) and their combinations in identifying Katafuchi grading at renal biopsy. Shear wave velocity (SWV), renal length, renal parenchyma thickness and the interlobular arterial RI were correlated with Katafuchi grading, mesangial hypercellularity (M) and tubular atrophy/interstitial fibrosis (T) (r = -0.504 to -0.407, p < 0.01) but were not correlated with endocapillary hypercellularity (E) or segmental glomerulosclerosis (S). The area under the curves of SWV value + conventional US index (renal length, renal parenchyma thickness and interlobular arterial RI) was higher than those of the SWV value or of the conventional US index alone. The combination of ARFI imaging and conventional US can improve the diagnostic performance in quantitative evaluation pathologic damage in patients with immunoglobulin A nephropathy.

Value of Contrast-Enhanced Ultrasound and Acoustic Radiation Force Impulse Imaging for the Differential Diagnosis of Benign and Malignant Thyroid Nodules.

Objectives: To assess the value of contrast-enhanced ultrasound (CEUS) and acoustic radiation force impulse (ARFI) imaging for the differential diagnosis of benign and malignant thyroid nodules. Methods: CEUS was performed in eighty-eight thyroid nodules. The patterns of CEUS were analyzed, and ARFI was then performed. The shear wave velocities (SWVs) of the nodules and the surrounding normal thyroid tissue were obtained. The areas under the curve (AUCs) and cut-off value were obtained by a receiver operating characteristic (ROC) curve analysis. The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and diagnostic rate of each modality were assessed and compared using pathological diagnosis. Results: Among 88 nodules, 29 nodules were malignant and 59 were benign. The sensitivity, specificity, PPV, NPV, and diagnostic rate of CEUS were 79.3, 91.5, 82.1, 90, and 87.5%, respectively. Using a cut-off value of 2.565 m/s for SWV, the sensitivity, specificity, PPV, NPV and diagnostic rate for malignancy were 75.9, 94.9, 88.0, 88.9, and 88.6%, respectively. The AUC was 0.878. The sensitivity, specificity, PPV, NPV and diagnostic rate of CEUS in combination with ARFI were 93.1, 89.8, 81.8, 96.3, and 90.9%, respectively. Conclusion: Both CEUS and ARFI are valuable for the differential diagnosis of benign and malignant thyroid nodules. Combining these two methods can improve the diagnostic rate.

Gastrointestinal involvement by mantle cell lymphoma identified by biopsy performed during endoscopy: A case report.

RATIONALE: Primary gastrointestinal mantle cell lymphoma is rare, and histopathological examination and specific immunohistochemical staining are still the gold standard for diagnosis. Therefore, it is necessary to find a new way to improve positive biopsy rates. PATIENT CONCERNS: A 58-year-old man was admitted to our hospital with epigastric pain, abdominal distension, nausea, and melena. Endoscopy identified submucosal neoplasms and diffuse gastrointestinal tract involvement including the esophagus. DIAGNOSES: A false-negative diagnosis was first determined by ordinary endoscopy. However, a large tissue biopsy was subsequently performed using endoscopic mucosal resection based on endoscopic ultrasonography (EUS). Pathological examination of the biopsy specimens taken from the lesions of the duodenum and rectum revealed diffuse lymphocytic proliferation and obscure nodular and small cleaved cells with irregularly shaped nuclei. Immunohistochemistry showed that the cells were positive for CyclinD1, BCL-2, CD20, CD21, and CD5; however, they were negative for CD3, CD6, CD10, and CD43. INTERVENTIONS: The patient refused to receive further treatment. OUTCOMES: Mantle cell lymphoma was conclusively diagnosed. CONCLUSIONS: EUS has an important role in the diagnosis and management of gastrointestinal submucosal tumors. Performing a pathological biopsy including EUS may be useful for identifying the unknown nature of tumors of the digestive tract.

The correlation between HER-2 expression and the CEUS and ARFI characteristics of breast cancer.

OBJECTIVE: The purpose of the present study was to explore the correlation between the contrast-enhanced ultrasound (CEUS) and acoustic radiation force impulse (ARFI) characteristics of breast cancers and the expression of human epidermal growth factor receptor 2 (HER-2). METHODS: HER-2 expression levels in the tumor masses of 167 clearly diagnosed cases of breast cancer were measured and analyzed. The enhancement features and time intensity curve (TIC) of CEUS and virtual touch tissue imaging (VTI) and virtual touch tissue quantification (VTQ) technology of ARFI were employed to analyze the relationship between HER-2 expression and the CEUS and ARFI characteristics of breast cancer. RESULTS: (1) Statistically significant differences in the distribution of the contrast agent, perforator blood flow, the overranging phenomenon and perfusion defects between the study groups with different HER-2 expression levels (P < 0.05) were observed on CEUS. In addition, statistically significant differences in the TIC peak time (PT), slope of the ascending branch (K) and area under the curve (AUC) were found in the groups expressing different levels of HER-2 (P < 0.05). In contrast, the degree of contrast agent enhancement and TIC peak intensity (PI) were found to be independent of the expression status of HER-2, as they were not statistically significant (P > 0.05). (2) Statistically significant differences in the VTQ results between the groups with different HER-2 expression levels were found (P < 0.05). However, no statistically significant differences in VTI image characteristics were detected between the groups expressing different levels of HER-2 (P > 0.05). CONCLUSION: A correlation was found between the CEUS and ARFI characteristics of breast cancer and HER-2 expression levels. This correlation was principally reflected in perfusion defects, perforator blood flow, PI, PT, K and VTQ.

[Docosahexaenoic acid inhibits aflatoxin B1-induced migration and invasion in hepatocellular carcinoma cells in vitro].

OBJECTIVE: To investigate the effect of docosahexaenoic acid (DHA) on invasiveness of aflatoxin B1 (AFB1)-induced hepatocellular carcinoma cells in vitro. METHODS: HepG2.2.15 cells were exposed to different concentrations of AFB1 and DHA plus AFB1. The cell migration and invasion were assessed using wound-healing and Transwell assay, and flow cytometry was used to analyze the cell cycle changes. The ultrastructural changes of the cells were observed by transmission electron microscopy. RESULTS: Compared with the control group, the cells exposed to2 µmol/L AFB1 showed obviously enhanced migration and invasion with decreased cell ratio in G1/G1 phase and increased cell ratio in G2/M phase but no changes in S phase cells; transmission electron microscopy revealed the presence of multiple nucleoli and significantly increased mitochondria and Golgi apparatus in the exposed cells. Compared with AFB1-exposed cells, the cells treated with DHA and AFB1 showed decreased migration and invasion abilities, and the G1/G1 phase cells increased and G2/M phase cells decreased significantly; ultrastructurally, the cells contained single nucleoli with decreased mitochondria and vacuolization occurred in the cytoplasm. CONCLUSION: DHA can significantly inhibit AFB1-induced enhancement of cell migration and invasion in hepatocellular carcinoma cells in vitro.

RGD-Targeted Ultrasound Contrast Agent for Longitudinal Assessment of Hep-2 Tumor Angiogenesis In Vivo.

OBJECTIVE: To prepare arginine-glycine-aspartate (RGD)-targeted ultrasound contrast microbubbles (MBs) and explore the feasibility of their use in assessing dynamic changes in αvß3 integrin expression in a murine model of tumor angiogenesis. METHODS: RGD peptides were conjugated to the surfaces of microbubbles via biotin-avidin linkage. Microbubbles bearing RADfK peptides were prepared as controls. The RGD-MBs were characterized using an Accusizer 780 and optical microscopy. The binding specificity of the RGD-MBs for ανß3-expressing endothelial cells (bEnd.3) was demonstrated in vitro by a competitive inhibition experiment. In an in vivo study, mice bearing tumors of three different stages were intravenously injected with RGD-MBs and subjected to targeted, contrast-enhanced, high-frequency ultrasound. Subsequently, tumors were harvested and sectioned for immunofluorescence analysis of ανß3 expression. RESULTS: The mean size of the RGD-MBs was 2.36 ± 1.7 µm. The RGD-MBs showed significantly higher adhesion levels to bEnd.3 cells compared to control MBs (P < 0.01). There was rarely binding of RGD-MBs to αvß3-negative MCF-7 cells. Adhesion of the RGD-MBs to the bEnd.3 cells was significantly inhibited following treatment with anti-alpha(v) antibodies. The quantitative acoustic video intensity for high-frequency, contrast-enhanced ultrasound imaging of subcutaneous human laryngeal carcinoma (Hep-2) tumor xenografts was significantly higher in small tumors (19.89 ± 2.49) than in medium tumors (11.25 ± 2.23) and large tumors (3.38 ± 0.67) (P < 0.01). CONCLUSIONS: RGD-MBs enable noninvasive in vivo visualization of changes in tumor angiogenesis during tumor growth in subcutaneous cancer xenografts.

Relation between qualitative and quantitative 3-dimensional ultrasound and ki-67 expression in breast cancer.

To investigate the relation between quantitative blood flow parameters on 3-dimensional (3D) color histogram, 3D ultrasound characteristics and Ki-67 expression in breast cancer. Three-dimensional ultrasound characteristics and histological classifications of 76 breast tumors in 75 confirmed cases were analyzed. Relations of tumor volume (V), vascularization index (VI), flow index (FI) and vascularization-flow index (VFI) on 3D color histogram to Ki-67 expression were studied by statistical methods. VI and VFI measurements of tumors in positive Ki-67 expression group were obviously increased compared with the negative expression group (P<0.05). V and FI measurements of positive expression group were higher than those of the negative expression group, but the difference was not significant (P>0.05). Cases showing positive expression of Ki-67 were more likely to have lymph node metastases (P<0.05), and Ki-67 expression positively correlated with histological classification (P<0.05). However, the two groups did not show significant differences in the findings of "sun-like symptom" (P>0.05). Qualitative and quantitative 3D ultrasound characteristics correlated with positive expression of Ki-67 in breast cancer. Quantitative analysis with 3D color histogram more accurately evaluates blood supply of breast tumors, providing references for predicting biological behaviors and prognosis of breast cancer.

Acoustic radiation force impulse imaging for non-invasive assessment of renal histopathology in chronic kidney disease.

OBJECTIVE: To investigate the stiffness values obtained by acoustic radiation force impulse (ARFI) quantification in assessing renal histological fibrosis of chronic kidney disease (CKD). METHODS: 163 patients with CKD and 32 healthy volunteers were enrolled between June 2013 and April 2014. ARFI quantification, given as shear wave velocity (SWV), was performed to measure renal parenchyma stiffness. Diagnostic performance of ARFI imaging and conventional ultrasound (US) were compared with histologic scores at renal biopsy. Intra- and inter-observer reliability of SWV measurement was analyzed. RESULTS: In CKD patients, SWV measurements correlated significantly with pathological parameters (r = -0.422--0.511, P<0.001), serum creatinine (r = -0.503, P<0.001), and glomerular filtration rate (r = 0.587, P<0.001). The mean SWV in kidneys with severely impaired (histologic score: ≥19 points) was significant lower than that mildly impaired (histologic score: ≤9 points), moderately impaired (histologic score: 10-18 points), and control groups (all P<0.001). Receiver operating characteristic (ROC) curves analyses indicated that the area under the ROC curve for the diagnosis of renal histological fibrosis using ARFI imaging was superior to these conventional US parameters. Using the optimal cut-off value of 2.65 m/s for the diagnosis of mildly impaired kidneys, 2.50 m/s for moderately impaired kidneys, and 2.33 m/s for severely impaired kidneys, the corresponding area under the ROC curves were 0.735, 0.744, and 0.895, respectively. Intra- and intre-observer agreement of SWV measurements were 0.709 (95% CI: 0.390-0.859, P<0.001) and 0.627 (95% CI: 0.233-0.818, P = 0.004), respectively. CONCLUSIONS: ARFI may be an effective tool for evaluating renal histological fibrosis in CKD patients.

[Villous adenoma of the urinary tract: a clinicopathological study].

OBJECTIVE: To explore the clinicopathological features, immunophenotype, differential diagnosis, pathogenesis and prognosis of villous adenoma with poorly differentiated adenocarcinoma of the urinary tract. METHODS: Clinical and pathologic findings of 3 cases of villous adenoma with poorly differentiated adenocarcinoma of the urinary tract were analyzed by gross examination, microscopic investigation and immunohistochemical staining. The related literatures were reviewed. RESULTS: All of the three cases were middle-aged or elderly patients. Three cases all presented with hematuria and mucusuria. Endoscopic examination identified that case 1 had a polyp with broad attachment in the dome of bladder, case 2 had a solid mass in the ureter, and case 3 had a exophytic fungating tumor in the renal pelvis. Microscopically, case 1 revealed a papillary lesion with finger-like processes lined by pseudostratified columnar epithelium with abundant goblet cells. The cells demonstrated moderate degree dysplasia. In case 2 and case 3, both villous adenomas and poorly differentiated adenocarcinoma were observed, the adenoma cells arranged in a cribriform pattern, and the tumor cells showed severe atypia, mitotic activity, and transition with invasive poorly differentiated adenocarcinoma. Immunohistochemically, the tumor cells in three cases were positive for CK20, CEA,EMA and MUC-1; none of them expressed cdx-2 and PSA; In case 2 and 3, the same immunophenotype of villous adenomas and their associated adenocarcinomas was observed, but the number of the positive cells of p53 and Ki-67 staining were significantly increased in the area of adenocarcinomas than in that of the villous adenomas. CONCLUSIONS: Villous adenoma of the urinary tract is rare. It can occur in the urinary bladder, urachus, renal pelvis, ureter and urethra. These lesions may have malignant potential and frequently coexist with other malignant tumors. So, villous adenoma of the urinary tract should be removed completely and sampled thoroughly to avoid missing a more aggressive component.

[Plexiform angiomyxoid myofibroblastic tumor of stomach].

OBJECTIVE: To study the clinicopathologic features, immunophenotype and differential diagnosis of plexiform angiomyxoid myofibroblastic tumor (PAMT) of the stomach. METHODS: The clinical and pathologic findings of 3 cases of PAMT in the gastric antrum were retrospectively analyzed. Immunohistochemical study was carried out and the literature was reviewed. RESULTS: The age of patients ranged from 31 to 47 years. The male-to-female ratio was 1:2. The clinical presentation included epigastric pain and distension. Endoscopically, the tumor mass protruded into the gastric cavity at the antrum and ranged from 4.5 cm to 8.0 cm in greatest dimension. One of the tumors studied was associated with surface ulceration. Histologically, the tumors were located in the gastric wall. They were composed of bland spindle cells and small vessels arranged in a plexiform or nodular pattern within a myxoid stroma. Immunohistochemical study showed that the spindle cells were consistently positive for smooth muscle actin and muscle-specific actin. There was focal staining for h-caldesmon, desmin in case 3 and focal positive for epithelial membrane antigen, CAM5.2 in case 1. Further, CD10 and progesterone receptor were positive in case 3. CONCLUSIONS: PAMT represents a rare novel mesenchymal tumor of the stomach, with a propensity of gastric antral involvement. The distinctive pathologic features help to differentiate this entity from other benign and malignant tumors.

Increased susceptibility to experimental steatohepatitis induced by methionine-choline deficiency in HBs-Tg mice.

BACKGROUND: Worldwide, about 25% of individuals with chronic hepatitis B have fatty liver disease. Lipogenic diets that are completely devoid of methionine and choline induce nonalcoholic fatty liver disease. However, no animal model of nonalcoholic steatohepatitis associated with HBV infection is available, and the influence of viral infection on nutritional hepatic steatosis is unclear. METHODS: We used HBV surface antigen transgenic mice (HBs-Tg mice), which mimic healthy human carriers with hepatitis B surface antigen. The mice were fed with a high-fat methionine-choline-deficient diet (MCD) to build a reliable rodent nutritional model of nonalcoholic steatohepatitis associated with HBV infection, and the changes in body weight and serum triglycerides were measured. Hepatocyte ballooning changes were determined by hematoxylin and eosin staining. The extent of hepatic fat accumulation was evaluated by oil red O staining. Immunohistochemical assays were performed to detect proliferating cell nuclear antigen as an index of cell proliferation. RESULTS: MCD feeding provoked systemic weight loss and liver injury. MCD feeding caused more macrovesicular fat droplets and fat accumulation in the livers of HBs-Tg mice than in wild-type C57BL/6 mice. In addition, within 30 days of MCD exposure, more PCNA-positive nuclei were found in the livers of HBs-Tg mice. CONCLUSIONS: HBs-Tg mice fed with a lipogenic MCD form more macrovesicular fat droplets earlier, coincident with more hepatocyte proliferation, resulting in the appearance of increased susceptibility to experimental steatohepatitis in these mice.

Immunotherapeutical potential of Mycobacterium vaccae on M. tuberculosis infection in mice.

Tuberculosis remains the worldwide infectious disease. To identify the therapeutic potential of M. vaccae in treating tuberculosis, M. vaccae was injected into Mycobacterium tuberculosis (M. tuberculosis) infected mice. The optimal dose of M. vaccae (22.5 microg/mouse) treated mice showed lower pathological change index, spleen weight index, lung weight index and vital M. tuberculosis count than those of the untreated group. Treatment with M. vaccae enhanced the percentages of CD3+ and CD4+ T cells, IFN-gamma+CD4+ T cells, innate immune cells including NK cells, NK1.1+ T cells and gammadeltaT cells, and reduced the percentage of IL-4+CD4+ T cells. Therefore, M. vaccae could protect the mice from M. tuberculosis infection and improve mouse innate and adaptive cell-mediated immunity, suggesting that M. vaccae is a potential immunotherapeutic agent in pulmonary tuberculosis.

Application of optical tweezers in the research of molecular interaction between lymphocyte function associated antigen-1 and its monoclonal antibody.

Lymphocyte function associated antigen-1 (CD11a/CD18, LFA-1) plays an important role in the structure of the immunological synapse and is required for efficient lysis of cytotoxic T lymphocytes (CTLs) and natural killer (NK) cells. To study the activation mode of LFA-1 on the NK cell surface, optical tweezers were used in the work. As an emerging technology, optical tweezers are widely used to manipulate microscopic objects and measure the forces of molecular interactions in the field of biological research. In our study, a new platform was constructed to study the single molecular behavior of receptor on cell surface using optical tweezers. Based on the platform, the interaction between an NK cell and a polystyrene microsphere coated with anti-LFA-1 antibody was observed. The result confirmed that the adhesion forces between an NK cell and a polystyrene bead were time-dependent. According to our findings, we propose that anti-LFA-1 antibody may cause the clustering of LFA-1 on NK cell surface.

[Induction and mechanism study of anti-melanoma immune response by M. tuberculosis Ag85B].

AIM: To study the anti-melanoma immunity efficacy of Ag85B antigen gene therapy in vivo. METHODS: C57BL/6 mice were inoculated s.c. with B16 cells, and 8 days later the mice were inoculated s.c. again with B16 cells (control group 1), B16/pcDNA3 cells (control group 2) or B16/pcDNA3-Ag85B cells (experimental group), respectively. Tumor volume, survival time, serum IFN-gamma level and IL-4 level of 3 groups mice were observed. Antitumor activity of Ag85B was studied. RESULTS: From 12 to 23 day, the mean tumor volume of mice increased from 1.1058 cm(3) and 0.9123 cm(3) to 7.5983 cm(3) and 5.8746 cm(3) in the control group 1 and 2, respectively. But it increased from 0.5158 cm(3) to 1.5080 cm(3) in the experimental group. The mean survival time of mice was 24.1 days and 24.7 days in the control group 1 and 2, respectively. That was 27.8 days in the experimental group. Within 13 days after the last inoculation, the serum IFN-gamma level of all groups experienced increased (That increased to 26.3 ng/L, 23.0 ng/L and 25.2 ng/L in the control group 1, 2 and the experimental group, respectively). Subsequently, the serum IFN-gamma level in the two control groups decreased (That decreased to 19.3 ng/L and 18.3 ng/L in the control group 1 and 2) while it still augmented in the experimental group (That increased to 46.5 ng/L). IL-4 level was slightly but not significantly enhanced and then declined in all mice. CONCLUSION: Ag85B induced the increase of serum IFN-gamma level in the animals experiments, inhibited the tumor growth and prolonged the survival of the tumor-bearing mice.

Analysis tissue expression of IFN-gamma in IL-12 and/or IL-18 gene ablated naïve mice.

Interleukin 12 (IL-12) and/or interleukin 18 (IL-18) gene ablated mice were applied for the investigation of the tissue expression of interferon gamma (IFN-gamma). For IL-12(-/-), IL-18(-/-), IL-12(-/-)/18(-/-) and wt mice, reproductive performance were recorded and IFN-gamma concentrations in heart, lung, liver, spleen, kidney and serum were quantified by ELISA. There were no significant differences of IFN-gamma in heart, lung and kidney between 4 strains although control group was higher. It was observed that for IL-12(-/-) mice, compared with other 3 groups, IFN-gamma in liver and spleen were decreased (p < 0.05) and reproductive performance appeared to be impaired. Serum IFN-gamma level of IL-12(-/-)/18(-/-) mice was significantly higher (p < 0.05). It was showed that IFN-gamma productions under the normal condition were independent upon IL-12 and IL-18, its expressions in various tissues were different, and optimal IFN-gamma is necessary for the normal growth and development of mammals. This study is helpful for clinical cytokines therapy.

[Effect of natural killer cell line NK-92 against human ovarian carcinoma cells in vitro and in vivo].

OBJECTIVE: To study the cytotoxic activity of NK-92 cells irradiated against human ovarian cancer. METHODS: NK-92 cells were exposed to different doses of radiation and assayed for proliferation by a standard (3)H-thymidine incorporation assay and cell count by using trypan blue exclusion. The cytotoxic activity of NK-92 cells against targets was measured in a standard (51)Cr-release assay in vitro. The effectiveness of irradiated NK-92 cells on ovarian cancer was compared with the control group of cancers (without injection of irradiated NK-92 cells). RESULTS: (1) In vitro:The proliferation of NK-92 cells was inhibited by radiation of 4, 8 and 16 Gy, respectively. From the (3)H-thymidine incorporation data, irradiation by 4 Gy reduced cell proliferation to 29% of control, while 8 Gy reduced proliferation to 6%. The cytotoxicity of NK-92 cells at 4 Gy 2 days following irradiation was approximately 42%-62% for ovarian cancer cell HO-8910, while it was 33%-58% at 8 Gy. (2) In vivo: Tumor size in treatment group was (0.047 +/- 0.019) cm(3) on day 30 after inoculation, and (0.167 +/- 0.021) cm(3) on day 40 and (0.343 +/- 0.022) cm(3) on day 50, while the sizes were smaller in treatment group (P < 0.01). In addition, the tumor group animals died between 74-82 days after injection of HO-8910 cells, while the treatment group animals were alive over 120 days (P < 0.01). CONCLUSION: Our study indicates that injection of irradiated NK-92 cells may be a potentially effective treatment for human ovarian carcinoma.