Macrophages Promote Subtype Conversion and Endocrine Resistance in Breast Cancer.

BACKGROUND: The progression of tumors from less aggressive subtypes to more aggressive states during metastasis poses challenges for treatment strategies. Previous studies have revealed the molecular subtype conversion between primary and metastatic tumors in breast cancer (BC). However, the subtype conversion during lymph node metastasis (LNM) and the underlying mechanism remains unclear. METHODS: We compared clinical subtypes in paired primary tumors and positive lymph nodes (PLNs) in BC patients and further validated them in the mouse model. Bioinformatics analysis and macrophage-conditioned medium treatment were performed to investigate the role of macrophages in subtype conversion. RESULTS: During LNM, hormone receptors (HRs) were down-regulated, while HER2 was up-regulated, leading to the transformation of luminal A tumors towards luminal B tumors and from luminal B subtype towards HER2-enriched (HER2-E) subtype. The mouse model demonstrated the elevated levels of HER2 in PLN while retaining luminal characteristics. Among the various cells in the tumor microenvironment (TME), macrophages were the most clinically relevant in terms of prognosis. The treatment of a macrophage-conditioned medium further confirmed the downregulation of HR expression and upregulation of HER2 expression, inducing tamoxifen resistance. Through bioinformatics analysis, MNX1 was identified as a potential transcription factor governing the expression of HR and HER2. CONCLUSION: Our study revealed the HER2-E subtype conversion during LNM in BC. Macrophages were the crucial cell type in TME, inducing the downregulation of HR and upregulation of HER2, probably via MNX1. Targeting macrophages or MNX1 may provide new avenues for endocrine therapy and targeted treatment of BC patients with LNM.

An alkaloid from Menispermum dauricum, dauricine mediates Ca2+ influx and inhibits NF-κB pathway to protect chondrocytes from IL-1ß-induced inflammation and catabolism.

ETHNOPHARMACOLOGICAL RELEVANCE: Dauricine (DA) is a natural plant-derived alkaloid extracted from Menispermum dauricum. Menispermum dauricum has been used in traditional Chinese medicine as a classic remedy for rheumatoid arthropathy and is believed to be effective in alleviating swelling and pain in the limbs. AIM OF THE STUDY: Osteoarthritis (OA) is a classic degenerative disease involving chondrocyte death, and there is still a lack of effective therapeutic agents that can reverse the progression of the disease. Here we explored the therapeutic effects of DA against OA and further explored the mechanism. MATERIALS AND METHODS: The effect of DA on cell viability was assessed by CCK-8. IL-1ß-treated mouse chondrocytes were used as an in vitro model of OA, and apoptosis was detected by flow cytometry. QRT-PCR, western blotting, cell staining, and immunofluorescence were used to detect relevant inflammatory factors and cartilage-specific expression. RNA sequencing was used to identify pertinent signaling pathways. The therapeutic effect of DA was verified by micro-CT, histological analysis and immunohistochemical analysis in a mouse OA model. RESULTS: DA demonstrated a high safety profile on chondrocytes, significantly reversing the inflammatory response induced by IL-1ß, and promoting factors associated with cartilage regeneration. Moreover, DA exhibited a significant protective effect on the knee joints of mice undergoing ACLT-DMM, effectively preventing cartilage degeneration and subchondral bone tissue destruction. These positive therapeutic effects were achieved through the modulation of the NF-κB pathway and the Ca2+ signaling pathway by DA. CONCLUSION: Being derived from a traditional herb, DA exhibits remarkable therapeutic potential and safety in OA treatment, presenting a promising option for patients dealing with osteoarthritis.

Nanocarrier-based activation of necroptotic cell death potentiates cancer immunotherapy.

Even though immunological checkpoint inhibitors have demonstrated a potent anti-tumor effect in clinical practice, the low immunogenicity of the majority of tumors still results in a lower response rate and a higher resistance to mono-immunotherapy. Recent studies revealed that immunogenic cell death (ICD) augments T cell responses against some cancers, thus indicating that this combination therapy may further improve the anti-tumor immunity produced by anti-PD-1/PD-L1. Herein a robust synergetic strategy is reported to integrate the activation of necroptotic cell death and the subsequent using of immune checkpoint inhibitors. Liposomes have good biocompatibility and are widely used as drug carriers. Using liposomes as TNF-α-loaded nanoplatforms achieves in vivo tumor targeting and long-term retention in the tumor microenvironment. Tumor cells treated with TNF-α-loaded liposomes exhibited the hallmarks of ICD including the release of high mobility group box 1 (HMGB1) and lactate dehydrogenase (LDH). Additionally, the tumor cell necrosis caused by TNF-α induces the in situ release of tumor-specific antigens, thus increasing the dendritic cell (DC) activation and T cell infiltration when combined with the checkpoint blockade therapy. Collectively, significant tumor reduction is accomplishable by this synergetic strategy, in which TNF-α-loaded liposomes convert the tumor cell into an endogenous vaccine and improve the anti-tumor immunity of anti-PD-1/PD-L1.

Effect of Field Inoculation with Arbuscular Mycorrhizal Fungi on Rhizosphere Soil Structure of Paris polyphylla var. yunnanensis / 中国实验方剂学杂志

Objective:To investigate the effects of arbuscular mycorrhiza（AM） fungi inoculation on the rhizosphere soil of Paris polyphylla var. yunnanensis under field conditions，so as to provide a reference for the standardized cultivation and development of high-quality varieties of P. polyphylla var. yunnanensis. Method:The effects of inoculation with mycorrhizal fungi on the rhizosphere soil structure of P. polyphylla var. yunnanensis were observed through a combination of small-scale field inoculation test and laboratory analysis. Soil indexes were determined by soil agrochemical methods. Result:The treatment groups inoculated with exogenous AM fungi showed a regulatory effect on the infection rate and intensity of AM fungi infection in the root system of P. polyphylla var. yunnanensis. After treatment with AM fungi，the soil pH was basically not affected，and the contents of organic matter，available nitrogen，available phosphorus，and available potassium increased. Moreover，the number of fungi decreased，the number of bacteria and actinomycetes increased，and soil enzyme activities increased. The results of correlation analysis showed that there was a significant positive correlation among the soil physical and chemical indexes，especially the bacterial number and the three types of phosphatases showed extremely significant correlation （r=0.849，0.800，0.804，P<0.01）. Conclusion:The application of the two mixed fungicides could increase the number of microorganisms and enzyme activities in the rhizosphere of P. polyphylla var. yunnanensis， and there was a certain synergy effect among the soil factors. Among the three field trials，the effects in Anshun，Guizhou and Wanzhou，Chongqing were more ideal，which provided a theoretical and practical basis for large-scale promotion of P. polyphylla var. yunnanensis in the field.

Construction of chitosan/ZnO nanocomposite film by in situ precipitation.

In the present work, flexible chitosan/ZnO nanocomposite films were prepared by a green and facile method through in situ precipitation of nano-ZnO (nZnO) in the chitosan film. Zn(Ac)2 was added in chitosan solution to provide Zn2+, thus Zn2+ was fixed in the chitosan matrix and converted into nZnO through interaction with NaOH with heating. The structure and properties of the hybrid films were characterized by Field emission scanning electron microscope (FESEM), atomic force microscope (AFM), Fourier transform infra-red (FT-IR), X-ray diffraction (XRD) and tensile testing. The results indicated that there was strong coordination interaction existed between Zn2+ and chitosan matrix for the good dispersion of nZnO in the chitosan film. Furthermore, nZnO distributed evenly in the chitosan and aggregated to form micro-nano-binary hierarchical structure, mimicking lotus leaf structure. Therefore, this work provides an effective way to prepare biocompatible and antibacterial chitosan/ZnO nanocomposite films, showing potential applications in the fields of antibacterial packaging and dressings.

Hyaluronic acid-chitosan nanoparticles encoding CrmA attenuate interleukin-1ß induced inflammation in synoviocytes in vitro.

Osteoarthritis (OA) is a common degenerative joint disease characterized by inflammation of synoviocytes and degradation of cartilage. In the present study, hyaluronic acid/chitosan (HA/CS) nanoparticles were used as a vehicle for gene therapy of OA, and the cytokine response modifier A (CrmA) pDNA was proposed as the target gene. The HA/CS/pCrmA nanoparticles were prepared and the characteristics of the nanoparticles were examined. The nanoparticles were spherical, and the smallest size was obtained with the HA:CS weight ratio of 1:4. The release analysis exhibited a constant release over 29 days. The pDNA was completely combined with HA/CS nanoparticles and the HA/CS nanoparticles protected pDNA from degradation. Subsequently, rat synoviocytes were transfected with HA/CS/pDNA nanoparticles, and the results demonstrated that the HA/CS nanoparticles were able to improve the transfection capacity of pDNA. The cytotoxicity of the HA/CS/pDNA nanoparticles was additionally detected using a MTS assay to ensure that the HA/CS nanoparticle was a safe carrier. To additionally investigate the effects of HA/CS/pCrmA nanoparticles on synoviocytes in OA, the MMP­3 and MMP­13 gene expression levels were detected at the gene and protein expression levels. These results indicated that the HA/CS/pCrmA nanoparticles attenuated interleukin­1ß­mediated inflammation in synoviocytes. It was concluded that the HA/CS/pCrmA nanoparticles may provide a novel approach to the treatment of OA.

Spiral computed tomography evaluation of rabbit VX2 hepatic tumors treated with 20 kHz ultrasound and microbubbles.

The aim of the present study was to explore the therapeutic effect of 20 kHz ultrasound (US) and microbubbles (MBs) on rabbit VX2 liver tumors by spiral computed tomography (CT) scanning. A total of 16 New Zealand rabbits with hepatic VX2 tumors were divided into four groups: Control, MB, low-frequency US and US + MB. The treatment effect was evaluated by spiral CT scanning prior to, during and following treatment (at 0 weeks and the end of 1 and 2 weeks). The tumor growth rate was recorded. The specimens of VX2 tumors were collected for histological examination and transmission electron microscopy (TEM). No significant differences were observed between tumor areas measured by CT and pathology after 2-week treatment (P>0.05). The mean tumor growth rates in the control, MB, US and US + MB groups after 2 weeks of treatment were 385±21, 353±12, 302±14 and 154±9%, respectively (P<0.05, US + MB vs. the other three groups). Hematoxylin and eosin staining in the US + MB group revealed coagulation necrosis, interstitial hemorrhage and intravascular thrombosis. In the control, MB and US groups, tumor cells exhibited clear nuclear hyperchromatism. TEM of US + MB revealed vascular endothelial cell wall rupture, widened endothelial cell gaps, interstitial erythrocyte leakage and microvascular thrombosis, while intact vascular endothelial cells and normal erythrocytes in the tumor vessels were observed in the control, MB and US groups. A combination of 20 kHz US and MBs may effectively inhibit rabbit VX2 tumors. Spiral CT scanning is an ideal method to evaluate the US treatment on rabbit tumors.

New sonographic morphology score for the differentiation of malignant from benign ovarian tumors.

AIM: The aim of this study was to explore the efficacy of a new sonographic morphology score (SMS) to differentiate malignant from benign ovarian tumors. METHODS: Data on 84 ovarian tumors in patients hospitalized in our hospital between 2013 and 2014 were retrospectively analyzed. We established a new sonographic morphology score for ovarian tumors based on volume and structure scores. The efficacy of the new SMS was compared with that of Ueland's index (the old SMS). Receiver-operator curves (ROC) of the two SMS were constructed, and the areas under the curve were calculated and compared. The ROC of the new SMS was also compared with those for the patients' CA-125 and CA-72-4 levels. RESULTS: The area under the ROC of the new SMS for ovarian tumors was 0.836, while for the old SMS for ovarian tumors it was 0.709. By Z-test (Z = 2.452, P = 0.0384), there was a significant difference between the new SMS and the old SMS in the diagnosis of ovarian tumors. There was no significant difference in the area under the ROC between the new SMS and CA-125 and CA-72-4 in the diagnosis of ovarian tumors. With a cut-off value of 6, the sensitivity, specificity, positive prognostic value, and negative prognostic value of the new SMS were 0.797, 0.85, 0.944, and 0.567, respectively. CONCLUSION: The new SMS may be used to differentiate malignant ovarian carcinomas from benign tumors except in the case of ovarian thecoma.

The effects of percutaneous ethanol injection followed by 20-kHz ultrasound and microbubbles on rabbit hepatic tumors.

OBJECTIVE: Low-frequency ultrasound (US) in combination with microbubbles (MBs) is able to inhibit the growth of VX2 rabbit liver tumors. In this study, we investigated the feasibility of using percutaneous ethanol injection (PEI) followed by low-frequency ultrasound and microbubbles (USMB) to inhibit VX2 tumor growth. METHODS: Eighteen New Zealand rabbits with hepatic VX2 tumors were divided into three groups: PEI, low-frequency ultrasound and MBs followed by PEI (USMB + PEI), and PEI followed by USMB (PEI + USMB). PEI was performed by ultrasound-guided injection of 95% anhydrous alcohol into internal liver tumors in rabbits twice a week for 2 weeks. The US parameters were 20 kHz, 2 W/cm(2), 40% duty cycle, 5 min, and once every other day for 2 weeks. Magnetic resonance imaging (MRI) was used to observe tumors before and after treatment, to examine changes in the tumors, and to measure the diameters of the tumors. All animals were followed up for 180 days after tumor implantation. Autopsy was performed at the end of the scheduled follow-up or immediately after death. Anatomically observed metastatic sites included the liver, lung, abdomen, and pelvic cavity. The survival time of all rabbits was recorded. RESULTS: After 4 weeks of treatment, on MRI, the tumor diameters in the PEI, USMB + PEI, and PEI + USMB groups were 8.33 ± 1.83, 19 ± 2.61, and 4.5 ± 1.22 mm, respectively. There was a significant difference in tumor size indicated by MRI in the three groups. Tumor size was smaller in the PEI + USMB group than in the PEI and USMB + PEI groups, with t = 4.54, p = 0.0062, and t = 16.38, p < .0001, respectively. The PEI + USMB group showed the fewest metastasis sites (χ(2) = 11.7333, p = 0.0194) and the longest survival period (χ(2) = 7.448, p = 0.0241). CONCLUSION: Percutaneous ethanol injection followed by low-frequency ultrasound and microbubbles can be effective in inhibiting rabbit liver tumors and prolonging survival time.

Advances in the development of farnesoid X receptor antagonists / 中国药科大学学报

@#Farnesoid X receptor(FXR), a member of the nuclear receptor superfamily, plays an important role on the regulating bile acid, lipid and glucose homeostasis. FXR antagonist can stimulate the conversion of total cholesterol into bile acid by improving the activity of CYP7A1. It can also reduce the content of triglyceride, low-density lipoprotein and increase that of high-density lipoprotein by regulating the relevant target genes. Considering the above facts, FXR antagonist is expected to become a potential drug to treat hyperlipidemia. In recent years, great progress has been made in the research on FXR antagonists with their various structural types having been obtained. This article introduces natural and synthetic FXR antagonists reported in recent years.

MRI for differentiating ovarian endometrioid adenocarcinoma from high-grade serous adenocarcinoma.

PURPOSE: To investigate magnetic resonance imaging (MRI) features for differentiating ovarian endometrioid adenocarcinoma (OEC) from high-grade serous adenocarcinoma (HGSC). MATERIALS AND METHODS: Twenty-three patients with 25 OECs and 93 patients with 139 HGSCs confirmed by surgery and pathology underwent conventional MRI and diffusion-weighted imaging (DWI). The MRI features of the tumors, including laterality, size, shape, configuration, signal intensity, ADC value of solid component, enhancement, ascites, synchronous primary cancer (SPC) of the ovary and endometrium, and clinical stage, were evaluated and compared between two groups. RESULTS: The following characteristics were significantly more common for OECs than HGSCs: unilateral (91.3% vs 50.5%, P < 0.001), larger mass (80.0% vs 48.2%, P = 0.005), round or oval shape (64.0% vs 17.3%, P < 0.001), mainly cystic with mural nodules or papillary projections (72.0% vs 18.7%, P < 0.001), cystic component with homogeneous iso- or hyperintensity on T1WI (82.6% vs 4.3%, P < 0.001), moderate enhancement (52.0% vs 26.6%, P = 0.011), no or mild ascites (91.3% vs 57.0%, P = 0.002), and SPC (43.5% vs 4.3%, P < 0.001). The ADC value of the solid component was higher in OECs (0.979 ± 0.197 × 10(-3) mm(2)/s) than in HGSCs (0.820 ± 0.112 × 10(-3) mm(2)/s) (P = 0.002). When a mainly cystic mass with mural nodules or papillary projections was associated with any one of homogeneously iso- or hyperintense cystic component on TIWI, a relatively higher ADC value and SPC, the sensitivity, specificity, accuracy, and positive and negative predictive values for characterizing OEC were 87.0%, 93.5%, 92.2%, 76.9%, and 96.7%, respectively. CONCLUSIONS: Conventional MRI combining DWI is helpful for differentiating OECs from HGSCs.

Primary ovarian endometrioid adenocarcinoma: magnetic resonance imaging findings including a preliminary observation on diffusion-weighted imaging.

OBJECTIVE: This study aimed to investigate the magnetic resonance imaging (MRI) features of ovarian endometrioid adenocarcinoma (OEC) and to evaluate conventional MRI and diffusion-weighted imaging (DWI) for diagnosing OEC. MATERIALS AND METHODS: Twenty patients with OEC proven by surgery and pathology underwent MRI. The MRI features of the tumors evaluated included laterality, shape, size, configuration, mural nodules, signal intensity, apparent diffusion coefficient (ADC) values, enhancement, peritoneal implants, ascites, and synchronous primary cancer (SPC) of the ovary and endometrium. RESULTS: Unilateral ovarian masses were observed in 18 (90%) of the 20 patients with 22 OEC lesions, whereas the remaining 2 (10%) patients had bilateral masses. Oval, lobulated, and irregular shapes were observed in 13 (59%), 6 (27%), and 3 (14%) tumors, respectively. The maximum diameter of the tumors ranged from 3.7 to 22.5 cm, with a mean of 11.2 ± 5.1 cm. Fifteen (68%) masses were mainly cystic with mural nodules, 5 (23%) were mixed cystic-solid, and 2 (9%) were solid. The solid components of tumors showed isointensity (100%) on T1-weighted imaging (T1WI), heterogeneous hyperintensity on T2-weighted imaging (T2WI) (86%), and hyperintensity on DWI (82%), with a mean ADC value of (0.96 ± 0.20) × 10 mm/s. The cystic components showed isointensity or hyperintensity (85%) on T1WI, hyperintensity on T2WI (100%), and hypointensity on DWI (63%), with a mean ADC value of (2.27 ± 0.27) × 10 mm/s. Ten (50%) of the patients were SPC. The mean ADC values of the solid components were (0.85 ± 0.19) × 10 mm/s and (1.08 ± 0.15) × 10 mm/s in only-OEC and SPC, respectively, with a statistically significant difference (P = 0.012). CONCLUSIONS: Ovarian endometrioid adenocarcinoma usually appears as a large, oval, or lobulated cystic mass with mural nodules. Cystic components show isointensity or hyperintensity on T1WI, solid components and hyperintensity on T2WI and DWI. Synchronous primary cancer of the ovary endometrium is another characteristic feature of OEC.

Treatment of hepatic carcinoma by low-frequency ultrasound and microbubbles: A case report.

In vitro and in vivo studies have identified that low-frequency ultrasound (US) and microbubbles (MBs) mediate tumor inhibitory effects. However, the application of US in the clinical setting remains unclear. The aim of the present study was to investigate the clinically therapeutic effect of 20 kHz US in combination with MBs for the treatment of hepatic carcinoma. A 71-year-old male with a hepatic malignant tumor was admitted to Nantong University Affiliated Nantong Tumor Hospital (Nantong, China). The patient was subsequently sonicated with 20 kHz US and MBs over a period of five days. The low-frequency US parameters were set at 20 kHz, 2 W/cm2, duty cycle 40% (on 2 sec, off 3 sec) for a duration of 5 min each day for a total of five days. Computed tomography (CT), contrast-enhanced US (CEUS) and carbohydrate antigen 19-9 (CA19-9) tests were performed to evaluate the therapeutic effects. Although the tumor size increased marginally on CT from 5.4 to 5.6 cm after US treatment, the intensity and enhanced-areas on the CT scans and CEUS decreased. The abdominal lymph node decreased in size, from 2.2 to 1.9 cm, and CA19-9 levels decreased from the pretreatment value of 2,007 to 734 U/ml. Therapy with low-frequency US combined with MBs may exhibit an antivasculature effect and may be used as a palliative treatment for patients with unresectable hepatic malignant tumors.

Preoperative ultrasound features as prognostic factors for patients with hepatocellular carcinoma.

PURPOSE: Ultrasound is the most common imaging tool used to scan the tumours of hepatic carcinoma patients. However, very few studies have been performed to evaluate ultrasound imaging features for predicting tumour prognosis. Therefore, the goal of the current study was to evaluate preoperative ultrasound characteristics as prognostic factors that could affect survival rate after liver resection for hepatocellular carcinoma (HCC). MATERIALS AND METHODS: A total of 104 HCC patients who underwent resection were retrospectively reviewed with regard to their clinical data, preoperative ultrasound characteristics, and survival rate. Preoperative ultrasound parameters included cirrhosis, tumour site, size, echo pattern, portal vein thrombosis, intra-tumour blood flow signal, peak systolic velocity (V max), and resistance index (RI). The Kaplan-Meier method was used to calculate survival. Pre-resection prognostic factors were assessed using univariate log-rank test and a multivariate Cox proportional hazards model. RESULTS: The median survival was 37 months. The 1-, 3-, and 5-year disease-free survival (DFS) rates were 78.85, 53.85, and 26.92 %, respectively, and the overall survival (OS) rates at 1, 3, and 5 years were 85.58, 69.23, and 46.15 %, respectively. On univariate analysis, shorter survival was associated with mixed echo pattern, larger tumour size, portal vein thrombus, affluent flow signal, and higher V max. Application of the Cox multivariate proportional hazards model indicated that tumour size and blood flow signal in the tumours were independent prognostic factors. CONCLUSIONS: The overall survival for HCC patients undergoing hepatic resection can be stratified on a sonographic basis of tumour size and intra-nodular vasculature. These prognostic factors may be useful to determine appropriate treatment for HCC patients.

The effects of low-frequency ultrasound and microbubbles on rabbit hepatic tumors.

High-intensity focused ultrasound in combination with microbubbles (MBs) is able to inhibit the growth of VX2 rabbit liver tumors in vivo and prolong the survival time of the animals. In this study, we attempt to investigate the feasibility of VX2 tumor growth inhibition using low-frequency ultrasound (US)-mediated MB disruption. Forty-eight New Zealand rabbits with hepatic VX2 tumors were divided into four groups: control, MBs group, low-frequency US group, and US + MB group. The parameters of the US were 20 kHz, 2 W/cm², 40% duty cycle, 5 min, and once every other day for 2 weeks. At the end of the therapy experiment, 24 rabbits were euthanized, and the cancers were collected and cut into five sections for histological examination, immunohistochemistry, laser confocal microscopy, western blotting assays, and transmission electron microscopy (TEM). Another 24 rabbits were saved, and overall survival time was recorded. The tumor volumes in control, MB, US, and US + MB groups were 6.36 ± 0.58, 5.68 ± 0.42, 5.29 ± 0.26, and 2.04 ± 0.14 cm³, respectively (US + MB versus the other three groups, P < 0.01). Tumor cells manifested coagulation necrosis with internal calcification. Hematoxylin and eosin (H­E) staining revealed interstitial hemorrhage and intravascular thrombosis. The intensity of cyclooxygenase-2 (COX-2), and vascular endothelial growth factor (VEGF) in the US + MB group in the immunohistochemical staining, laser confocal microscopy, and western blotting assays was lower than that of the other three groups (P < 0.05). TEM of the US + MB group revealed vascular endothelial cell wall rupture, widened endothelial cell gaps, interstitial erythrocyte leakage, and microvascular thrombosis, while intact vascular endothelial cells and normal erythrocytes in the tumor vessels were observed in control, MB, and US groups. Rabbits treated with US + MB had a significantly longer overall survival than those in the other three groups (χ2 = 9.328, P = 0.0242). VX2 tumor growth could be inhibited by cavitation induced using low-frequency US and MB.

Computed tomography in predicting smear-negative pulmonary tuberculosis in AIDS patients.

BACKGROUND: The correct diagnosis of sputum smear-negative pulmonary tuberculosis in AIDS patients is very important to their therapy. We aimed to assess the value of the computed tomography (CT) and clinical findings in predicting smear-negative pulmonary tuberculosis in AIDS patients. METHODS: A total of 121 AIDS patients suspected of smear-negative pulmonary tuberculosis by clinical and radiographic findings were recruited. Pulmonary tuberculosis was diagnosed in 57 (47.1%) patients. The CT and clinical predictors were selected to diagnose AIDS-related pulmonary tuberculosis through univariate and multivariate Logistic analysis. RESULTS: Multivariate analysis showed that five variables, including weight loss, presence of miliary nodules, necrotic lymph node, lobular consolidation, tree-in-bud sign, were independent predictors of pulmonary tuberculosis in AIDS patients. Predicted scores based on the five variables were used to identify pulmonary tuberculosis. If the predicted score of 3 was taken as the ideal cut-off point in the diagnosis of AIDS-related smear-negative pulmonary tuberculosis, the sensitivity, specificity, positive predictive value, negative predictive value, and accuracy were 84.2%, 81.2%, 80.0%, 85.2%, and 82.6%, respectively. CONCLUSION: The prediction method based on five key factors of clinical and CT findings are useful in guiding the diagnosis of smear-negative pulmonary tuberculosis in AIDS patients.

The influence of T lymphocyte activation on HIV-1 susceptibility of Han Chinese / 中华预防医学杂志

<p><b>OBJECTIVE</b>To explore the influence of T lymphocyte activation on HIV-1 susceptibility of Han Chinese.</p><p><b>METHODS</b>In 2008, 37 HIV-1 highly exposed persistently seronegative individuals (ESNs) and 101 healthy controls were screened from Shenzhen. Flow cytometer was used to assay the expression difference of HIV-1 infection related co-receptor, the difference between the two groups were analyzed by Mann-Whitney U statistics methods.</p><p><b>RESULTS</b>T cell HLA-DR(+) CD4 T cells and HLA-DR(+) expression of ESNs (12.64 (5.94 - 21.90), 21.12 (10.74 - 30.21)) were all significantly lower than that of healthy controls (22.52 (7.91 - 58.60), 32.28 (14.72 - 67.82)) (P values all < 0.05). T cell CD45RA-RO(+), CCR5(+)CD4 expression of ESNs (58.68 (49.06 - 72.44), 21.93 (15.84 - 25.89)) were all significantly higher than that of healthy controls (53.17 (42.63 - 63.21), 16.14 (11.94 - 21.98)) (P values all < 0.05). T cell CXCR4(+)CD4 T cells expression of ESNs (93.67 (92.17 - 94.96)) was significantly lower than that of healthy controls (95.16 (92.99 - 96.77)) (P values all < 0.05). Healthy controls and ESNs could be divided into low expression group and high expression group according to HLA-DR(+)CD8 T cells bimodal distribution. A total of 89.2% (33/37) ESNs fell into HLA-DR + CD8 low expression group, and 58.4% (59/101) of the healthy controls located in low expression group (P < 0.05).</p><p><b>CONCLUSION</b>To Han Chinese, the low activation status of T lymphocyte has significant correlation with HIV-1 low susceptibility.</p>

A novel mutation Glu441stop (GAA to TAA) of androgen receptor gene resulting in complete androgen insensitivity syndrome / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To identify the mutation of human androgen receptor gene (AR) in a patient with complete androgen insensitivity syndrome (CAIS).</p><p><b>METHODS</b>DNA sequences of 8 exons and their exon/intron boundaries of the AR gene in the patient were amplified by PCR and directly sequenced.</p><p><b>RESULTS</b>DNA sequencing revealed a nonsense mutation in exon 1, resulting in a change of codon 441 GAA (glutamic acid) to a stop codon (TAA).</p><p><b>CONCLUSION</b>A novel mutation Glu441stop (GAA to TAA) of the androgen receptor gene leading to complete androgen insensitivity syndrome was identified in this study in a Chinese patient. It may help us further understanding the pathogenesis of CAIS.</p>

Molecular epidemiological survey on HIV-1 among the injecting drug users population in Shenzhen / 中华流行病学杂志

Objective To study the prevalent status of human immunodeficiency virus-1 (HIV-1) subtypes in IDU (injecting drug users) population in Shenzhen and to study their source of infection in order to predict the epidemic trend and evolution. Methods 166 HIV-1 positive plasma from the IDUs was collected from 1996 to 2008. HIV-1 env genes were amplified by nested-PCR from RNA. The C2-V3 regions (450 bp) of HIV-1 env were sequenced for analyses. Phylogenetic analyses were performed on the nucleotide sequence data. Results Among 166 samples, there were 6 HIV-1 strains including CRF01_AE, CRF08_BC, CRF07_BC 3 circulating recombinant forms (CRFs) and B',C, A1 3 subtypes. Data from the genotype analyses showed that 65.06% (108/166) were CRF01_AE, 19.88% (33/166) were CRF07 BC_6.02% (10/166) were CRF08_BC, 7.23%(9/166) were subtype B', 0.60% (1/166) were subtype C and 1.20% (2/166) were subtype A1. Phylogenetic tree analysis showed that some of HIV-1 clusters defined in CRF01_AE, CRF07_BC and subtype B' in different time groups. Significant increase of gene distance in CRF01_AE and CRF07_BC strains in the three different periods. Conclusion CRF01_AE and CRF07_BC were the major epidemic CRF strains among the IDU population in Shenzhen while the subtype B', C, A1 and CRF08_BC were also circulating in IDU population in this region. The variation of all different subtypes was increasing through these years.

Expression and activity analysis of human 67kD laminin receptor in Pichia pastoris / 生物工程学报

To carry out the secretive expression of human 67 kD laminin receptor (67LR), recombinant expression plasmid pPIC9K-67LR was constructed by inserting of 67LR cDNA into yeast expression vector pPIC9K. The 67LR protein was expressed in Pichia pastoris after induced by methanol, and about 12.56 mg electrophoresis purity 67LR could be obtained after the purification of 1L culture using affinity chromatograph column. In vitro competitive binding assay showed that target protein has an excellent biological activity. The successful expression of 67LR has placed a solid foundation for the research on structure and functions of 67LR.