[Hypersensitivity pneumonia diagnosed by excisional lung biopsy: a clinicopathological features of 47 cases].

Objective: To analyze the clinicopathological and immunohistochemical characteristics of the hypersensitivity pneumonia (HP) cases that were diagnosed by excisional lung biopsy, to improve the diagnosis accuracy of HP. Methods: The data of 47 HP cases diagnosed by excisional lung biopsy during the last 20 years were collected in Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, from January 2000 to June 2020. The clinicopathological features and the immunohistochemical profiles of CD3, CD4, CD8 and CD20 were analyzed using light microscopy and immunohistochemical stains. Results: The age of the patients ranged from 15 to 68 years, while the ratio of male to female was 22∶25. The main symptoms were shortness of breath and cough, and 32 cases had a history of allergen exposure. Most of the patients had restrictive impaired ventilatory function, and impaired diffusion function. The proportion of lymphocytes in bronchoalveolar lavage fluid (BALF) increased in 85.3%(29/34) of the patients, and the CD4+/CD8+ T cell ratio was not higher than 0.8 in 31.3%(10/32) of the patients. Microscopically, 59.6%(28/47) of the cases showed nonnecrotizing granuloma/giant cells in the stroma, and 40.4% (19/47) of the cases had only giant cells. Ninety-three-point six percent of the cases(44/47) had peribronchiolar metaplasia, while 70.2%(33/47) had focal cell-rich inflammation. Forty-four cases (93.6%, 44/47) had fibrosis, which was manifested as usual interstitial pneumonia-like fibrosis (47.7%, 21 cases), nonspecific interstitial pneumonia-like fibrosis (29.5%, 13 cases), purely peribronchiolar fibrosis (6.8%, 3 cases) and mixed fibrosis (15.9%, 7 cases). Immunohistochemistry showed that CD4+ T cells were fewer than CD8+ T cells in 24 cases (55.8%, 24/43) of HP. Thirty-two HP patients had follow-up data. Among them, 7 patients had improved symptoms, 12 patients were stable, and 13 patients had worsening symptoms. Conclusions: The most common type of HP diagnosed by excisional lung biopsy is chronic HP. The clinical characteristics are long course of disease, restricted ventilation disorder and impaired diffusion function with the increase of lymphocyte proportion in BALF. Nearly 1/3 of the patients have no known history of allergen contact. The degree of fibrosis in HP patients diagnosed by excisional lung biopsy is severe, and the histologic types of fibrosis varies, but the UIP-like and/or NSIP-like fibrosis is most common. The CD4+/CD8+ T cell ratio in lung tissues is less than 1 in more than half of the cases. Pulmonary fibrosis continued to increase in some of the patients with chronic HP, even after treatment.

[The 485th case: fever of undetermined origin and hypoxemia].

A 49-year-old male was admitted to Peking Union Medical College Hospital presented with fever for more than half a year. The patient was diagnosed as Sjogren's syndrome at local hospital. After oral prednisone 60 mg per day was given, the fever alleviated, but recurred after prednisone tapered to 40 mg/d. Both blood culture and stool culture were positive for Salmonella enteritidis. Antibiotics including ceftazidime, ceftriaxone, cilastatin-imipenem were sequentially administrated for 4 weeks, yet not effective. Although there were not respiratory symptoms or certain abnormalities on high-resolution chest CT, arterial blood gas indicated hypoxemia. Serum lactate dehydrogenase and ß2 micro-globulin were elevated, and the lung function test demonstrated significant impairment of diffusion function. Positron emission tomography-computed tomography (PET/CT)scan suggested that high fluorodeoxyglucose uptake was diffusely seen in both lungs. The patient was finally diagnosed as pulmonary intravascular large B-cell lymphoma (IVLBCL) by transbronchial lung biopsy. This case aims to emphasize the differentiation diagnoses of pulmonary intravascular lymphoma from common situations.

[The spectrum of pathogens in 187 cases of pulmonary fungal disease diagnosed by histopathology-a retrospective analysis].

Objective: To investigate the spectrum of pathogens causing lung fungal disease diagnosed by histopathology through histochemical special staining, compared to the fungal culture results, and to further evaluate the diagnostic value of histochemical special staining in pulmonary fungal disease. Methods: We performed a retrospective analysis of 187 cases of pulmonary fungal disease diagnosed by histopathology in Peking Union Medical College Hospital from 2001 to 2015 (including 92 cases with pulmonary resection or open lung biopsy, 33 with percutaneous lung biopsy and 62 ones with fiberoptic bronchoscopic lung biopsy). All cases were treated with hexamine silver, PAS, mucus carmine and acid-fast staining in addition to conventional HE staining. The clinical records and the fungal culture results were reviewed. Results: There were 103 male and 84 female patients, aged from 12 to 70 years [average (48±14) years]. There were 85 cases(45.5%) of pulmonary aspergillosis(including 60 cases of invasive infection and 25 cases of aspergilloma), 51 cases(27.3%) of pulmonary cryptococosis, 6 cases (3.2%)of pulmonary mucormycosis, 3 cases(1.6%) of pulmonary histoplasmosis, 3 cases (1.6%)of pulmonary candidiasis, and 2 cases (1.1%) of pneumocystosis, while in the remaining 37 cases (19.8%) the pathogens could not be clearly classified by microscopy due to limited tissue or degeneration. Among the 88 patients with pulmonary fungal disease diagnosed by histopathology from 2011 to 2015, 35 ones (39.9%) were detected by fungal culture (including lung biopsy, intraoperative swab, blood, bronchoalveolar lavage fluid and sputum, etc.). The diagnostic results of 18 cases were completely consistent between histopathological examination and fungal culture (18/35, 51.4%), while 13 cases (13/35, 37.1%) were diagnosed by histopathology but no fungi were cultured, and in 3 cases (3/35,8.6%) the culture was positive for fungi which could not be classified clearly by histopathology. In another case the pathogen was found to be Cryptococcus histopathologically but the lavage culture grew"candida", but the patient's blood cryptococcal antigen was positive. Conclusions: Among patients with histopathological diagnosis of pulmonary fungal disease, pulmonary aspergillosis was the most common, followed by pulmonary cryptococcosis, pulmonary mucormycosis, pulmonary histoplasmosis, pulmonary candidiasis and pneumocystosis. A small number of cases could not be classified by histopathology through histochemical special staining. There was a high consistency in discovering fungal pathogens between pathological histochemical special staining and culture method, but 37% pulmonary fungal disease diagnosed by histopathology were culture negative. In practice, the role of histochemical special staining in diagnosing pulmonary fungal disease should be paid more attention.

[Primary carcinoid of ovary: a clinicopathologic analysis of 17 cases].

Objective: To study the clinicopathologic feature, diagnostic strategy and prognostic significance of primary carcinoid of the ovary (PCOTO). Methods: A series of 17 patients previously diagnosed as PCOTO at Department of Pathology, Peking Union Medical College Hospital during the period from 2002 to 2017 were evaluated with clinical data analysis, histopathology and immunohistochemistry, and the patients were followed up and the relevant literatures were reviewed. Results: The age of patients ranged from 24 to 64 years (mean, 42 years). Fourteen patients were found a pelvic mass for a health check-up, and only 3 patients presented with pain in the lower abdomen. The left ovary was involved initially in ten patients, and the right in seven. The major axis of the tumor ranged from 2 to 14 cm. The surface of these tumors was smooth. Seven of the tumors were solid-cystic, 6 were cystic and 4 were solid. Under light microscope, 6 cases were diagnosed as strumal carcinoid, 4 cases were insular carcinoid, 4 cases were trabecular carcinoid, 3 cases were insular and trabecular mixed type carcinoid and 1 case was mucinous carcinoid. The mitotic figures were no more than 1/10 HPF.There were 11 cases complicated with other ovarian tumors, including 10 cases with teratoma, and 1 case with mucinous cystic adenoma. The paraffin-embedded tissues of all cases showed immunoreactivity for NSE and Syn, and the positive propotion of CgA was 10/17. TTF1 was positive in thyroid follicles and negative in strumal carcinoid. The positive index of Ki-67 was no more than 2%. Follow-up of 13 to 188 months showed 16 patients without recurrence and 1 patients were loss to follow-up. Conclusions: PCOTO is very rare. Most of the patients are found with a mass during health check-up in unilateral ovary and without obvious clinical symptoms. Histologically, the tumor always exists with other ovarian tumors, including teratoma and mucinous cystic adenoma, with relatively good prognosis.

Molecular cloning of heat shock protein 10 (Hsp10) and 60 (Hsp60) cDNAs from Galeruca daurica (Coleoptera: Chrysomelidae) and their expression analysis.

Galeruca daurica (Joannis) is a new outbreak pest in the Inner Mongolia grasslands in northern China. Heat shock protein 10 and 60 (Hsp10 and Hsp60) genes of G. daurica, designated as GdHsp10 and GdHsp60, were cloned by rapid amplification of cDNA ends techniques. Sequence analysis showed that GdHsp10 and GdHsp60 encoded polypeptides of 104 and 573 amino acids, respectively. Sequence alignment and phylogenetic analysis clearly revealed that the amino acids of GdHsp10 and GdHsp60 had high homology and were clustered with other Hsp10 and Hsp60 genes in insects which are highly relative with G. daurica based on morphologic taxonomy. The mRNA expression analysis by real-time PCR revealed that GdHsp10 and GdHsp60 were expressed at all development stages and in all tissues examined, but expressed highest in eggs and in adults' abdomen; both heat and cold stresses could induce mRNA expression of GdHsp10 and GdHsp60 in the 2nd instar larvae; the two Hsp genes were expressed from high to low with the extension of treatment time in G. daurica eggs exposed to freezing point. Overall, our study provides useful information to understand temperature stress responses of Hsp60 and Hsp10 in G. daurica, and provides a basis to further study functions of Hsp60/Hsp10 relative to thermotolerance and cold hardiness mechanism.

[Analysis of related factors on effects of uterine artery embolization in the treatment of dysmenorrhea of adenomyosis and the construction and validation of prediction model].

Objective: To investigate the related factors on effects of uterine artery embolization(UAE)in the treatment of dysmenorrhea in patients with adenomyosis, and to construct and validate the efficacy prediction model. Methods: A total of 127 cases of adenomyosis patients with symptoms of dysmenorrhea in Guangzhou No.1 People's Hospital and Nanfang Hospital of Southern Medical University from June 1999 to December 2009 were reviewed. The evaluation standard was to improve the degree of dysmenorrhea, the related factors of efficacy were analysed. Combined with artificial neural network theory, the effect prediction model was constructed, and the effectiveness of the model was evaluated using receiver operating characteristic(ROC)curve, and the effectiveness of the cut-off point was calculated. The model was validated by 68 cases of patients with adenomyosis in the Nanfang Hospital from January 2010 to November 2014. Results: (1)In 127 cases of dysmenorrhea patients, UAE treatment was effective in 98 cases, effective rate was 77.2%(98/127).(2)Age was an independent predictor of effective UAE treatment(HR= 1.129, P=0.026); in the range of this study, the greater the age, the higher the UAE treatment efficiency.(3)The developing situation of ovary branches of uterine artery was an independent predictor of effective UAE treatment(HR=0.460, P=0.020), the efficiency of patients whose intraoperative bilateral uterine artery ovarian branch did not develop was 89.7%(35/39), the efficiency of patients whose unilateral uterine artery ovarian branch was developing was 84.1%(37/44)and the efficiency of patients whose bilateral uterine artery ovarian branch were developing was 59.1%(26/44).(4)Blood supply of adenomyosisis was an independent predictor of effective UAE treatment(HR=0.313, P=0.001). Type Ⅰ(bilateral predominated)patients, efficiency was 93.5%(43/46); type Ⅱ(bilateral balanced)patients, efficiency was 78.0%(39/50); type Ⅲ(unilateral predominated)patients, efficiency was 51.6%(16/31).(5)UAE for the treatment of adenomyosis efficacy of artificial neural network prediction model was constructed, the model's area under the ROC curve was 0.808, the optimal cut-off point was 0.669 13. Actual verification of the model, sensitivity was 96.5%, specificity was 81.8%, positive predictive value was 96.5% and negative predictive value was 81.8%, the total accuracy was 94.1%. Conclusions: (1)Age, the developing situation of ovary branches and blood supply of adenomyosis are the independent predictors of effective UAE treatment.(2)The artificial neural network prediction model is satisfied with the accuracy and the accuracy of prediction.

[Analysis of the effect of cochlear implant for the patients with acoustic neuroma in the only hearing ear].

OBJECTIVE: To discuss the method as well as effect of hearing restoration for patients with acoustic neuroma (AN) in the only hearing ear. METHODS: A retrospective study was conducted in three patients with AN in the only hearing ear. Ipsilateral cochlear implantation (CI) was performed in case 1 and contralateral CI was performed in case 2 without tumor resection. In case 3, CI was performed 6 months after tumor removal by the retrosigmoid approach. Dynamic changes in the audiological tests was recorded regularly. RESULTS: In case 1 and case 2, no matter which side CI was performed, pure tone average (PTA) was improved clearly, and speech discrimination score (SDS) was slightly increased. In case 3, hearing loss was observed after tumor removal. Simultaneous bilateral CI after 12 months was performed in this case. PTA on both sides was largely lower than that before CI, and SDS was slightly improved. With bilateral CI, SDS was improved, especially in noise, compared with unilateral CI on either side. CONCLUSIONS: For patients with AN in the only hearing ear, CI can significantly improve PTA, and SDS can be slightly increased. Bilateral CI should improve the outcome of hearing restoration compared with unilateral CI.

[Effect of autophagy inhibitor combined with EGFR inhibitor on triple-negative breast cancer MDA-MB-468 and MDA-MB-231 cells].

OBJECTIVE: To investigate the effect of combined administration of autophagy inhibitor 3-methyladenine/bafilomycin A1 and EGFR inhibitor gefitinib on triple-negative breast cancer MDA-MB-468, MDA-MB-231 cells and estrogen receptor-positive MCF-7 cells. METHODS: All the cells were treated with 3-methyladenine/bafilomycin A1 and/or gefitinib. The effect of autophagy inhibitor and gefitinib on the cell growth was evaluated by MTT assay. Cell apoptosis was detected by flow cytometry. Western blot analysis was used to determine the alteration of autophagy-related protein (such as LC3) and apoptosis-related proteins (such as caspase-3 and caspase-9). RESULTS: MTT assay showed that the IC50 in the GE+ 3-MA and GE+ BAF groups were (4.1±0.2) µmol/L and (3.8±0.3) µmol/L, significantly lower than that of the gefitinib alone group [(7.0±0.2) µmol/L] in MDA-MB-468 cells (P<0.05). Similarly, the IC50 in the GE+ 3-MA and GE+ BAF groups were (9.7±0.1) µmol/L and (7.7±0.2) µmol/L, significantly lower than that of the gefitinib alone group [(14.7±0.1) µmol/L]in MDA-MB231 cells (P<0.05). The flow cytometry assay revealed that the apoptosis rates of MDA-MB-468 cells in GE, GE+ 3-MA and GE+ BAF groups were (12.43±3.18)%, (23.37±2.71)% and (18.71±2.81)%, respectively. The apoptosis rates of MDA-MB-231 cells of the GE, GE+ 3-MA and GE+ BAF groups were (12.15±1.82)%, (16.94±2.19)% and (33.83±5.92) %, significantly higher than that of the gefitinib alone group (All P<0.05). The apoptosis rates of the MCF-7 cells were not changed significantly among the three groups (P>0.05). Western blot data showed that the expression levels of LC3 and p-Akt were decreased in the combined groups than that of the gefitinib alone group, while the p-PTEN, caspase-3 and caspase-9 were increased. CONCLUSIONS: Autophagy inhibitor may enhance the sensitivity to gefitinib in MDA-MB-468 and MDA-MB-231 cells by activation of the PTEN/P13K/Akt pathway. Apoptosis in MDA-MB-468 and MDA-MB-231 cells might be enhanced by the combination treatment through caspase cascade.

Androgen receptor (CAG)n polymorphisms and breast cancer risk in a Han Chinese population.

The androgen receptor (AR) is involved in the differentiation and growth of breast cancer. Genetic markers in the AR gene have a plausible role in modulating the risk of breast cancer. In this study, we studied the association of breast cancer and the trinucleotide repeat polymorphism (CAG)n in exon 1 of the AR gene in 202 patients with breast cancer and 183 healthy controls from our hospital (Yinchuan, China). Repeat lengths were determined by fluorescent DNA fragment analysis using the ABI GeneScan software and DNA sequencing. We detected 17 short tandem repeat alleles in exon 1 in the Han population of Ningxia Province, China. The CAG repeat number ranged from 14 to 31 and the frequency ranged from 0.339 to 24.460%. Generally, (CAG)n repeat lengths <22 were classified as short (S), and those >22 were classified as long (L). No association was found between breast cancer and the S/L (CAG) variants. However, the frequency of the (CAG)25 repeats in the breast cancer group was significantly higher than that in the control group (P = 0.033, odds ratio = 1.790, 95% confidence interval = 1.044-3.069). These findings indicate a role for AR gene (CAG)n variations in breast cancer and might be informative for future genetic or biological studies on breast cancer, although these findings need replication in other populations.

Functional polymorphisms of the glutamate receptor N-methyl D-aspartate 2A gene are associated with heroin addiction.

Heroin dependence is a debilitating psychiatric disorder with a complex inheritance mechanism. Genetic polymorphisms in functional regions of the glutamate receptor, N-methyl D-aspartate 2A (GRIN2A) gene, which encodes the 2A subunit of the N-methyl D-aspartate (NMDA) receptor, may modulate the risk of heroin addiction. We investigated the potential association between 8 single nucleotide polymorphisms (SNPs) of the GRIN2A gene (SNPs rs3219790, rs1014531, rs8044472, rs8045712, rs9933624, rs9940680, rs1420040, and rs767749) and heroin addiction using the MassARRAY system and GeneScan. A total of 405 heroin-addicted patients and 397 healthy control subjects were recruited for this study. Statistically significant differences were observed for rs3219790 in the promoter region of the GRIN2A gene. The frequency of the (GT)26 repeats in the heroin addiction group was significantly higher than that in the control group [X(2) = 5.475, P = 0.019, odds ratio (OR) = 1.367, 95% confidence interval (CI) = 1.051-1.776]. Strong linkage disequilibrium was observed in block 1 (D' > 0.9). However, significant evidence of linkage disequilibrium was not observed between the 7 SNPs in our sample population. These data suggest that GRIN2A gene polymorphisms confer susceptibility to heroin addiction and support the hypothesis that dysfunction of GRIN2A is involved in the pathophysiological process of heroin addiction.

Meta-analysis: E-cadherin immunoexpression as a potential prognosis biomarker related to gastric cancer metastasis in Asian patients.

OBJECTIVE: The prognostic potential of reduced E-cadherin expression is associated with an increased risk of gastric cancer. However, its role in gastric cancer remains poorly understood. This study was to quantitatively summarize available evidences for evaluating E-cadherin immunoexpression in Asian patients with gastric cancer as a prognostic indicator. MATERIALS AND METHODS: Searches were applied to MEDLINE, EMBASE, the Cochrane Library and Chinese Biomedicine Databases until June 2012, without language restrictions. Studies were pooled and summary risk ratio (RR) or odds ratio (OR) were calculated. Potential sources of heterogeneity were sought out via subgroup and sensitivity analyses, and publication bias were also conducted. RESULTS: Our combined results showed that reduction of E-cadherin expression in Asian patients with gastric cancer was frequently observed as compared to the counterpart normal tissue (odds ratio [OR] = 64.16, 95% confidence interval [CI] = 24.53-167.80, p < 0.001). All the analyses estimated favored a stronger link between the reduced E-cadherin expression and the poor 5 year overall survival (risk ratio [RR] = 1.50, 95% CI = 1.36-1.66, p < 0.001). When stratifying the studies by the clinical variables, the depth of invasion (OR = 2.46, 95% CI = 1.70-3.57, p < 0.001), lymph node spread (OR = 1.83, 95% CI = 1.49-2.26, p < 0.001), distant metastasis (OR = 2.04, 95% CI = 1.45-2.87, p < 0.000), and TNM stage (OR = 2.11, 95% CI = 1.58-2.83, p < 0.001) provided significant prognostic information. CONCLUSIONS: Our findings indicate that E-cadherin appears to predict the overall survival and mark metastasis in Asian patients with gastric cancer. Importantly, E-cadherin may be implemented in the routine clinical management of gastric cancer. However, further pursuit of this possibility is warranted.

Roles of functional NFKB1 and ß-TrCP insertion/deletion polymorphisms in mRNA expression and epithelial ovarian cancer susceptibility.

Epithelial ovarian cancer (EOC) is the leading cause of death among all gynecological cancers. Nuclear factor-kappa B (NF-κB) is involved in carcinogenesis and in the development of EOC. The ß-transducin repeat-containing protein (ß-TrCP) is a positive regulator of the NF-κB signaling pathway. Recent studies have indicated that the -94 ins/del ATTG polymorphism in the promoter region of the NFKB1 gene, and the 9N ins/del polymorphism in the 3'-untranslated region of the ß-TrCP gene are associated with increased susceptibility to a variety of cancers. We examined a potential association between these two polymorphisms and EOC. Genotypes were determined for 187 patients with EOC and 221 healthy control subjects, using the MassARRAY system. We found a significant association between the -94 ins/del ATTG genotype distribution and EOC. The frequency of the -94 del ATTG allele was significantly lower in EOC patients compared to healthy controls. The NF-κB mRNA level in cancer tissue was significantly correlated with -94 ins/del ATTG genotypes. Compared to the ATTG1/ATTG1 phenotype, the NF-κB mRNA level was 2.089 and 1.257 times higher in the ATTG2 (insertion)/ATTG2 homozygote and the ATTG1 (deletion)/ATTG2 heterozygote, respectively. However, we found no evidence of association between the 9N ins/del polymorphism of the ß-TrCP gene and EOC in this Chinese population. Based on these results, we suggest that the NF-κB -94 ins/del ATTG polymorphism is a risk factor for EOC susceptibility.

Positive association of major histocompatibility complex class I chain-related gene A polymorphism with leukemia susceptibility in the people of Han nationality of Southern China.

To assess the potential contribution of major histocompatibility complex class I chain-related gene A (MICA) polymorphisms toward the pathogenesis of leukemia, 107 leukemia patients and 162 ethnically matched controls from Hunan province, Southern China, were genotyped for the MICA polymorphism using polymerase chain reaction-sequence-specific priming (PCR-SSP) and sequence-based typing (PCR-SBT). The relevance between these genotypes and risk of leukemia was assessed by means of odds ratio (OR) with 95% confidence intervals (95% CIs). Allele frequencies of MICA-sequence and MICA-STR were different in leukemia patients in comparison with normal controls (both P < 0.05). MICA A5 was directly associated with leukemia (OR = 2.3257, Pc < 0.0005), whereas MICA A5.1 and MICA*008 were inversely associated with leukemia (OR = 0.5874, Pc = 0.0235 and OR = 0.5874, Pc = 0.0329, respectively). In addition, we found that homozygotes for MICA A5 (OR = 14.0659, 95% CI: 3.1627-62.5574, Pc < 0.0001) and MICA*010 (OR = 10.1053, 95% CI: 2.2139-46.1260, Pc < 0.0004) were at an increased risk for leukemia, whereas heterozygotes for MICA*008 and MICA A5.1 were linked to a decreased risk for leukemia (OR = 0.4609, 95% CI: 0.2799-0.7590, Pc = 0.0027). MICA allelic variation is associated with leukemia in Hunan Han population; the data also suggest that MICA gene polymorphism affects susceptibility to different clinical subtypes of leukemia.

Association of major histocompatibility complex class I chain-related gene A microsatellite polymorphism and hepatocellular carcinoma in South China Han population.

The major histocompatibility complex (MHC) class I-related gene A (MICA) is a stress-induced antigen and plays a key role in anti-infectious and anti-tumoral immune responses. In this study, we investigated the MICA transmembrane polymorphism in hepatocellular carcinoma (HCC) patients in South China Han population. We found that the MICA-A5.1 polymorphism, which gives rise to a truncated protein, was increased in HCC patients compared with healthy controls (P(c) = 0.036). Furthermore, we found that HCC patients with MICA-A5.1 polymorphism had higher serum levels of soluble MICA (sMICA). These results suggest that MICA-A5.1 polymorphism is associated with HCC patients in South China Han population and the MICA-A5.1 polymorphism may contribute to the development of HCC by promoting the release of sMICA to evade tumor immunosurveillance.

Purification of lectin from larvae of the fly, Musca domestica, and in vitro anti-tumor activity in MCF-7 cells.

A new lectin was purified from larvae of the fly, Musca domestica L. (Diptera: Muscidae) (MLL-2, 38 kDa) using affinity chromatography and HPLC. Anti-tumor activity of MLL-2 was demonstrated by its inhibition of proliferation of human breast cancer (MCF-7) cells in a time-and dose-dependent manner. The results of acridine orange staining indicated that MLL-2 caused apoptosis in MCF-7 cells. DNA fragmentation in MCF-7 cells has been detected by TUNEL. Flow cytometric analysis also demonstrated that MLL-2 caused dose-dependent apoptosis of MCF-7 cells through cell arrest at G2/M phase. The MLL-2 induced a sustained increase in concentration of intracellular free calcium. Western blot revealed that MLL-2 induced apoptosis in MCF-7 cells was associated with typical apoptosis proteins in the mitochondrial pathway. In addition, the caspase-3 activity in MCF-7 cells treated with MLL-2 for 48 hours was significantly increased compared to controls (407.4 ± 3.0 vs. 1749.2 ± 6.0, P <0.01). Since MLL-2 induced apoptosis in MCF-7cells the mitochondrial pathway may be the main pathway of antitumor activity.

Association study of polymorphisms in the promoter region of DRD4 with schizophrenia, depression, and heroin addiction.

This study investigated the possible association between three functional polymorphisms in the promoter region of the dopamine D4 receptor (DRD4) gene and schizophrenia, depression, and heroin addiction. Genomic DNA was isolated from the venous blood leukocytes of 322 unrelated patients with schizophrenia, 156 patients with depression, 300 patients with heroin addiction, and 300 healthy unrelated individuals. Polymorphisms in the promoter region of DRD4 (-120 bp duplication, -616C/G, and -521C/T) were genotyped using allele-specific polymerase chain reaction analysis. Genotype and allele were analyzed using SPSS 11.5 software. Results of this analysis indicated that there is a strong finding of -120 bp duplication allele frequencies with schizophrenia (p=0.008) and weak finding with -1240 L/S and for paranoid schizophrenia (p=0.022). Interestingly, there is a stronger finding with -521 C/T allele frequencies with heroin dependence (p=0.0002). These observations strongly suggest that the -120-bp duplication polymorphism of DRD4 is associated with schizophrenia and that the -521 C/T polymorphism is associated with heroin addiction.