Psychotropic medication treatment patterns in community-dwelling schizophrenia in China: comparisons between rural and urban areas.

BACKGROUND: To date no study has compared more specifically the psychotropic medication treatment patterns for patients with schizophrenia living in community between rural and urban areas. This study examined the rural-urban differences of the use of psychotropic drugs among community-dwelling individuals with schizophrenia in China. METHOD: Data on 993 community-dwelling patients with schizophrenia (n = 479 in rural area and n = 514 urban area) were collected by interviews during 2013-2014, and 2015-2016 according to the diagnosis of DSM-IV or ICD-10. Data on patients' socio-demographic and clinical characteristics, prescriptions of psychotropic drugs were collected using a standardized protocol and data acquisition procedure. RESULTS: Multivariate analyses revealed that in comparison with the rural counterparts, the patients from the urban area were significantly more frequently prescribed antipsychotic polypharmacy, clozapine, and benzodiazepines, but the patients from the rural area had more frequently prescribed anticholinergics. CONCLUSIONS: Substantial variations in psychotropic medication treatment patterns for patients with schizophrenia living in community were found between rural and urban areas in China. Common use of antipsychotic polypharmacy, clozapine and benzodiazepines in urban area, and anticholinergics in rural area need to be further addressed.

Sexual dysfunction and its impact on quality of life in Chinese patients with schizophrenia treated in primary care.

PURPOSE: Sexual dysfunction in schizophrenia patients is common. In China, maintenance treatment for clinically stable patients with schizophrenia is usually provided by primary care physicians. Illness- or treatment-related sexual dysfunction in this patient population has been never studied. This study describes the prevalence and correlates of sexual dysfunction and its impact on quality of life (QOL) in patients with schizophrenia treated in primary care in China. METHOD: A total of 607 patients with schizophrenia treated in 22 randomly selected primary care services in China formed the study sample. Patients' socio-demographic and clinical characteristics including sexual function and QOL were recorded using a standardized protocol and data collection. RESULTS: Sexual dysfunction was present in 69.9% of all patients; 60.7% in males and 80.6% in females. Multiple logistic regression analysis revealed that female gender, being single, older age and use of first-generation antipsychotics were independently and significantly associated with more sexual dysfunction accounting for 23.5% of its variance (P<0.001). Unexpectedly, sexual dysfunction was not associated with lower QOL. CONCLUSIONS: High rate of sexual dysfunction was reported in the majority of patients with schizophrenia treated in primary care in China. Given its negative impact on social adjustment, QOL and treatment adherence, efforts should be made to address sexual dysfunction in this patient population.

Antipsychotic polypharmacy and quality of life in patients with schizophrenia treated in primary care in China.

OBJECTIVE: In China, maintenance treatment for clinically stable patients with schizophrenia is usually provided by primary care physicians, but their prescribing patterns have not been studied. This study examined the frequency as well as demographic and clinical correlates of antipsychotic polypharmacy (APP) and its impact on quality of life (QOL) in patients with schizophrenia treated in primary care in China. METHOD: A total of 623 community-dwelling patients from 18 randomly selected primary care services were interviewed. Patients' socio-demographic and clinical characteristics, including number of hospitalizations, antipsychotic drug-induced side effects, and QOL were recorded using a standardized protocol and data collection procedure. RESULTS: The rate of APP prescription was 31% (193/623). Of the patients on APP, 89.6% received 2 antipsychotics, 10.4% received 3 or more antipsychotics. Clozapine (35.6%) was the most commonly prescribed second generation antipsychotic (SGA), while perphenazine (17.8%) was the most commonly prescribed first generation antipsychotic (FGA). Multiple logistic regression analyses revealed that patients on APP were more likely to receive SGAs and anticholinergics, had fewer hospitalizations, younger age of onset, and higher doses of antipsychotics. There were no significant differences between the two groups in any of the QOL domains. CONCLUSIONS: Approximately a third of Chinese patients with schizophrenia in primary care receive APP. Further examination of the rationale and appropriateness of APP and its alternatives is warranted.

Comorbid Moderate-Severe Depressive Symptoms and their Association with Quality of Life in Chinese Patients with Schizophrenia Treated in Primary Care.

This study described the prevalence and correlates of comorbid moderate-severe depressive symptoms (comorbid depression thereafter) and their association with quality of life (QOL) in schizophrenia patients treated in primary care. 623 schizophrenia patients were enrolled. Patients' socio-demographic and clinical characteristics including comorbid depression [defined as a total score of 18 or above on the Montgomery-Asberg Depression Rating Scale (MADRS)] were recorded. Depressive symptoms (defined as a total score of 9 or above on the MADRS) were present in 54.1 % of patients, while 17.7 % had comorbid depression. Analysis of covariance revealed that comorbid depression was significantly associated with lower mental QOL. Multiple logistic regression analysis revealed that more severe positive and negative symptoms, anxiety symptoms, use of first-generation antipsychotics and antidepressants, were independently associated with comorbid depression. Given the negative association between comorbid depression and QOL, attempts to address comorbid depression in schizophrenia patients treated in primary care should be made.

Prenatal diagnosis of 15q11.2 microdeletion fetuses in Eastern China: 21 case series and literature review.

OBJECTIVE: 15q11.2 microdeletion can lead to syndromes affecting the nervous system. However, 15q11.2 microdeletion has large phenotypic differences and incomplete penetrance, which brings challenges to prenatal diagnosis. We reported 21 cases of 15q11.2 microdeletion fetuses in Eastern China and reviewed literature on the prenatal clinical characteristics related to the deletion variants to provide a basis for prenatal genetic counseling. METHODS: The clinical data of 21 cases of 15q11.2 microdeletion fetuses collected from June 2018 to September 2021 were retrospectively analyzed, and chromosomal microarray analysis was performed. The reported prenatal clinical features of 15q11.2 microdeletion fetuses were reviewed and summarized. A meta-analysis of 20 studies was performed to test heterogeneity, data integration, and sensitivity on the correlation between 15q11.2 microdeletion and neuropsychiatric diseases. RESULTS: The median age of the women was 29.5 years. The median gestational age at interventional examination was 24 weeks. All fetuses showed deletion variants of the 15q11.2 fragment, and the median deletion range was approximately 0.48 MB. Ultrasound of five cases showed no abnormalities; however, four of them showed a high risk of Down's syndrome (risk values were 1/184, 1/128, 1/47, and 1/54, respectively). The remaining 16 fetuses showed congenital heart disease (7/16), elevated nuchal translucency (5/16), abnormal brain structure (2/16) and renal disease (2/16). In a literature review of 82 prenatal cases, 44% (36/82) had abnormal ultrasound features, 31% (11/36) showed abnormal nuchal translucency, approximately 28% (10/36) showed abnormal cardiac structure, and 14% (5/36) had brain structural abnormalities. The meta-analysis revealed that the frequency of the 15q11.2 microdeletion mutation in patients with schizophrenia and epilepsy was significantly higher (odds ratio 2.04, 95% confidence interval: 1.78-2.33, p < 0.00001; odds ratio 5.23, 95% confidence interval: 2.83-9.67, p < 0.00001) than that in normal individuals. CONCLUSION: More than half of the 15q11.2 microdeletion cases presented no abnormalities in prenatal ultrasound examination. The cases with ultrasound features mainly showed isolated malformations such as elevated nuchal translucency, congenital heart disease, and brain structural abnormalities. Postpartum 15q11.2 microdeletion patients are at an increased risk of suffering from schizophrenia, epilepsy, and other neurological and mental diseases from 15q11.2 microdeletion. Therefore, prenatal diagnosis of 15q11.2 microdeletion not only depends on molecular diagnostic techniques but also requires cautious genetic counseling.

[Mutation analysis of SMN gene in a patient and his family with spinal muscular atrophy].

OBJECTIVE: To perform mutation analysis and describe the genotype of the SMN gene in a patient with spinal muscular atrophy (SMA) and his family. METHODS: Deletion analysis of the SMN1 exon 7 by conventional PCR-restriction fragment length polymorphism (RFLP) and allele-specific PCR, and gene dosage of SMN1 and SMN2 by multiplex ligation-dependent probe amplification (MLPA) were performed for the patient and his parents; reverse transcriptase (RT)-PCR and sequencing were performed for the patient. To determine whether the SMN variant was exclusive to transcripts derived from SMN1, the RT-PCR product of the patient was subcloned and multiple clones were sequenced directly; PCR of SMN exon 5 from the genomic DNA of the parents and direct sequencing were performed to confirm the mutation. RESULTS: In SMN1 exon 7 deletion analysis, no homozygous deletion of the SMN1 was observed in the family; the gene dosage analysis by MLPA showed that the patient had 1 copy of SMN1 and 1 copy of SMN2 his father had 2 copies of SMN1 and 2 copies of SMN2, and his mother had 1 copy of SMN1 and no SMN2. A previously unreported missense mutation of S230L was identified from the patient and this mutation was also found in his father. CONCLUSION: A novel missense mutation of S230L was identified in the SMA family and the genotype of the family members were investigated.

Nicotine dependence in community-dwelling Chinese patients with schizophrenia.

BACKGROUND: Smoking is a serious public health problem. Patients with schizophrenia usually have a higher prevalence of smoking than the general population, but the level of nicotine dependence is seldom studied, especially for patients living in the communities. AIMS: This study aimed to examine the level of nicotine dependence in Chinese community-dwelling patients with schizophrenia and explored its associated sociodemographic and clinical factors. METHODS: A total of 621 patients with schizophrenia treated in the primary care centres of Guangzhou were consecutively recruited. The level of nicotine dependence was assessed with the Chinese version of the Fagerström Test for Nicotine Dependence (FTND). RESULTS: 148 patients with schizophrenia were current smokers, and the mean (SD) score of FTND was 5.06 (2.55) for all the current smokers. The prevalence of nicotine addiction was 48.0% (95% CI: 40.0%-56.0%) in patients with current smoking. The patients with schizophrenia had a significantly higher level of nicotine dependence than the Chinese general population. Multiple linear regression analysis revealed that male gender, being unemployed, having a family history of psychiatric disorders, having major medical conditions, first illness episode and less severe positive symptoms were significantly associated with a higher level of nicotine dependence. CONCLUSION: Community-dwelling patients with schizophrenia in China, especially male patients, had a higher level of nicotine dependence than the general population.

[Molecular diagnosis of spinal muscular atrophy by multiplex ligation-dependent probe amplification].

OBJECTIVE: To investigate the effect of multiplex ligation-dependent probe amplification (MLPA) in molecular diagnosis of spinal muscular atrophy (SMA). METHODS: Peripheral blood samples were collected from 13 SMA patients, 31 parents of SMA patients, 50 healthy individuals without family history of SMA, and 10 specimens of amniotic fluid from these families were collected too. Genomic DNA was analyzed by MLPA, conventional PCR-RFLP, and allele-specific PCR. RESULTS: In complete agreement with the results of conventional PCR-RFLP and allele-specific PCR, MLPA analysis showed that all of the 13 patients had homozygous deletion of the survival of motor neuron 1 (SMN1) gene, and there was significant difference between the SMA severity (type I to type III) and SMN2 copy number (P < 0.05). Of the 31 parents 29 (93.5%) had 1 copy of SMN1, 2 (6.5%) had 2 copies of SMN1. Of the 50 healthy individuals, 1 (2.0%) had 1 copy of SMN1, 48 (96.0%) had 2 copies of SMN1, and 1 (2.0%) had 3 copies. The SMN1 copy number of the parents was significantly higher than that of the healthy individuals (P < 0.01). Two of the 10 fetuses had homozygous deletion of SMN1. CONCLUSION: The MLPA technique has proved to be an accurate and reliable tool for the molecular diagnosis of SMA, both in patients and in healthy carriers.

Antipsychotic-Free Status in Community-Dwelling Patients With Schizophrenia in China: Comparisons Within and Between Rural and Urban Areas.

OBJECTIVE: To date, no study has specifically compared antipsychotic-free patients with schizophrenia living in the community between rural and urban areas. This study examined the rural-urban differences among antipsychotic-free community-dwelling individuals with schizophrenia in China. METHODS: Data on 1,365 community-dwelling patients with schizophrenia (n = 742 in a rural area and n = 623 in an urban area) with diagnoses according to DSM-IV or ICD-10 were collected by interviews during 2013-2014 and 2015-2016. Data on patients' sociodemographic and clinical characteristics, prescriptions of psychotropic drugs, and antipsychotic treatment status were recorded using a standardized protocol and data collection procedure. RESULTS: The prevalence of antipsychotic-free status in the total sample (N = 1,365) was 27.3%; the proportion of antipsychotic-free patients was significantly lower (17.5%) in the urban area (17.5%) than in the rural area (35.4%; χ² = 55.03, P < .001). Binary logistic regression analysis revealed that antipsychotic-free patients, whether from the urban area or the rural area, were older (P = .001, odds ratio [OR] = 0.95 in urban; P = .006, OR = 0.97 in rural) and had poorer attitude toward medication treatment (P < .001, OR = 1.21 in urban; P < .001, OR = 1.31 in rural). Antipsychotic-free patients from the urban area also had fewer admissions, lower education level, and greater likelihood of living by themselves. Antipsychotic-free patients from the rural area also had worse insight into the disease, fewer anxiety symptoms, more prominent positive symptoms, and lower body mass index and were more likely to be women. CONCLUSIONS: Antipsychotic-free status was more common in community-dwelling patients with schizophrenia in the rural area than in the urban area. Older age and poorer attitude toward medication treatment were common features of antipsychotic-free patients. There were correspondingly different risk factors for antipsychotic-free status between rural and urban areas. Building a positive medication treatment attitude is an important strategy for establishing medication adherence in older, community-dwelling patients with schizophrenia.

Quality of life in Chinese patients with schizophrenia treated in primary care.

In China, maintenance treatment for clinically stable patients with schizophrenia is usually provided by primary care physicians. This study examined the quality of life (QOL) in patients with schizophrenia treated in primary care and explored the demographic and clinical characteristics associated with QOL. Altogether, 612 patients with schizophrenia treated in 22 randomly selected primary care services in China formed the study sample. QOL, psychotic and depressive symptoms, extra-pyramidal symptoms and insight were assessed using standardized instruments. Data analyses were conducted with the one sample t-test and multiple linear regression analyses. Compared with the normative data for the Chinese general population, significantly lower scores in physical and mental QOL domains were found in the patient group. Older age, being unemployed, major medical conditions, no smoking, more severe depressive and negative symptoms, more frequent insomnia, and suicidality were independently associated with poor physical QOL. Male gender, more severe depressive and anxiety symptoms, more frequent insomnia, and suicidality were independently associated with poor mental QOL. Patients with schizophrenia treated in primary care had lower level of QOL in comparison with general population. Effective measures need to be implemented to improve their QOL.

Prevalence of Insomnia and Clinical and Quality of Life Correlates in Chinese Patients With Schizophrenia Treated in Primary Care.

PURPOSE: To describe the prevalence and clinical correlates of insomnia in schizophrenia patients treated in primary care. DESIGN AND METHODS: Six hundred and twenty-three schizophrenia patients from 22 primary care services were recruited. FINDINGS: The prevalence of at least one type of insomnia was 28.9% (180/623), while those of difficulty initiating sleep, difficulty maintaining sleep, and early morning wakening were 20.5%, 19.6%, and 17.7%, respectively. Only 53.3% of patients suffering from insomnia received treatment. PRACTICE IMPLICATIONS: Insomnia is common in Chinese patients with schizophrenia treated in primary care and the rate of treatment appears low.

The Relationship Between Sleep Patterns, Quality of Life, and Social and Clinical Characteristics in Chinese Patients With Schizophrenia.

PURPOSE: The purpose of the study was to determine the pattern of sleep behavior in schizophrenia patients treated in primary care. DESIGN AND METHODS: Altogether 623 schizophrenia patients in 22 primary care services were recruited. Sleep duration and demographic and clinical characteristics were recorded. FINDINGS: The mean expected total sleep time was 8.8 hr (SD 1.8) and the mean actual total sleep time was 8.2 hr (SD 2.1). The frequency of short, medium, and long sleepers was 18.1, 38.4, and 43.5%, respectively. Major medical conditions and any type of insomnia were independently associated with short sleep, while long sleep was associated with unemployment and use of second-generation antipsychotics. PRACTICE IMPLICATIONS: More attention should be paid to sleep duration in this population group.

[Study of fetal lymphocyte of intrahepatic cholestasis of pregnancy].

OBJECTIVE: To explore effect of fetal lymphocyte on pathogenesis of intrahepatic cholestasis of pregnancy (ICP). METHODS: Twenty pregnant women with ICP and 20 normal pregnant women were enrolled in the study. The single mixed lymphocyte culture/reaction (MLC/MLR) was conducted using inactive lymphocyte obtained from maternal peripheral blood and lymphocyte of cord blood from fetus. Antigen-induced-lymphocyte-proliferation-reaction was used for dermic soluble antigen and decidual soluble antigen obtained from maternal blood and cord blood from fetus. The intense of proliferation was calculated and compared between normal and ICP-complicated pregnancies. RESULTS: (1) The level of intense of proliferation of fetal lymphocyte was significantly increased in ICP group 2.75 +/- 0.36 than those of normal control group 1.45 +/- 0.19 in single mixed lymphocyte culture (P < 0.05). (2) The level of intense of proliferation of fetal lymphocyte was significantly increased in ICP group 1.45 +/- 0.19 than those of normal control group 0.67 +/- 0.24 in decidual soluble antigen induced lymphocyte proliferation reaction (P < 0.05). (3) The level of intense of proliferation of fetal lymphocyte was significantly increased in ICP group (1.22 +/- 0.44) than those of normal control group (0.66 +/- 0.27) in dermic soluble antigen induced lymphocyte proliferation reaction. CONCLUSIONS: (1) The fetal lymphocyte may be one of the effector cells in pathogenesis of ICP. (2) The disturbance of fatal-maternal immune-tolerance is one of the important mechanisms underlying ICP.

[Cytotoxicity of cytotoxic T lymphocytes induced by the dendritic cells phagocytosing PLA-AFP218-226 micospheres Against hepatocellular carcinoma cell lines].

OBJECTIVE: To assess the cytotoxicity of cytotoxic T lymphocytes (CTLs) induced by the dendritic cells phagocytosing HLA-A2+ restricted epitope peptides encapsulated in polylactic acid (PLA) microspheres (PLA-AFP218-226) against cell lines HepG2 and T2-loaded with HLA-A2+ restricted epitope peptides derived from alpha fetoprotein (AFP218-226, LLNQHACAV). METHODS: Mature dendritic cells (DCs) were obtained by inducing the monocytes isolated from peripheral blood cells of HLA-A2+ healthy donors with GM-CSF and IL-4. On day 3 from onset of the culture, PLA- AFP218-226 was added to the culture medium, and on day 6, lipoplysaccharide (LPS) was added to it for inducing the immature DCs to mature. The surface phenotype of the mature DCs was determined with fluorescence activated cell sorting (FACS) assay; the cytotoxicity of CTLs induced by the DCs for 7 days against cell lines HepG2 and T2-loaded with AFP218-226 was determined with MTT method; and the avidity between HLA-A2 and AFP218-226 was determined with T2-peptide binding experiment. RESULTS: There was high avidity between AFP218-266 and HLA-A2. The DCs phagocytosing PLA-AFP218-226 highly expressed CD83, CD86, CD40, etc., and the CTLs induced by the DCs strongly decomposed the HepG2 and T2-loaded with AFP218-226. CONCLUSIONS: The strong cytotoxicity against HepG2 cell lines can be induced in vitro by DCs phagocytosing PLA-AFPM218-226 micospheres, suggesting that PLA-AFP218-226 microspheres can serve as a new type of CTL epitope vaccine for the prophylaxis and treatment of hepatocellular carcinoma.

[Gene Diagnosis and Analysis of Clinical Hematological Phenotype of Thailand Deleted α-Thalassemia 1].

OBJECTIVE: To investigate the hematologic characteristics and gene diagnosis of patients with Thailand deleted α-thalassemia 1, so as to provide the information for clinical genetic counseling. METHODS: The clinical data of 32 patients with Thailand delated α-thalassemia 1 were analyzed retrospectively; the hematologic characteristics and gene diagnosis of Thailand deleted type were investigated by using routine hematologic examination, genetic detection of common thalassemia and Thailand deleted α-thalassemia 1. RESULTS: Among 32 cases, the Thailand deleted α-thalassemia 1 heterozygote was found in 29 cases, the Thailand deleted α-thalassemia 1 and α(3.7) gene deletion double heterozygote were found in 1 case, the Thailand deleted α-thalassemia 1 with ß-thalassemia (1 case with codons 41-42 mutation heterozygous, 1 case with CD17 mutation heterozygous) was found in 2 cases by detection. The MCV and MCH levels were decreased in all cases of Thailand deleted thalassemia 1, there were significant differences in RBC, MCV, MCH (P<0.05) between normal control and Thailand deletion α-thalassemia 1 group; there were also significant differences in MCHC (P<0.05) between Southeast asia thalassemia and Thailand deleted α-thalassemia 1 group. CONCLUSION: There are no significant differences in hematological parameters except MCHC between Southeast asia thalassemia and Thailand deleted α-thalassemia 1 group. moreover the Thailand deleted α-thalassemia 1 in a certain proportion exists in area with high incidence of thalassemia, therefor the clinicians should pay more attention to the screen and diagnosis of Thailand delated α-thalassemia and can exactly diagnose the Thailand delected α-thalassemia 1 on the basis of comprehensive analysis of conventional and Thailand delected α-thalassemia 1 detection results, clinical presentation, hematologic parameters and ultrasonic examination, so as to avoid the birth of child with severe and intermidiate type α-thalassemia caused by Thailand deleted α-thalassemia 1.

Smoking and its associations with sociodemographic and clinical characteristics and quality of life in patients with schizophrenia treated in primary care in China.

OBJECTIVE: Maintenance treatment for clinically stable patients with schizophrenia is usually provided by Chinese primary care physicians, but no study has investigated smoking rates in this population. This study investigated the rate of smoking and its associations with sociodemographic and clinical characteristics and quality of life (QOL) in patients with schizophrenia treated in primary care in China. METHODS: This was a cross-sectional, community-based survey. A total of 621 schizophrenia patients were recruited from 22 primary care services in Guangzhou, China, in 2013. Patients' sociodemographic and clinical characteristics, smoking status, and QOL were recorded. RESULTS: The frequency of current smoking was 23.8% in the whole sample; 41.5% for men and 2.5% for women. Multiple logistic regression analysis revealed that male gender, married status, alcohol use, older age at onset, fewer major medical conditions, lower education level and more hospitalizations were independently associated with current smoking. CONCLUSION: The frequency of smoking in Chinese schizophrenia patients treated by primary care physicians is lower than most figures reported from Western and Chinese psychiatric settings.

Pivotal molecules of MHC I pathway in human primary hepatocellular carcinoma.

AIM: To investigate the expression of several important molecules involved in major histocompatibility complex (MHC) class I presentation pathway in primary hepatocellular carcinoma (HCC), and to determine whether cytotoxic T lymphocyte (CTL) vaccine therapy was suitable for HCC. METHODS: Labeled streptavidin biotin (LSAB) method of immunohistochemistry was used to study 33 HCC tissue specimens. RESULTS: Most HCC tissues and adjacent histological normal hepatocytes expressed HLA-I antigens,TAP, and B7, expression of B7 was especially strong, and there was no significant difference between them (P>0.05). CONCLUSION: The MHC class I presentation pathway in primary hepatocellular carcinoma may not be abnormal or dysfunctional, and CTL could kill these tumor cells. Thus, it is suitable and practicable to design and construct CTL vaccine against HCC.

[Expression of gastrin in human gastric cancer cell line and effect of anti-gastrin mAb on the cell growth].

OBJECTIVE: To investigate the expression of gastrin in human gastric cancer cell line SGC-7901 and the effects of gastrin-17 and anti-gastrin mAb on its growth. METHODS: The expression of gastrin was determined by immunohistochemistry with anti-gastrin mAb prepared by our group. In a series of experiments, the growth of SGC-7901 cells was evaluated by MTT assay on cells grown in serum-free medium and treated with gastrin-17 and/or anti-gastrin mAb. RESULTS: Immunohistochemical examination of SGC-7901 cells revealed a specific gastrin immunoreactivity. Gastrin-17 significantly stimulated cell growth at the concentrations of 1 x 10(-9) mol/L approximately 1 x 10(-5) mol/L in a dose-dependent manner. The growth of SGC-7901 cells treated with anti-gastrin mAb, either alone or in combination with gastrin-17 (1 x 10(-7) mol/L), was significantly inhibited. CONCLUSION: Growth of human gastric cancer cells SGC-7901 can be stimulated in an autocrine fashion. The gastrin-stimulated growth of gastric cancer cells can be blocked by anti-gastrin mAb bound specifically with gastrin. Further study on the significance of anti-gastrin mAb in designing immunotherapy targeting to gastrin or gastrin receptor is warranted.

[On mechanism of how lipopolysaccharide induces dendritic cells to mature through TLRs signal pathway].

OBJECTIVE: To find out how lipopolysaccharide (LPS) induces dendritic cells (DC) to be more mature through Toll-like receptors (TLRs) signal pathway. METHODS: Monocytes were isolated from heparinized whole blood. 7 days later the monocytes were induced into dendritic cells by adding rhGM-CSF and rhIL-4. Total cellular RNA of unstimulated and stimulated DC were extracted by LPS 24 h later. Semi-quantitative RT-PCR and agarose gel electrophoresis were conducted to analyze the PCR products. The expression of NF-kappaB was detected by laser scanning confocal microscopy (LSCM). RESULTS: Expressions of TLR2, 3, 4 were significantly higher in monocytes-derived DC induced for 7 days, the plasm of DC was stained positive and the nucleus of DC was stained negative. After being stimulated by LPS for 24 h, the expressions of TLR2, 3, 4 decreased, and that of TLR4 was almost undetectable; the nucleus of DC was stained strong positive, the plasm of DC was stained positive. CONCLUSION: The expressions of TLR2, 3, 4 on DC change significantly after being stimulated by LPS. And NF-kappaB shifts from the plasm to the nucleus. This signifies that LPS induces DC to be more mature in function.

[Construction and expression of anti-human AFP ScFv gene in BL-21 (DE3) E. coli].

OBJECTIVE: To construct anti-human AFP single chain fragment variable (ScFv) gene, transform it into BL-21 (DE3) E. coli for expression, and identify its bioactivity. METHODS: VH and VL genes of anti-human AFP monoclonal antibody were cloned by RT-PCR from hybridoma. The ScFv gene was spliced by sequence overlap extending (SOE) PCR, and then it was ligated into pGEM-T vector to be identified by endonuclease digestion, PCR and sequencing. ScFv gene was cloned into pET32 (a+) vector and transformed into BL-21 (DE3) E. coli. The positive clones were screened out by IPTG induction, and the ScFv antibody was purified to be identified by SDS-PAGE and competitive inhibition ELISA test. RESULTS: The VH DNA consisted of 339 bases, coming from the mouse IgG gamma chain. The VL DNA consisted of 312 bases, coming from the mouse IgG kappa chain. The VH and VL genes were spliced by 45 bases coding a (G4S)3 flexible linker. The ScFv gene consisted of 696 bases. The ScFv antibody expressed by BL-21 (DE3) fused with TrxA tag protein and formed inclusions. The relative molecular mass of TrxA-ScFv fusion protein is about 40 x 10(3) and that of ScFv is about 24 x 10(3). The ScFv antibody has excellent activity tested by competitive inhibition ELISA, the TrxA-ScFv could inhibit about 41% of the McAb to bind antigen and ScFv could inhibit about 53%. CONCLUSION: We have successfully constructed an anti-human AFP ScFv gene with 696 bases; it can express in BL-21 with high activity.