Expression and clinical significance of serum ubiquitin in esophageal squamous cell carcinoma / 医学研究生学报

Objective　Ubiquitin-proteasome system (UPS) plays a central role in the development of esophageal cancer. However, As the executor of UPS, the expression and clinical significance of detection of serum ubiquitin (UB) in Esophageal Squamous Cell Carcinoma(ESCC) patients have not been fully elucidated. The aim of this study was to investigate the expression and diagnostic value of serum ubiquitin (UB) in ESCC.　Methods　A total of eighty-eight ESCC patients and forty healthy controls from February 2018 to May 2019 at the Affiliated Jiangyin Hospital of Nantong University were enrolled. Serum UB was measured by ELISA, and serum squamous cell carcinoma antigen (SCC) and carcinoembryonic antigen (CEA) were determined by chemiluminescence method. The ROC curve was used to analyze the diagnostic value of each indicator. Besides, the correlation between serum UB and clinicopathological features of ESCC were analyzed.　Results　The level of serum UB in ESCC group was significantly higher than that in the control group[(41.96±3.273)ng/ml vs (80.86±7.993)ng/ml, P<0.05]. The level of serum UB in ESCC patients was related to lymphatic metastasis and tumor stage (P<0.05). The sensitivity of UB, SCC, CEA and the three combined diagnosis of ESCC were 65.9%, 52.3%, 51.1%, and 76.1%, respectively. The AUC under the ROC curve were 0.690, 0.677, 0.635, and 0.795, respectively.　Conclusion　Serum UB is highly expressed in ESCC and is closely related to tumor progression. Combined with SCC and CEA, UB can improve the sensitivity of diagnosis of ESCC and can be used as an effective serological screening biomarker.

Preliminary research on prokaryotic expression and immune protection of triosephosphate isomerase of Toxoplasma gondii / 中国血吸虫病防治杂志

Objective To study the prokaryotic expression and immune protection of triosephosphate isomerase(TPI)of Toxoplasma gondii in mice. Methods Total RNA was extracted from toxoplasma tachyzoites,and TPI fragment was amplified by PCR and cloned into the prokaryotic expression vector pET-28a(+). The target protein was induced with IPTG and purified by Ni-NTA affinity chromatography. The mice were immunized 4 times by emulsified TPI with adjuvant,and the last time was the strengthen immunization. At the same time,an adjuvant group and a normal group were set as controls. The blood samples were got from the tail vein of the mice,and the serum antibody titres were detected. All the mice were challenged with 400 toxo-plasma tachyzoites to observe the survival time. Results The TPI gene was amplified from T. gondii cDNA by PCR. The recom-binant vector TPI/pET-28a(+)was usefully constructed,and the TPI protein was expressed and purified. The serum antibody ti-tre could be more than 100 thousand. After infected with toxoplasma tachyzoites,the survival time of the mice in the experimen-tal group was longer than that of the mice in the control groups. Conclusion The TPI protein of T. gondii could trigger the im-munoprotection against T. gondii challenge in the mice.

Impact of one-child on body mass index and percentage of body fat in primary school students: a longitudinal study / 上海交通大学学报（医学版）

Objective·To evaluate the impact of one-child on body mass index (BMI), and percentage of body fat in primary school students. Methods·All the sample was recruited from 5 elementary schools in Gaohang Town, Shanghai, China. The baseline data of height, body weight, and percentage of body fat was obtained in 2013, and re-measured in 2014 and 2015, respectively. Birth weight, breast feeding, diet and time for physical activities of each children and the highest education level, height, and body weight of their parents were also collected by a self-completed questionnaire. Logistic regression and Mix model was used to analyze the relationship between one-child and BMI, BMI-Z score, and percentage of body fat. Results·A total number of 2515 (1323 boys and 1192 girls) primary school students completed the study and entered the analysis. The percent of one child in this study population was 72.0％ (1812/2515). BMI-Z score, time for physical activities, parental education level, and maternal BMI were higher, while the rate of breast feeding was lower in one-child group compared to non-one-child group. The results of Logistic regression showed boys (compared to girls), macrosomia ( ≥4000g vs normal birth weight), overweight father and mother (compared to normal BMI) were risk factors for overweight. The factor of one-child didn't increase the risk of overweight (OR=1.119, 95％ CI 0.911-1.374). After potential con-founders adjusted, the annual increase of BMI (β=0.028, 95％ CI -0.045-0.100), BMI-Z score (β=0.002, 95％CI -0.034-0.037) and percentage of body fat (β=0.013, 95％ CI -0.181-0.207) showed no obvious difference between the two groups. Conclusion·One-child factor showed no obvious relationship with BMI, BMI-Z score and percentage of body fat in primary school students.

Construction and evaluation of charge-reversal nanocarrier loaded with hematoporphyrin monomethyl ether / 药学学报

Charge-reversal nanocarrier was constructed to enhance lysosomal escape and improve an-titumor effect. We synthesized the cholesterol-polyethyleneimine-hexahydrophthalic anhydride (Chol-PEI-HHPA) polymer and characterized by 1H NMR. The charge-reversal liposomes (Lipo-HHPA) were synthesized and the hematoporphyrin monomethyl ether (HMME) was loaded. pH-triggered charge conversion was determined at different pH values. The lysosomal escape and cytotoxicity of the Lipo-HHPA were evaluated in MCF-7 cells. The Lipo-HHPA was uniform with an average particle size of 102 nm. Upon the irradiation of ultrasound, burst release of HMME could be observed. The zeta potential of Lipo-HHPA changed sharply from negative (-23.5 mV) to positive (+21.2 mV) over the pH range of 7.4-4.5. In the cellular uptake experiment, the lysosomal escape of Lipo-HHPA was observed. HMME loaded Lipo-HHPA displayed obviously enhanced cytotoxicity towards MCF-7 cells. These results indicate that the charge-reversal liposomes hold a great potential in improving the cytotoxicity and antitumor effect.

Expression level of microRNA-92a and its clinical significance in multiple myeloma patients / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the relationship between the expression level of microRNA 92a (miR-92a) and del(13q14) and the prognosis of MM patients, and to explore the pathway that miR-92a involved.</p><p><b>METHODS</b>Bone marrow samples from 53 newly diagnosed MM patients were collected, del(13q14) was analyzed by interphase fluorescence in situ hybridization in sorted CD138 positive plasma cell. The expression of miR-92a in plasma cells was measured by quantitative real-time PCR. The expression of c-jun was detected by Western blot in miR-92a transfected MM cell lines (LP-1, U266 and JJN3).</p><p><b>RESULTS</b>Of the 53 MM patients, del(13q14) was detected in 31 (58.4%) patients. The median levels of miR-92a in MM patients with or without del(13q14) were 27.36±2.61 and 21.87±15.98, respectively (P>0.05). With the median follow-up of 13.5 (0.5-72.5) months, the median duration of progression-free survival of patients with high expression level of miR-92a was significantly shorter than those with low expression level of miR-92a (4.5 months vs 14.0 months, P=0.006). Overexpression of miR-92a in MM cell lines induces time-dependent down-regulation of c-jun.</p><p><b>CONCLUSIONS</b>High expression of miR-92a was associated with poor prognosis in MM patients. The expression level of miR-92a was not associated with del(13q14), and the effect of miR-92a on the progress of MM might be involved in c-jun pathway.</p>

Effects of comprehensive therapy on serum SPARC levels in ankylosing spondylitis patients accompanied with osteoporosis / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the effects of comprehensive therapy on serum secreted protein acidic and rich in cysteine (SPARC) levels in ankylosing spondylitis (AS) patients accompanied with osteoporosis (OP), and to explore the possible mechanisms for SPARC in AS patients accompanied with osteoporosis.</p><p><b>METHODS</b>Totally 48 AS patients accompanied with OP (Group A) were treated with massage, intravenous infusion of Cervus and Cucumis Polypeptide Injection, and Bushen Quhan Zhiwang Decoction (BQZD) for 3 months. At the same time, 45 normal healthy subjects were recruited as the normal control group (Group B). Serum SPARC levels were measured by ELISA in Group A before and after comprehensive therapy and in those of Group B. The levels of bone mineral density of femoral neck (FN BMD), bone mineral density of 2 -4 lumbar spine (L2-4 BMD), bone specific alkaline phosphatase (BSAP), tumor necrosis factor alpha (TNF-alpha), and transforming growth factor beta-1 (TGF-beta1) were detected. Meanwhile, Bath AS disease activity index (BASDAI) and Bath AS functional index (BASFI) were detected in Group A before and after treatment. The correlations between the aforesaid indices and serum SPARC levels were analyzed.</p><p><b>RESULTS</b>Serum SPARC levels were significantly lower in those of Group A than in those of Group B (175. 30 +/- 72.04 micro/L vs 190. 52 +/- 86. 13 microg/ L, P <0. 01). Serum SPARC levels in those of Group A were negatively correlated with TNF-alpha (r = -0.261, P <0.01), positively with L2-4 BMD, TGF-beta1, and BSAP (r =0.437,0.256, 0.385, P <0.05, P <0.01). L2-4BMD and BSAP were independently predictors of serum SPARC in patients of Group A. After comprehensive therapy, the levels of TNF-alpha, BASDAI, and BASFI obviously decreased, TGF-beta1, BSAP, L2-4 BMD, and FN BMD obviously increased (P <0. 05, P <0. 01). The serum SPARC levels also significantly increased (188.32 +/- 87.50 microg/L, P <0. 05).</p><p><b>CONCLUSION</b>Comprehensive therapy could effectively improve the bone metabolism, clinical symptoms and the activity function of joints, and elevate serum SPARC levels.</p>

Clinical significance of serum free light chain in patients with multiple myeloma / 中国实验血液学杂志

Multiple myeloma (MM) is a malignant disorder characterized by the proliferation of a single clone of plasma cells that can produce excessive amounts of serum free light chain (sFLC). sFLC plays an important role in MM diagnosis and disease monitoring. The quantitative immuno-nephelometric assay is sensitive and specific means for sFLC testing. The aim of this study was to investigate the levels of sFLC in multiple myeloma and the relationship between sFLC and serum total light chain (sTLC). sFLC in 45 newly diagnosed patients were detected by immuno-nephelometric assay, and then the ratio of free kappa to free lambda for every sample was calculated. Meanwhile, sTLC was also determined in these patients. The results showed that the difference of sFLC levels between MM patients and the normal controls was significant (tΚ = 8.86, P < 0.001; tλ = 15.48, P < 0.001;tΚ/λ = 5.54,P < 0.005). No correlation between sFLC and sTLC was found in MM patients. It is concluded that the level of sFLC in MM patients is significantly higher than that in normal controls. sFLC and its ratio may be served as a indicator for diagnosis of MM. sTLC can not replace the role of sFLC.

Investigation of 1q21 amplification in patients with multiple myeloma using I-FISH and cIg-FISH / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the prevlance of 1q21 amplification in patients with multiple myeloma (MM) and its correlation with the progression and prognosis of the disease.</p><p><b>METHODS</b>1q21 amplification was detected in 48 patients with MM using cytoplasmic light chain immunofluorescence with fluorescence in situ hybridization analysis (cIg-FISH) and interphase fluorescence in situ hybridization (I-FISH) analysis combined with CD138 immunomagnetic cell sorting (MACS).</p><p><b>RESULTS</b>1q21 amplification (≥ 3 red signals) was detected in 26/48(54.2%) cases by cIg-FISH and 31/48 (64.6%) cases by I-FISH combined with CD138 MACS. There was a good consistency between the two methods (P>0.05). The mortality of patients with 1q21 amplification was significantly higher than those without (P< 0.05). No significant difference was detected in terms of sex, age, Durie-Salmon stage, subgroup and international staging system (ISS) stage between patients with 1q21 amplification and those without (P>0.05).</p><p><b>CONCLUSION</b>The frequency of 1q21 amplification in MM is high. There was also an association between the amplification and poor prognosis. cIg-FISH is consistent with CD138 MACS combined with I-FISH.</p>

Role of bone marrow microenvironment in regulation of AP-1 gene expression in multiple myeloma cells / 中国实验血液学杂志

This study was aimed to investigate the role of bone marrow microenvironment in the regulation of activator protein 1 (AP-1) expression in multiple myeloma (MM) cells. The primary myeloma cells (CD138(+) cells) from 8 patients with MM were sorted by using immunomagnetic beads and were cocultured with osteoclasts in alpha-MEM supplemented with 10% fetal bovine serum, antibiotics, RNAKL (50 ng/ml) and macrophage colony-stimulating factor (25 ng/ml) for 10 to 14 days at 2.5 x 10(6) cells/ml. The expression levels of genes c-jun, junD fos and fosB were detected by real-time PCR. The results showed that the osteoclasts were observed after coculture of mononuclear cells in peripheral blood of MM patients with osteoclasts for 10 - 14 days. As compared with control (without coculture with osteoclasts), the viability of MM cells cocultured with osteoclasts obviously increased, the expression levels of c-jun, junD, fos and fosB decreased to 25.7% - 1.66%, 68.49% - 8.54%, 10.35% - 0.19% and 36.63% - 3.44% of the control respectively. It is concluded that the bone marrow microenvironment can inhibit the expression of c-jun, junD, fos and fosB promote myeloma cell proliferation and maintain cell survival.

Investigation of chromosome 1 aberrations in patients with multiple myeloma using cIg-FISH method and its significance / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the incidence of 1q21 amplification and 1p12 deletion, and analyze the correlation between these aberrations with disease progression, prognosis and outcome in patients with multiple myeloma (MM).</p><p><b>METHODS</b>Cytoplasm light chain immunofluorescence with simultaneous interphase fluorescence in situ hybridization (cIg-FISH) was used to detecte the 1q21 amplification and 1p12 deletion in 48 patients with MM.</p><p><b>RESULTS</b>1q21 amplification (≥ 3 red signals) was determined in 26 of 48(54.2%) cases. The mortality of patients with 1q21 amplification was significantly higher than that of those lacking 1q21 amplification (P < 0.05). The sex, age, D-S stage, subgroup and ISS stage between patients with and without 1q21 amplification had no significant difference (P > 0.05). There was a significant difference in D-S stage and mortality between patients with 3 and with 4 copies of 1q21 (P < 0.05). No significant difference in sex, age, subgroup, ISS stage, and isotype was found between them (P > 0.05). 1p12 deletion (< 2 green signals) was found in 14 of 48 (29.2%) cases. There was no significant difference in sex, age, D-S stage, ISS stage, isotype, subgroup, and mortality between patients with and without 1p12 deletion.</p><p><b>CONCLUSION</b>The frequency of chromosome 1 aberrations in multiple myeloma is high and 1q21 amplification is a poor prognosis factor.</p>

Detection of complex chromosomal aberrations in patients with multiple myeloma using multiplex fluorescence in situ hybridization / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To explore the value of multiplex fluorescence in situ hybridization (M-FISH) in the detection of the complex chromosomal aberrations (CCAs) in multiple myeloma (MM).</p><p><b>METHODS</b>M-FISH was used in 10 MM patients with CCAs detected by conventional cytogenetics (CC) using R-banding to refine the rearrangement of CCAs and identify the characteristics of marker chromosome.</p><p><b>RESULTS</b>M-FISH confirmed the 29 structural aberrations shown by CC analysis, and also confirmed the specific source of 21 types of chromosomal aberration, which were not detected by CC analysis. Among them, t(2;15)(q33;q22), t(6;7)(q23;q34), t(8;11) (q24;q23), t(1;14)(q10;q32) and t(X;1)(q26;q25) were new chromosomal aberrations. The median survival time of 9 MM patients with CCAs was 23 months and evidently shorter than that of MM patients without CCAs, with the mean survival time being 34 months.</p><p><b>CONCLUSION</b>M-FISH could refine CCAs in MM patients, find or correct the missed or misidentified abnormalities analyzed by CC. It has provided one of the essential methods for the research of chromosomal aberrations in MM.</p>

Investigation of the molecular changes in patients with multiple myeloma by fluorescence in situ hybridization / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the incidence and prognosis of 1q21 amplification, 13q14 deletion, TP53 gene deletion and IgH translocation in patients with multiple myeloma (MM).</p><p><b>METHODS</b>Interphase fluorescence in situ hybridization (I-FISH) with four different specific probes for the regions containing 1q21, 13q14.3 (D13S319), 14q32 and TP53 gene were performed in 43 MM patients.</p><p><b>RESULTS</b>Among the 43 MM patients, 1q21 amplification was observed in 28 (65.1%) cases, 13q14 deletion in 30 (69.7%) cases, TP53 gene deletion in 8 (18.6%) cases, and IgH translocation in 29 (67.4%) cases. The mortality of MM patients with 1q21 amplification, 13q14 deletion or TP53 gene deletion was higher than those without them.</p><p><b>CONCLUSION</b>There is high frequency of 1q21 amplification, 13q14 deletion, TP53 gene deletion and IgH translocation in multiple myeloma, and 1q21 amplification, 13q14 deletion and TP53 gene deletion are poor prognosis factors for MM patients.</p>

microRNA-21 and microRNA-30b expression in multiple myeloma / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the expression of miR-21 and miR-30b in multiple myeloma (MM).</p><p><b>METHODS</b>Peripheral blood mononuclear cells from patients with MM were cultured at 2.5 x 10(6) cells/ml in alpha-MEM supplemented with 10% of fetal bovine serum, antibiotics, RANKL (50 ng/ml), and macrophage colony-stimulating factor (25 ng/ml) for 10 to 14 days to obtain osteoclasts with bone-resorbing activity. Primary myeloma cells were purified from 12 MM patients. Of them, 8 samples were cocultured with osteoclasts and 4 as noncocultured control. The expression of miR-21 and miR-30b was detected by real-time PCR.</p><p><b>RESULTS</b>The viability of MM cells recovered from cocultures was higher than those of noncocultured control. After cocultured with osteoclasts, primary myeloma cells from eight patients exhibited a 1.3- 5.9-fold increase in miR-21 expression and 1.38- 4.32-fold decrease in miR-30b expression compared with controls. In highly purified plasma cells from 3 healthy subjects, 12 MM patients and 11 MM cell lines, the expression of miR-21 was 1.9 +/- 0.8, 6.5 +/- 4.9 and 35.1 +/- 36.2, respectively; the expression of miR-30b was 13.6 +/- 1.8, 7.2 +/- 6.3 and 4.5 +/- 1.9, respectively.</p><p><b>CONCLUSIONS</b>miR-21 acts as an oncogene and miR-30b a tumor suppressor gene in MM.</p>

The preparation of quality control materials for human papillomavirus genotyping / 中华检验医学杂志

Objective To prepare the HPV genotyping control materials. Methods Three hundred cervical smears with different HPV genotypes were collected and detected by surface plasmon resonance and sequencing. The primers for specific genotype were designed according to GenBank. The recombinant plasmid was constructed through PCR amplification, ligation and transformation. Thirty recombinant plasmids were identified through PCR amplification, sequencing, and the sequences were compared using BLAST. Results The collected HPV infectious samples contained 30 different genotypes including HPV 6,16,18 and so on.The fragment sequences of PCR amplification were concordant with the designed. The fragment sizes of the other types ranged from 1 500 to 2 000 bp except HPV CP8304. And the 30 recombinant plasmids identified by PCR were concordant with the target. The identity of BLAST was 99%. In the fragment of 1500 bp in length, 11 bases were inconsistent with the reference sequence. Conclusions Genotyping control materials were developed successfully. The human papillomavirns genotyping control materials covered all the most common types and included 14 types with high-risk, 3 types with medium-risk and 13 types with low-risk.

Fluorescence in situ hybridization on bone marrow smear in the detection of cytogenetic aberrations of multiple myeloma / 中国实验血液学杂志

This study was aimed to establish the technique of interphase fluorescence in situ hybridization (I-FISH) used on smear of bone marrow directly, and to develop a new method for detection of the molecular cytogenetics in multiple myeloma (MM). After a series of treatment, fixation and digestion of the bone marrow smear as the carrier, the chromosome 8 centromere probe were used in I-FISH for molecular cytogenetics detection. At the same time, differences were compared in the results between the new method and the conventional I-FISH. The results showed that there was no statistically significant difference of proportion of various signals in non-hematologic malignancies when detected with the two methods (p>0.05). In bone marrow smear I-FISH, 8 out of 19 cases (42.1%) had abnormality of chromosome 8, including 5 cases with -8 (26.3%) and 3 cases with +8 (15.8%). It is concluded that the I-FISH on smear of bone marrow is characterized by convenience, economy and accuracy. Therefore, it can be used for research of molecular cytogenetics in MM.

Correlation between chromosome 13q14 deletion and 1q abnormality in multiple myeloma / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the correlation between chromosome 13q14 deletion [del(13q14)] and chromosome 1q abnormality in multiple myeloma (MM).</p><p><b>METHODS</b>The bone marrow plasma cells of 48 previously untreated MM patients were purified by CD138 and magnetic cell sorting system, and interphase fluorescence in situ hybridization (I-FISH) was applied to detect the del(13q14) with D13S319 probe and the abnormalities of chromosome 1q with CEP1 SpectrumOrange probe in sorted MM cells.</p><p><b>RESULTS</b>Among the 48 MM patients, del(13q14) was observed in 22(45.8%) cases, the abnormalities of chromosome 1q were observed in 23 (47.9%) cases, among which 2 were 1q deletion and 21 were 1q duplication. The chromosome 1q abnormality was detected in 16 of the 22 cases of MM with del(13q14) and in 7 of the 26 cases of MM without del(13q14), and there was significant difference between the two groups (chi-square was 10.02, P was less than 0.01).</p><p><b>CONCLUSION</b>There is high frequency of chromosome 13q14 deletion and 1q abnormality in multiple myeloma. The chromosome 1q abnormalities are highly associated with 13q14 deletion.</p>

Reviews on antiviral activity of chemical constituents from plants / 中国中药杂志

This paper reviewed the progress in researches on antiviral activity of chemical constituents from plants in recent years, the antiviral activity and mechanism of action of flavonoids, alkaloids, terpenoids, coumarins and polysaccharoses were sammarszed, provided new leading compound for antivirus new drugs from the plares in prospect.

Effects of bushen tongdu decoction on serum tumor necrosis factor-alpha and transforming growth factor beta1, in patients with ankylosing spondylitis / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the relationship of tumor necrosis factor-alpha (TNF-alpha) and transforming growth factor beta1 (TGF-beta1) with the occurrence of ankylosing spondylitis (AS), and the effects of Bushen Tongdu Decoction (BTD) on serum TNF-alpha and TGF-beta1, levels in patients.</p><p><b>METHODS</b>Seventy five AS patients were assigned to two groups, 30 in the group I treated with Azulfidine and35 in the group II treated with BTD. ELISA was adopted to detect the levels of TNF-alpha and TGF-beta1, in patients before and after 24-week treatment. Meantime, erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) were measured as well. Besides, the levels of TNF-alpha and TGF-beta1 in 30 healthy persons were also detected for normal control.</p><p><b>RESULTS</b>Before treatment, as compared with normal control, the TNF-alpha increased and TGF-beta1 decreased (P < 0.01) in the two treatment groups; the serum level of TNF-alpha was positively correlated with ESR and CRP (r = 0.296, r = 0.249; P < 0.05), but the serum level of TGF-beta1 showed no correlation with them (r = -0.222, r = -0.203; P > 0.05). After treatment, TGF-beta1 increased, while TNF-alpha, CRP and ESR decreased in group II (P < 0.01).</p><p><b>CONCLUSION</b>BTD can regulate TNF-alpha and TGF-beta1 levels to suppress the immune inflammatory reaction so as to treat AS.</p>

Screening of new HIV inhibitors based on the database of traditional Chinese medicine / 药学学报

<p><b>AIM</b>To report the preliminary result of the HIV inhibitor screening based on cheminformatics tools and the traditional Chinese medicine database.</p><p><b>METHODS</b>Database search was carried out with saquinavir molecule as a template, further screening was made with docking. Detailed studies using molecular dynamics simulation of 50 ps and 200 ps were made with respect to a potential leading compound, leucovorin.</p><p><b>RESULTS</b>The leucovorin molecule distinguished from other molecules as a potential drug candidate and is subject to extensive studies. The bonding profile and energy were calculated with MD simulations.</p><p><b>CONCLUSION</b>Our results could be very helpful when we modify leucovorin or design new inhibitors against HIV.</p>

Expression and subcellular localization of P9-ZFD protein in patients with myasthenia gravis / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To express and purify the protein coded by the TRAF-type zinc finger domain of myasthenia gravis (MG)-related gene P9 (P9-ZFD) and to prepare P9-ZFD antiserum for detecting expression and subcellular distribution of P9-ZFD protein in the skeletal muscles of patient with MG.</p><p><b>METHODS</b>The cDNA encoding P9-ZFD was amplified by RT-PCR. The cloned P9-ZFD cDNA was ligated into pET24a, and the P9-ZFD recombinant protein was induced via E. coli. BL21 (DE3) and purified by histidine affinity chromatography. P9-ZFD antiserum was prepared and its titer and specificity were determined by ELISA and Western blot. Expression and subcellular distribution of P9-ZFD protein in the skeletal muscles of MG and control were studied.</p><p><b>RESULTS</b>The molecular weight of purified P9-ZFD protein was about 30 kD. Its purity was more than 95%. Antiserum specific for P9-ZFD was excellent. P9-ZFD protein is fully confined to the cytoplasm membrane of skeletal muscle cell of MG, obvious immunostaining was absent in the A, I, and Z bands of cytoplasm and no immunoreactivity was observed in the skeletal muscle cell of control.</p><p><b>CONCLUSION</b>P9-ZFD protein is expressed as a cytoplasm membrane-bound protein and has obvious distribution difference in the skeletal muscle cells of patient with MG and normal control.</p>