Capsaicin functions as a selective degrader of STAT3 to enhance host resistance to viral infection.

Although STAT3 has been reported as a negative regulator of type I interferon (IFN) signaling, the effects of pharmacologically inhibiting STAT3 on innate antiviral immunity are not well known. Capsaicin, approved for the treatment of postherpetic neuralgia and diabetic peripheral nerve pain, is an agonist of transient receptor potential vanilloid subtype 1 (TRPV1), with additional recognized potencies in anticancer, anti-inflammatory, and metabolic diseases. We investigated the effects of capsaicin on viral replication and innate antiviral immune response and discovered that capsaicin dose-dependently inhibited the replication of VSV, EMCV, and H1N1. In VSV-infected mice, pretreatment with capsaicin improved the survival rate and suppressed inflammatory responses accompanied by attenuated VSV replication in the liver, lung, and spleen. The inhibition of viral replication by capsaicin was independent of TRPV1 and occurred mainly at postviral entry steps. We further revealed that capsaicin directly bound to STAT3 protein and selectively promoted its lysosomal degradation. As a result, the negative regulation of STAT3 on the type I IFN response was attenuated, and host resistance to viral infection was enhanced. Our results suggest that capsaicin is a promising small-molecule drug candidate, and offer a feasible pharmacological strategy for strengthening host resistance to viral infection.

Clinical value of ultrasound-guided radiofrequency ablation in the treatment of retroperitoneal tumors / 中华肿瘤杂志

Objective: To explore the clinical value of ultrasound-guided radiofrequency ablation in the treatment of retroperitoneal tumors. Methods: The clinical data of 13 patients with retroperitoneal tumors treated with ultrasound-guided radiofrequency ablation in the First Affiliated Hospital of Zhengzhou University from January 2018 to January 2020 were analyzed retrospectively. The ablation effect was evaluated and the postoperative complications were observed. The changes of tumor volume before and after radiofrequency ablation were compared. Results: The symptoms of pain and dyspepsia were significantly improved after radiofrequency ablation, and the hospital stay was (9.2±2.9) days. The tumor was ablated completely in 10 cases, tumor residual in 1 case and tumor metastasis in 2 cases. One patient had postoperative duodenal perforation complicated with intra-abdominal infection, and no serious complications occurred in other patients. There were 20 lesions in 13 patients. The maximum diameter of 20 lesions before operation and 1, 3, 6 months after operation were (39.5±15.9) mm, (30.6±4.9)mm, (15.6±7.7) mm and (9.9±3.1) mm, respectively, the maximum diameters of 1, 3 and 6 months after operation were smaller than that before operation (P<0.05). Conclusion: Ultrasound-guided radiofrequency ablation is a real-time, accurate, safe and effective minimally invasive treatment with few complications, and has a high clinical value for retroperitoneal tumors.

Simplified plasmid cloning with a universal MCS design and bacterial in vivo assembly.

BACKGROUND: The ability to clone DNA sequences quickly and precisely into plasmids is essential for molecular biology studies. The recent development of seamless cloning technologies has made significant improvements in plasmid construction, but simple and reliable tools are always desirable for time- and labor-saving purposes. RESULTS: We developed and standardized a plasmid cloning protocol based on a universal MCS (Multiple Cloning Site) design and bacterial in vivo assembly. With this method, the vector is linearized first by PCR (Polymerase Chain Reaction) or restriction digestion. Then a small amount (10 ~ 20 ng) of this linear vector can be mixed with a PCR-amplified insert (5× molar ratio against vector) and transformed directly into competent E. coli cells to obtain the desired clones through in vivo assembly. Since we used a 36-bp universal MCS as the homologous linker, any PCR-amplified insert with ~ 15 bp compatible termini can be cloned into the vector with high fidelity and efficiency. Thus, the need for redesigning insert-amplifying primers according to various vector sequences and the following PCR procedures was eliminated. CONCLUSIONS: Our protocol significantly reduced hands-on time for preparing transformation reactions, had excellent reliability, and was confirmed to be a rapid and versatile plasmid cloning technique. The protocol contains mostly mixing steps, making it an extremely automation-friendly and promising tool in modern biology studies.

Gastrointestinal Symptoms Associated With Unfavorable Prognosis of COVID-19 Patients: A Retrospective Study.

Background and Aim: The global pandemic of COVID-19 has posed an enormous threat to the economy and people's lives across various countries. Patients with COVID-19 most commonly present with respiratory symptoms. However, gastrointestinal (GI) symptoms can also occur. We aimed to study the relationship between GI symptoms and disease prognosis in patients with COVID-19. Methods: In a single-center and retrospective cohort study, the outcomes in COVID-19 patients with or without GI symptoms were compared. The propensity score is a conditional probability of having a particular exposure (COVID-19 patients with GI symptoms vs. without GI symptoms) given a set of baseline measured covariates. Survival was estimated using the Kaplan-Meier method, and any differences in survival were evaluated with a stratified log-rank-test. To explore the GI symptoms associated with ARDS, non-invasive ventilator treatment, tracheal intubation, tracheotomy, and CRRT, univariable and multivariable COX regression models were used. Results: Among 1,113 eligible patients, 359 patients with GI symptoms and 718 without GI symptoms had similar propensity scores and were included in the analyses. Patients with GI symptoms, as compared with those without GI symptoms, were associated with a similar risk of death, but with higher risks of ARDS, non-invasive mechanical ventilation in COVID-19 patients, respectively. Conclusions: The presence of GI symptoms was associated with a high risk of ARDS, non-invasive mechanical ventilation and tracheal intubation in patients with COVID-19 but not mortality.

Red nucleus IL-6 mediates the maintenance of neuropathic pain by inducing the productions of TNF-α and IL-1ß through the JAK2/STAT3 and ERK signaling pathways.

We previously reported that interleukin (IL)-6 in the red nucleus (RN) is involved in the maintenance of neuropathic pain induced by spared nerve injury (SNI), and exerts a facilitatory effect via Janus-activated kinase 2/signal transducer and activator of transcription 3 (JAK2/STAT3) and extracellular signal-regulated kinase (ERK) signal transduction pathways. The present study aimed at investigating the roles of tumor necrosis factor-α (TNF-α) and IL-1ß in RN IL-6-mediated maintenance of neuropathic pain and related signal transduction pathways. Being similar to the elevation of RN IL-6 three weeks after SNI, increased protein levels of both TNF-α and IL-1ß were also observed in the contralateral RN three weeks after the nerve injury. The upregulations of TNF-α and IL-1ß were closely correlative with IL-6 and suppressed by intrarubral injection of a neutralizing antibody against IL-6. Administration of either the JAK2 antagonist AG490 or the ERK antagonist PD98059 to the RN of rats with SNI remarkably increased the paw withdrawal threshold (PWT) and inhibited the up-regulations of local TNF-α and IL-1ß. Further experiments indicated that intrarubral injection of exogenous IL-6 in naive rats apparently lowered the PWT of the contralateral hindpaw and boosted the local expressions of TNF-α and IL-1ß. Pretreatment with AG490 could block IL-6-induced tactile hypersensitivity and suppress the up-regulations of both TNF-α and IL-1ß. However, injection of PD98059 in advance only inhibited the upregulation of IL-1ß, but not TNF-α. These findings indicate that RN IL-6 mediates the maintenance of neuropathic pain by inducing the productions of TNF-α and IL-1ß. IL-6 induces the expression of TNF-α through the JAK2/STAT3 pathway, and the production of IL-1ß through the JAK2/STAT3 and ERK pathways.

Prolyl 4-hydroxylase subunit alpha 3 presents a cancer promotive function in head and neck squamous cell carcinoma via regulating epithelial-mesenchymal transition.

OBJECTIVE: This study aimed to explore the prognostic value and functional role of Prolyl 4-Hydroxylase Subunit Alpha 3 (P4HA3) in head and neck squamous cell carcinoma. METHODS: We downloaded the RNA-Seq dataset of head and neck squamous cell carcinoma for analyzing the expression of P4HA3 and determining its prognostic value. Head and neck squamous cell carcinoma cell lines CAL27 and FaDu were chose for gain- and loss-of-function of P4HA3 tests. mRNA and protein levels of P4HA3 in head and neck squamous cell carcinoma cells were tested by quantitative real-time PCR and western blot respectively. Cell counting Kit-8, clone formation assay, Transwell assay were used to determine the effect of P4HA3 on the proliferative, invasive and migratory capacities of head and neck squamous cell carcinoma cells. RESULTS: Bioinformatics analysis showed that P4HA3 was up-regulated in head and neck squamous cell carcinoma tissues, and had an independent prognostic value for head and neck squamous cell carcinoma patients. The outcomes of gain- and loss-of-function tests illustrated that P4HA3 significantly boosted head and neck squamous cell carcinoma cell proliferative, invasive and migratory abilities. Besides, western blot assay demonstrated that P4HA3 could remarkably activate the epithelial-mesenchymal transition process, with reduced the level of E-cadherin and up-regulated the levels of N-cadherin, Vimentin and Snail. CONCLUSION: We deduce that P4HA3 acts as an oncogene that raises tumor cell viability and metastasis partly by inducing the epithelial-mesenchymal transition process, suggesting that P4HA3 may be considered as a valuable biomarker for head and neck squamous cell carcinoma treatment.

Prevalence of urogenital tract infections with Ureaplasma urealyticum or HPV among men of reproductive age and the associated factors / 中华男科学杂志

Objective@#To investigate the prevalence of urogenital tract infections with Ureaplasma urealyticum (UU) and human papilloma virus (HPV) in males of reproductive age and the associated factors.@*METHODS@#Using the multi-stage cluster sampling method and a structured questionnaire, we conducted an investigation among 18－50 years old males in Songjiang District, Shanghai, from August 2016 to July 2018. We collected secretory specimens from the urogenital tract of the subjects and detected the infections of UU and HPV by laboratory examination.@*RESULTS@#Among the 621 males included in this study, 279 (44.93%) were found infected with UU, 18 (2.90%) with HPV, and 15 (2.42%) with both UU and HPV. Univariate analysis showed that smokers had a higher rate of UU infection (50.54% ［140/277］) than non-smokers (40.41 ［139/344］), and those with senior high school or secondary technical school education had a higher rate of HPV infection (4.84% ［12/248］) than others (1.61% ［6/373］). Binary stepwise logistic regression analysis revealed a higher risk of UU infection in the subjects with junior high school or lower education than in others (OR = 0.61, 95% CI: 0.39－0.96) as well as in smokers than in non-smokers (OR = 1.46, 95% CI: 1.01－2.01).@*CONCLUSIONS@#The prevalence of UU infection is high, while that of HPV is low among men of reproductive age in Songjiang, Shanghai. The screening of UU infection should be enhanced among men of reproductive age, especially among smokers and those with lower education.

Therapeutic effects of transvaginal natural orifice transluminal endoscopic surgery sacrocolpopexy on pelvic organ prolapse / 中国实用妇科与产科杂志

OBJECTIVE: To study the efficacy of transvaginal natural orifice transluminal endoscopic surgery(vNOTES)sacrocolpopexy in the treatment of pelvic organ prolapse(POP).METHODS: From June 2017 to September 2018,33 POP patients who underwent vNOTES sacrocolpopexy in the Third Affiliated Hospital of Guangzhou Medical University were collected.The operative time,blood loss and postoperative complications were recorded to evaluate the safety of the operation.After regular follow-up,POP-Q was used to objectively evaluate the effect of surgery,and PFIQ-7 was used to evaluate the recovery effect.RESULTS: In this group,3 of 33 cases were converted to transumbilical laparoscopy,and in the remaining 30 cases the surgery was successfully completed.The operation time was(184.97±41.91)min,and mean estimated blood loss was(28.33±18.77)mL.No complications such as exposure or infection of mesh were found after operation.The POP-Q indicators were significantly improved in 1,3,6 months after surgery(P<0.05),and the quality of life was better after operation than before(P<0.05).CONCLUSION: vNOTES is a feasible and safe approach for sacrocolpopexy with positive short-term efficacy in the treatment of POP.

Bioinformatics Based Therapeutic Effects of Sinomenium Acutum / 中国结合医学杂志

OBJECTIVE@#To decipher the possible mechanisms of Sinomenium Acutum (SA) in treating diseases by a bioinformatics method.@*METHODS@#SA ingredients were searched according to Chinese Pharmacopoeia, Chinese Medicine Dictionary and Traditional Chinese Medicines Database (TCMD). Active compounds and target proteins of SA were acquired through the Pubchem platform. Pathway, network and function analyses of SA were performed with ingenuity pathway analysis (IPA), a bioinformatics analysis platform. Disease, biofunction-target networks were established with Cytoscape.@*RESULTS@#Eighteen ingredients from SA were obtained. Seven active ingredients with 31 active target proteins were acquired according to PubChem Bioassay test. By IPA analysis, 277 canonical pathways belonging to 17 function categories were collected, 23 kinds of diseases, 21 categories bio-functions were obtained. Based on P value, calculated by IPA, the top 5 significant pathway of SA targets include phosphatidylinositol 3 kinase/Akt (PI3K/Akt) signaling, prostate cancer signaling, macrophage migration inhibitory factor (MIF) regulation of innate immunity, Guanosine-binding protein coupled receptor (GPCR) signaling, and ataxia telangiectasia mutated protein (ATM) signaling. Disease and bio-function network analysis indicated that mitogen activated protein kinase 1 (MAPK1), MAPK3, p65 nuclear factor κB (RELA), nuclear factor of κB inhibitor alpha (NFκBIA), interleukin 1β(IL-1β), prostaglandin G/H synthase 2 (PTGS2) and tumor protein 53 (TP53) were the critical targets in various diseases treated by SA.@*CONCLUSION@#In the different view of target, pathway, disease and bio-function, inflammation was found to be a central theme in many chronic conditions. SA could be used not only as an anti-inflammatory agent, but also for the treatment of cancers, neurological diseases, psychological disorders and metabolic diseases.

[A reduction technique for the treatment of refractory femoral trochanteric fracture].

OBJECTIVE: To explore the effect of periosteal stripper through spiral blade incision combined with lateral sight bracket prying method in the treatment of irreducible femoral trochanteric fractures with proximal upwarping of fracture. METHODS: From March 2015 to October 2017, 26 patients with refractory femoral trochanteric fractures with proximal upwarp of fracture were treated by periosteal dissection through a spiral blade incision and lateral sight bracket prying, included 9 males and 17 females with an average age of (76.3±8.4) years old ranged from 63 to 85 years old. According to OA/OTA classification, 5 cases were type A1.3 fracture, 6 cases were A2.1, 7 cases were A2.2, 8 cases were A2.3. All cases were closed fractures. The amount of bleeding during operation and the time of fracture healing were observed and recorded. The Harris score of the hip joint function was performed. RESULTS: All cases were followed up from 7 to 26 months with an average of (17.4±4.7) months. Bone union time was from 9 to 15 months with an average of(12.4±3.5)weeks according to X-rays. At the final follow-up, Harris scores were 86 to 93 points with an average of (85.8±5.6) points, 23 cases were classified as excellent, 2 as good, and 1 as fair. CONCLUSIONS: Using periosteum stripper through spiral blade incision combined lateral sight bracket prying for the treatment of irreducible femoral trochanteric fracture, the reduction effect is accurate, the operation time is shortened obviously, no auxiliary small incision is needed, the clinical effect is good, the reduction technique is simple and practical.

The status and tendency projection of contraceptive status in China,Japan, India, South Africa and the United States of America from 1970 to 2030 / 复旦学报（医学版）

Objective To provide a basis to satisfy the need for family planning of women of reproductive age,according to the analysis and estimation of family planning status of married or in union women of reproductive age among China,Japan,India,South Africa and the United States of America.Methods U test and Chi-square test were applied to analyze the current situation (1970-2016) and tendency (2017-2030) of family planning indicators of the five countries from 1970 to 2030,which were selected from open database of World Contraceptive Use 2016.Results Compared with the family planning status in 1970,there was a significant change in total demand for family planning,contraceptive prevalence and unmet need for family planning (P＜0.001).To be specific,in 2016,contraceptive prevalence of married or in-union women of reproductive age in China,Japan,India,South Africa and the United States of America was 83.35％,59.9％,56.4％,64.9％ and 73.7％,respectively;unmet need for family planning was 3.8％,13.1％,15.5％,12.2％ and 7.2％,respectively.Besides,contraceptive methods composition in the five countries were changed in 2016 (P＜0.001) and the prevalence of modern methods was higher than traditional methods.Meanwhile,the percentage of demand for family planning satisfied with modern methods were increasing from 1970 to 2016.On the other hand,the projections of total demand for family planning,contraceptive prevalence,unmet need for family planning,composition of contraceptive methods and unmet need for modern methods would change from 2017 to 2030 (P＜0.001).It was estimated that,in 2030,China would have the largest number of people using contraceptive methods and India would have the largest number of people with unmet need for family planning.Moreover,China and India might be the countries with most users of modern methods,and India might be associated with the most users of traditional methods.Conclusions Considering current and projected status of huge total demand for family planning,high contraceptive prevalence,low unmet need for family planning,it is essential to supply approximate contraceptive methods.

Human papillomavirus infection and male reproductive health / 中华男科学杂志

Human papillomavirus (HPV) infection is one of the worldwide sexually transmitted diseases (STD), and the DNA of HPV can exist in the normal epithelium of reproductive organs of both men and women. Because the majority of HPV infections are asymptomatic, healthy HPV-carriers become the main source of the infection. Studies show that HPV infection in men is correlated with STD, infertility, tumor of reproductive organs, and infection in the sexual partners. Therefore, measures should be taken to reduce male HPV infection, including circumcision, fewer sexual partners, and condom use.

Antibody-drug conjugates:design and clinical progress / 药学学报

Antibody-drug conjugates, constructed with monoclonal antibodies, linker and cytotoxins, have distinctive advantages over chemotherapy drugs and antibody drugs in cancer therapy. In this review, the strategy of developing ADCs, and the important progress in past decade are well summarized. The representative ADCs in the pipeline are introduced and characterized with their new features. While, perspective for future directions of ADCs is proposed.

Progress in the study of near-infrared fluorescent probes for the detection of β-amyloid deposition in Alzheimer's disease / 药学学报

Alzheimer's disease (AD) is the most common cause of cognitive impairment in older people. With the aging of society is more and more serious, AD caused great burden to patients and society. A β is a classical biomarker of AD, which has been widely used in clinical diagnosis of AD patients. Compared with positron emission tomography (PET) and single photon emission computed tomography (SPECT), near infrared fluorescence imaging has many advantages including highly sensitive, non-invasive, safety and inexpensive. Therefore, many research groups have focused on developing the molecular probes of near-infrared fluorescence (NIRF) imaging. In this article, we will review the progress of the probes of NIRF.

Outcome and prognosis in acute myeloid leukemia patients treated with related peripheral blood hematopoietic stem cell transplantation / 中国实验血液学杂志

This study was purposed to evaluate the outcome of acute myeloid leukemia patients treated with related peripheral blood hematopoietic stem cell transplantation (PBHSCT) and analyse the potential prognostic factors. A total of 64 acute myeloid leukemia patients treated with related peripheral allo-HSCT from march 2008 to august 2012 in our hospital were enrolled in the analysis. All the patients received either HLA-matched related or mismatched related donor mobilized peripheral blood stem cells. All the patients were followed up and evaluated for overall survival (OS), leukemia-free survival (LFS) and relapse rate (RR) , and the potential prognostic factors well analyzed. The results showed that the 3 year OS , LFS and RR were 61.9%, 52% and 39.1% respectively. Univarite analysis demonstrated that the disease status before transplantation (P < 0.01) , donor type (P < 0.01), white blood cell count at initial diagnosis (P < 0.05) are related with outcome, and severe aGVHD has some influence on the outcome (P > 0.05) . Multivariate analysis indicated the status of disease before transplantation, donor type, severe aGVHD are the most important prognostic factors. It is concluded that the related PBHSCT is effective treatment method for AML patients, recurrence is the main reason for the failure after transplantation, disease status before transplantation, donor type, and severe aGVHD are independent prognostic factors.

AZD1480 can inhibit the biological behavior of ovarian cancer SKOV3 cells in vitro.

OBJECTIVE: To study the mechanism of effects of AZD1480 on the SKOV3 ovarian cancer cell line. METHODS: The MTT method was used to assess cellular proliferation, flow cytometry for cellular apoptosis, the scratch test to determine migration, transwell chamber assays to detect cellular invasion, plate clone experiments to detect the clone forming ability and Western blotting to determine p-STAT3 protein levels. RESULTS: The proliferation rate, migration ability, invasiveness and the clone forming ability of SKOV3 cells were reduced after treatment with AZD1480, while apoptosis rate and chemotherapeutic susceptibility were increased. After treatment with AZD1480 plus cisplatin, the apoptosis rate increased significantly while the expression level of p-STAT3 protein was decreased. CONCLUSION: AZD1480 can inhibit the proliferation, invasion, metastasis and clone formation of SKOV3 cells, induce cellulsar apoptosis, increase the chemotherapeutic sensitivity and reduce the expression level of p-STAT3 protein.

The influence of high birth weight on the blood pressure during childhood—a cohort study / 中华流行病学杂志

Objective To explore the effects of high birth weight (HBW) on blood pressure (BP) during childhood.Methods A total of 1435 couples with high or normal birth weight were selected from a birth cohort who were born between 1993 and 1995 in Wuxi,China and,followed between 2005 and 2007.A questionnaire survey was conducted,with their height,weight and blood pressure measured.Results The medians of systolic blood pressure (SBP) and diastolic BP (DBP)of HBW group were 100 mm Hg (Q1-Q3∶90-110 mm Hg),64 mm Hg (Q1-Q3∶60-70 mm Hg) in the high birth weight group,respectively,while they were 100 mm Hg (Q1-Q3∶90-108 mm Hg),62 mm Hg (Q1-Q3∶60-70 mm Hg) in the normal weight group.No statistical significant difference was found between the exposed and unexposed groups.The proportion of elevated BP,including prehypertension and hypertension,in exposed group was 13.66％,a bit higher than that (11.57％) of the unexposed group (P ＝ 0.055).The relative risk (RR) of HBW on elevated BP was 1.06 (0.92-1.21),after adjusted the following parameters as BMI in childhood,sex,age during the follow-up period,age of mother at delivery,gestational age,type of feeding at infancy,dietary habit as well as the frequencies of exercise during childhood.Conclusion No statistically significant correlation was found between high birth weight and blood pressure of children.

The influence of smoking on the routine parameters of semen quality / 中华流行病学杂志

Objective To explore the effect and heaviness of smoking on the routine parameters of semen quality.Methods A total of 727 eligible subjects including 421 smokers and 306 nonsmokers were selected in 2004-2006 in Hebei,Shanxi,Guizhou,Zhejiang and Shandong provinces.Data on background information,general demographic characteristics,exposure to smoking and related confounding factors were obtained through a questionnaire survey.Semen samples of all the subjects were collected during the period of study.Parameters of semen were examined according to the manual recommended by WHO.Results Four models from the multiple logistic regression regarding the semen routine parameters were used as the dependent variables to be fitted in descending order respectively,while six confounders including area,age,abstinence time,education level,history of alcohol intake and the exposure to pesticides atc.,were adjusted.Results showed that smoking could affect the sperm viability,with significantly higher proportion of abnormal sperm viability seen in the smoking group (OR= 1.77,95％ CI:1.12-2.80).Proportions regarding the abnormal mobility and morphology of sperms in the smoking group were also significantly higher than in the control group,with OR values as 1.51 (95％CI:1.00-2.27) and 2.55 (95％CI:1.68-3.88) respectively.The density of sperms was significantly decreased in the smoking groups who smoked more than 20 cigarettes per day (OR= 1.70,95％CI:1.05-2.76) or the total numbers of cigarette consumption exceeding 150 000(OR=1.84,95％CI:1.03-3.29).Conclusion Smoking showed adverse effects on the quality of semens,especially for heavy or long-term smokers.

Detection of bone marrow involvement by lymphoma cells in patients with B-NHL by using flow cytometry analysis / 中国实验血液学杂志

This study was purposed to investigate the significance of using (FCM) flow cytometry for detection bone marrow involvement by lymphoma cells in untreated patients with B cell non-Hodgkin's lymphoma (B-NHL). Bone marrow samples of 54 patients with B-NHL were analyzed by flow cytometry, morphological method and molecular biology technique. Bone marrow involvement was diagnosed based on the results of morphology, FCM and molecular biology. The results indicated that the positive ratios of bone marrow involvement were different detected by three methods, the most sensitive method was FCM (positive rate was 27.5%), the moderately sensitive method was molecular biology (positive rate was 22.2%), the inferior method was morphology method (positive rate was 5.6%). Combined with three methods, the positive rate increased to 38.9%. The monoclonal B lymphocytes were detected by FCM in fourteen patients, the ratios of kappa light chain/lambda light chain were far beyond the diagnostic threshold. Even in patients with early stage, lymphoma cells still could be detected by FCM in involved bone marrow. Monoclonal B lymphocytes were detected by FCM in three patients who were diagnosed bone marrow involvement by morphological method. The concordance between FCM and molecular biology was relatively high (70.4%, p=0.14). It is concluded that the detection of monoclonal B lymphocytes by using FCM has high sensitivity and accuracy in patients with B-NHL. Evaluation whether the bone marrow has been involved by lymphoma cells should be recommend to every patient with B-NHL before chemotherapy and every disease stages. Combined application of FCM and molecular biology method would enhance diagnostic efficacy.

Tissue-specific expression of the PNZIP promoter is mediated by combinatorial interaction of different cis-elements and a novel transcriptional factor.

Recent studies demonstrated that PNZIP and its homologs encode a special cyclase and play an important role in chlorophyll biosynthesis in higher plants. To investigate the molecular mechanism governing the PNZIP gene, the PNZIP promoter was isolated and analyzed. Deletion analysis indicated that G-box is an important element in the regulation of the reporter gene expression. Further mutation assay demonstrated that G-box and GATACT elements are necessary and sufficient for the high and tissue-specific expression of the GUS gene. Using yeast one-hybrid screening, we have isolated a novel tobacco bZIP protein, NtbZIP, which can specifically recognize the G-box of the PNZIP promoter. The NtbZIP protein shares a limited amino acid homology to Arabidopsis ABI5 and AtAREB1 and very low homology to other bZIP proteins. Northern blot analysis showed that the NtbZIP gene is not induced by exogenous ABA and is expressed in different tobacco organs. Cotransformation assays showed that the NtbZIP protein could activate the transcription of the GUS gene driven by the PNZIP promoter. Transgenic tobaccos analysis demonstrated that constitutively expressing antisense NtbZIP gene resulted in a lower NTZIP synthesis and reduced chlorophyll levels. We suggest that NTZIP is a target gene of NtbZIP, which is involved in the regulation of chlorophyll biosynthesis.