Research progress and prospects for the use of plant metabolomics in quality evaluation of traditional Chinese medicinal materials / 药学学报

The quality of traditional Chinese medicine has a direct impact on the effectiveness and safety of its use, and is the premise necessary to ensure the healthy development of the traditional Chinese medicine industry. Comprehensive and accurate control and evaluation of the quality of medicinal materials is of great significance to the traditional Chinese medicine industry, but the complexity and dynamics of the chemical composition of medicinal materials makes their quality evaluation a challenge. Plant metabolomics provides an integrated and comprehensive analysis that is consistent with the holistic approach of traditional Chinese medicine. Chemical information therein promotes the establishment of a traceable system and provides new ideas and methods for the quality evaluation of medicinal materials. Plant metabolomics in the quality evaluation of medicinal materials is gradually increasing, and the core is the screening and identification of differential metabolites or specific marker compounds by means of stoichiometry. This study focused on the main factors that affect the quality of medicinal materials, such as origin, environmental adversity, varieties, harvest time, commercial specification and TCM processing. We describe the research progress in plant metabolomics combined with chemometrics analysis for the quality control and evaluation of medicinal materials, summarize existing problems, identify trends, and propose future research directions. Metabolomics plays an increasingly important role in the quality evaluation of medicinal materials, but the absolute qualitative and quantitative information of metabolomics needs to be further developed, and a single 'omics' technique is not sufficient for an in-depth analysis of medicinal value. In the future, standardization of plant metabolomics methods and a more complete database should be actively promoted, and plant metabolomics should be integrated into quality marker exploration. Plant metabolomics will need to be integrated with other 'omics' methods to improve the quality and evaluation system of medicinal materials.

Involvement of Opioid Peptides in the Analgesic Effect of Spinal Cord Stimulation in a Rat Model of Neuropathic Pain / 神经科学通报·英文版

Spinal cord stimulation (SCS)-induced analgesia was characterized, and its underlying mechanisms were examined in a spared nerve injury model of neuropathic pain in rats. The analgesic effect of SCS with moderate mechanical hypersensitivity was increased with increasing stimulation intensity between the 20% and 80% motor thresholds. Various frequencies (2, 15, 50, 100, 10000 Hz, and 2/100 Hz dense-dispersed) of SCS were similarly effective. SCS-induced analgesia was maintained without tolerance within 24 h of continuous stimulation. SCS at 2 Hz significantly increased methionine enkephalin content in the cerebrospinal fluid. The analgesic effect of 2 Hz was abolished by μ or κ opioid receptor antagonist. The effect of 100 Hz was prevented by a κ antagonist, and that of 10 kHz was blocked by any of the μ, δ, or κ receptor antagonists, suggesting that the analgesic effect of SCS at different frequencies is mediated by different endorphins and opioid receptors.

Systematic evaluation and sequential Meta-analysis of modified Xuefu Zhuyu Decoction combined with Western medicine in treatment of leiomyoma / 中国中药杂志

CNKI, PubMed and other databases were retrieved to extract eligible randomized controlled trial(RCT) about modified Xuefu Zhuyu Decoction(MXZD) combined with Western medicine(trial group) versus Western medicine alone(control group) in the treatment of leiomyoma. Therefore, a total of 25 RCTs were included, involving 2 328 patients. Bias risk evaluation tool in Cochrane Handbook 5.1.0 was used for evaluating the quality of these RCTs. Meta-analysis was performed for the reported indicators, including total efficiency, serum hormone level [progesterone(P), luteinizing hormone(LH), estradiol(E_2), follicle stimulating hormone(FSH)], uterine size, fibroids size and adverse reactions by using Stata 14.0 software. Meta-analysis showed that the total efficiency(RR=1.21,95%CI[1.17,1.25],P<0.05) of trial group was better than that of control group. Serum hormone level(WMD_P=-3.86,95%CI[-4.31,-3.41],P<0.05; WMD_(LH)=-3.64,95%CI[-4.47,-2.82],P<0.05; WMD_(E_2)=-39.99,95%CI[-53.45,-26.52],P<0.05; WMD_(FSH)=-3.79,95%CI[-4.86,-2.72],P<0.05), uterine size(WMD=-50.02,95%CI[-55.98,-44.06],P<0.05), fibroids size(WMD=-15.79,95%CI[-18.11,-13.46],P<0.05) and adverse reactions(RR=0.65,95%CI[0.48,0.88],P<0.05) of trial group were all lower than those of control group, with statistical significances. Trial sequential analysis(TSA) was performed by using TSA 0.9 software, and showed a reliable therapeutic effect of the experimental group. In short, our study indicated that modified Xuefu Zhuyu Decoction combined with Western medicine had a better therapeutic effect on leiomyoma than Western medicine alone, but more high-quality studies are needed to verify this conclusion in the future.

Responses of Primary Afferent Fibers to Acupuncture-Like Peripheral Stimulation at Different Frequencies: Characterization by Single-Unit Recording in Rats / 神经科学通报·英文版

The pain-relieving effect of acupuncture is known to involve primary afferent nerves (PANs) via their roles in signal transmission to the CNS. Using single-unit recording in rats, we characterized the generation and transmission of electrical signals in Aβ and Aδ fibers induced by acupuncture-like stimuli. Acupuncture-like signals were elicited in PANs using three techniques: manual acupuncture (MAc), emulated acupuncture (EAc), and electro-acupuncture (EA)-like peripheral electrical stimulation (PES). The discharges evoked by MAc and EAc were mostly in a burst pattern with average intra-burst and inter-burst firing rates of 90 Hz and 2 Hz, respectively. The frequency of discharges in PANs was correlated with the frequency of PES. The highest discharge frequency was 246 Hz in Aβ fibers and 180 Hz in Aδ fibers. Therefore, EA in a dense-disperse mode (at alternating frequency between 2 Hz and 15 Hz or between 2 Hz and 100 Hz) best mimics MAc. Frequencies of EA output >250 Hz appear to be obsolete for pain relief.

Application of n-dodecane as an oxygen vector to enhance the activity of fumarase in recombinant Escherichia coli: role of intracellular microenvironment

Abstract The effect of the intracellular microenvironment in the presence of an oxygen vector during expression of a fusion protein in Escherichia coli was studied. Three organic solutions at different concentration were chosen as oxygen vectors for fumarase expression. The addition of n-dodecane did not induce a significant change in the expression of fumarase, while the activity of fumarase increased significantly to 124% at 2.5% n-dodecane added after 9 h induction. The concentration of ATP increased sharply during the first 6 h of induction, to a value 7600% higher than that in the absence of an oxygen-vector. NAD/NADH and NADP/NADPH ratios were positively correlated with fumarase activity. n-Dodecane can be used to increase the concentration of ATP and change the energy metabolic pathway, providing sufficient energy for fumarase folding.

Clinical analysis of 50 cases of serous cavity effusion related with paragonimiasis / 中国血吸虫病防治杂志

To analyze the clinical characteristics of serous cavity effusion related with paragonimiasis, so as to improve the physician’s ability of the clinical diagnosis and treatment.The clinical data of 50 cases of serous cavity effusion related with paragonimiasis diagnosed in a hospital in recent 3 years were collected and retrospectively analyzed.Among the 50 patients, there were 35 males and 15 females, and 35 children and 15 adults. Eighteen patients had a clear history of exposure to the foci. Among the children, the respiratory symptoms accounted for 68.6% (24/35), gastrointestinal symptoms for 22.9% (8/35), and no apparent symptoms for 8.6% (3/35); among the adults, the respiratory symptoms accounted for 93.3% (14/15) and the abdominal distention for 6.7% (1/15). The paragonimus antibody was positive in all the patients. The chest radiography or CT showed pulmonary inflammatory exudation and serous cavity effusion. Eosinophilia in blood was found in 25 cases, a large number of eosinophils in serous cavity effusion were found under microscopy in 12 cases, and eosinophil infiltration of pleura was found in 7 cases. All were cured after the patients received chemotherapy (praziquantel) and drainage fluid.The clinical manifestations of paragonimiasis are not very specific and paragonimiasis is often accompanied with serous cavity effusions. Clinically, paragonimiasis is easily misdiagnosed as tuberculosis or cancer. We should consider about the possibility of paragonimiasis when eosinophil is high in blood, serous cavity effusion and pleura.

Development of an Autism Subtyping Questionnaire Based on Social Behaviors / 神经科学通报·英文版

Autism spectrum disorder can be differentiated into three subtypes (aloof, passive, and active-but-odd) based on social behaviors according to the Wing Subgroups Questionnaire (WSQ). However, the correlations between the scores on some individual items and the total score are poor. In the present study, we translated the WSQ into Chinese, modified it, validated it in autistic and typically-developing Chinese children, and renamed it the Beijing Autism Subtyping Questionnaire (BASQ). Our results demonstrated that the BASQ had improved validity and reliability, and differentiated autistic children into these three subtypes more precisely. We noted that the autistic symptoms tended to be severe in the aloof, moderate in the passive, and mild in the active-but-odd subtypes. The modified questionnaire may facilitate etiological studies and the selection of therapeutic regimes.

Lumbopelvic Fixation and Sacral Decompression for U-shaped Sacral Fractures: Surgical Management and Early Outcome / 华中科技大学学报（医学）（英德文版）

U-shaped sacral fractures are rare and often difficult to diagnose primarily due to the difficulty in obtaining adequate imaging and the severe associated injuries.These fractures are highly unstable and frequently cause neurological deficits.The majority of surgeons have limited experience in management of U-shaped sacral fractures.No standard treatment protocol for U-shaped sacral fractures has been available till now.This study aimed to examine the management of U-shaped sacral fractures and the early outcomes.Clinical data of 15 consecutive patients with U-shaped sacral fracture who were admitted to our trauma center between 2009 and 2014 were retrospectively analyzed.Demographics,fracture classification,mechanism of injury and operative treatment and deformity angle were assessed.All the patients were treated with lumbopelvic fixation or (and) sacral decompression.EQ-5d score was applied to evaluate the patients' quality of life.Of the 15 consecutive patients with U-shaped sacral fracture,the mean age was 28.8 years (range:15-55 years) at the time of injury.There were 6 females and 9 males.The mean followup time was 22.7 months (range:9-47 months) and mean full weight-bearing time was 9.9 weeks (range:8-14 weeks).Ten patients received lumbopelvic fixation and sacral decompression,one lombosacral fixation,and 4 merely sacral decompression due to delayed diagnosis or surgery.The post-operation deformity angle (mean 27.87°,and range:8°-90°) of the sacrum was smaller than that pre-operation (mean 35.67;range:15-90) with no significance difference noted.At the latest follow-up,all patients obtained neurological recovery with different extents.Visual analogue score (VAS) was reduced from preoperative 7.07 (range:5-9) to postoperetive 1.93 (range:1-3).All patients could walk without any aid after treatment.Eight patients were able to care for themselves and undertook some daily activities.Five patients had returned to work full time.In conclusion,lumbopelvic fixation is an effective method for stabilization of U-shaped sacral fractures with fewer complications developed.Effective reduction and firm fixation are the prerequisite of early mobilization and neurological recovery.Sacral decompression effectively promotes neurological recovery even in patients with old U-shaped sacral fractures.

Lumbopelvic Fixation and Sacral Decompression for U-shaped Sacral Fractures: Surgical Management and Early Outcome / 华中科技大学学报（医学）（英德文版）

U-shaped sacral fractures are rare and often difficult to diagnose primarily due to the difficulty in obtaining adequate imaging and the severe associated injuries.These fractures are highly unstable and frequently cause neurological deficits.The majority of surgeons have limited experience in management of U-shaped sacral fractures.No standard treatment protocol for U-shaped sacral fractures has been available till now.This study aimed to examine the management of U-shaped sacral fractures and the early outcomes.Clinical data of 15 consecutive patients with U-shaped sacral fracture who were admitted to our trauma center between 2009 and 2014 were retrospectively analyzed.Demographics,fracture classification,mechanism of injury and operative treatment and deformity angle were assessed.All the patients were treated with lumbopelvic fixation or (and) sacral decompression.EQ-5d score was applied to evaluate the patients' quality of life.Of the 15 consecutive patients with U-shaped sacral fracture,the mean age was 28.8 years (range:15-55 years) at the time of injury.There were 6 females and 9 males.The mean followup time was 22.7 months (range:9-47 months) and mean full weight-bearing time was 9.9 weeks (range:8-14 weeks).Ten patients received lumbopelvic fixation and sacral decompression,one lombosacral fixation,and 4 merely sacral decompression due to delayed diagnosis or surgery.The post-operation deformity angle (mean 27.87°,and range:8°-90°) of the sacrum was smaller than that pre-operation (mean 35.67;range:15-90) with no significance difference noted.At the latest follow-up,all patients obtained neurological recovery with different extents.Visual analogue score (VAS) was reduced from preoperative 7.07 (range:5-9) to postoperetive 1.93 (range:1-3).All patients could walk without any aid after treatment.Eight patients were able to care for themselves and undertook some daily activities.Five patients had returned to work full time.In conclusion,lumbopelvic fixation is an effective method for stabilization of U-shaped sacral fractures with fewer complications developed.Effective reduction and firm fixation are the prerequisite of early mobilization and neurological recovery.Sacral decompression effectively promotes neurological recovery even in patients with old U-shaped sacral fractures.

Advantages and evidences research on Chinese medicine for treatment of pelvic inflammatory disease / 中国中药杂志

Pelvic inflammatory disease is an infectious disease. At present, Western medicine is mainly treated with antibiotics. However, the situation of antibiotics abuse is so grim that the potential risks such as the imbalance of bacteria, the resistance of bacteria, the production of super bacteria and the increase of adverse reactions are becoming more and more serious. Therefore, it is urgent to find a way to supplement or substitute antibiotics for the treatment of this disease. Traditional Chinese medicine treatment of the disease is effective and has its unique advantages. This paper mainly discusses the advantages and evidences of traditional Chinese medicine (TCM) treatment of pelvic inflammatory disease, to further prove the effectiveness and safety of TCM treatment and to provide medical evidence of reducing antibiotics use.

Treatment effect of Qi ming granule in preventing macular edema after phacoemulsification in diabetes / 国际眼科杂志(Guoji Yanke Zazhi)

AIM:To evaluate the clinical therapeutic efficacy of the Qi ming granule for macular edema ( ME ) in diabetic patients after phacoemulsification. METHODS:In this was a prospective clinical comparison study, 57 diabetic patients ( 76 eyes ) who underwent phacoemulsification were recruited and divided into two groups:treatment group (34 eyes) and control group (42 eyes) . All the patients in treatment group were given oral administration Qi ming granule (4. 5g, tid) and vitamin C ( 0. 1g, tid ) for 6mo postoperatively, while vitamin C ( 0.1g, tid ) for the controls. General clinical examinations, including blood glucose and glycated hemoglobin, as well as comprehensive standardized ophthalmic examinations were performed. Optical coherence tomography ( OCT ) were used to detect macular edema incidence and measure central field retinal thickness. RESULTS: No significant difference was found in the levels of blood glucose, glycated hemoglobin, course of disease, and macular thickness between the two groups during the initial visits. At the 6th month, 2 eyes ( 6%) eyes had clinically apparent macular edema in treatment group, while 6 ( 14%) eyes had clinically apparent macular edema in control group (P=0. 285). The central subfield retinal thickness values were significantly lower in the treatment group ( 211. 76±41. 21μm ) than those in control group (278. 36±48. 94μm) (P CONCLUSION:Qi ming granule can significantly reduce the incidences of macular edema and suppresses increasing retinal thickening after phacoemulsification in patients with diabetes mellitus.

Observation of serum protein fingerprinting in primary liver cancer patients of different Chinese medical syndromes before and after interventional treatment / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore changes of serum protein fingerprinting in primary liver cancer (PLC) patients of different Chinese medical syndromes before and after interventional treatment detected by surface enhanced laser desorption ionization time of flight mass spectrometry (SELDI-TOF-MS).</p><p><b>METHODS</b>Totally 154 PLC patients were assigned to 5 groups, i.e., Gan depression syndrome (GDS, 37 cases), Pi deficiency syndrome (PDS, 45 cases), dampness heat syndrome (DHS, 18 cases), blood stasis syndrome (BSS, 28 cases), yin deficiency syndrome (YDS, 26 cases). The mass spectra of serum proteins was analyzed by using SELDI-TOF-MS. Then the correlation between Chinese medical syndrome types and the mass spectra of serum proteins was explored before and after interventional treatment.</p><p><b>RESULTS</b>Compared with the healthy control group, the expression of serum proteins peak was down-regulated in GDS with M/Z being 6 589 and 4 182 Da, in DHS with M/Z being 5 710 Da, in YDS with M/ Z being 6 992 Da, while it was up-regulated in PDS with M/Z being 5 816 Da and in BSS with M/Z being 4 297 Da, showing statistical difference (P < 0.01). Compared with before intervention, the expression of serum proteins peak was down-regulated in GDS with M/Z being 6 589 and 4 182 Da, in PDS with M/Z being 5 816 Da, in DHS with M/Z being 5 710 Da in BSS with M/Z being 4 297 Da, while it was up-regulated in YDS with M/Z being 6 992 Da, showing statistical difference (P < 0.05, P < 0.01).</p><p><b>CONCLUSION</b>There was statistical difference in changes of serum protein fingerprinting in PLC patients of different Chinese medical syndromes before and after interventional treatment.</p>

The expression of CD95 and activated antigens in peripheral blood lymphocyte subsets in patients with hand foot and mouth disease (HFMD) / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the expression of CD95 and special marker for activation of peripheral blood lymphocytes in patients with hand foot and mouth disease (HFMD) and its significance.</p><p><b>METHODS</b>Immunofluorescent two-color flow cytometry was used to study the expression of CD95 and HLA-DR on lymphocytes in 58 patients with HFMD and 34 normal controls.</p><p><b>RESULTS</b>Expression of CD3+ T cells was significantly lower in patients (63.82 +/- 7.74)% than that in controls (P < 0.001), meanwhile the expression of CD4+ T cells was (34.29 +/- 7.33)%, significantly lower than that of the controls (P < 0.005). The percentage of lymphocytes expressing HLA-DR in patients was (23.77 +/- 5.78)%, significantly higher than that of the controls (P < 0.005). Significant difference was observed in the expression of HLA- DR on CD8+ T cells in patients (1.34 +/- 1.12)% as compared with controls (P < 0.005). No significant difference in the expression of CD95 on lymphocytes was observed between patients and the controls (P > 0.05).</p><p><b>CONCLUSION</b>The findings support that cellular immunodeficiency exists in patients and that lymphocytes were abnormally activated in the patients. The activation of peripheral blood T lymphocytes in patients mainly involves CD8 subset and it may play an important role in the immune response to antiviral infection.</p>

Study on the changes of the T-lymphocyte subsets and their significance in influenza A (H1N1) virus patients / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study the changes of T-Lymphocyte and activated T-Lymphocyte subsets in influenza A (H1N1) virus patients.</p><p><b>METHODS</b>The percentages of the subsets of Lymphocyte were detected by flow cytometry in influenza A (H1N1) virus patients (n = 144) and normal controls (n = 41). Furthermore, the subsets of T-Lymphocyte and activated T-lymphocyte were analyzed in 83 among those patients before and after treatment.</p><p><b>RESULTS</b>Compared with the control group, the counts of Lymphocyte in patients with influenza A (H1N1) virus was significantly discreased, the counts of Lymphocyte in patients with influenza A virus concurrent pneumonia was significantly discreased those of no concurrent pneumonia; Compared with the control group, the percentage of T-lymphocyte in patients with influenza A virus concurrent pneumonia was significantly discreased. The counts and percentage of CD3 and CD8 cells was significantly discreased in patients (n = 83) before treatment; The counts of CD4 cells was significantly discreased before treatment. The percentage of HLA-DR+ CD+, HLA-DR+ CD4+ and HLA-DR+ CD8+ cells was significantly discreased in patients (n = 83) before treatment.</p><p><b>CONCLUSIONS</b>To understand the expression of the T-Lymphocyte and activated T-Lymphocyte subsets in influenza A (H1N1) virus patients may help to evaluate the patients' cellular immune status, but also be a guideline of early diagnosis of Influenza A (H1N1) virus.</p>

Inhibition of Jingzhaotoxin-V on Kv4.3 channel / 生理学报

Kv4.3 channel is present in many mammalian tissues, predominantly in the heart and central nervous system. Its currents are transient, characterized by rapid activation and inactivation. In the hearts of most mammals, it is responsible for repolarization of the action potential of ventricular myocytes and is important in the regulation of the heart rate. Because of its central role in this important physiological process, Kv4.3 channel is a promising target for anti-arrhythmic drug development. Jingzhaotoxin-V (JZTX-V) is a novel peptide neurotoxin isolated from the venom of the spider Chilobrachys jingzhao. Whole-cell patch clamp recording showed that it partly blocked the transient outward potassium channels in dorsal root ganglion neurons of adult rats with an IC(50) value of 52.3 nmol/L. To investigate the effect of JZTX-V on Kv4.3 channel, JZTX-V was synthesized using the solid-phase chemical synthesis and separated by reverse phase high performance liquid chromatography (HPLC). The purity was tested by matrix-assisted laser desorption-ionization time-of-flight mass spectrometry (MOLDI-TOF mass spectrometry). Two-electrode voltage-clamp technique was used to characterize the action of JZTX-V on Kv4.3 channels expressed in Xenopus laevis oocytes. As a result, JZTX-V displayed fast kinetics of inhibition and recovery from inactivation. Furthermore, it could inhibit Kv4.3 channel current in a time- and concentration-dependent manner with an IC(50) value of 425.1 nmol/L. The application of JZTX-V affected the activation and inactivation characteristics of Kv4.3 channel and caused a shift of the current-voltage relationship curve and the steady-state inactivation curve to depolarizing direction by approximately 29 mV and 10 mV, respectively. So we deduced that JZTX-V is a gating modifier toxin of Kv4.3 channel. Present findings should be helpful to develop JZTX-V into a molecular probe and drug candidate targeting to Kv4.3 channel in the myocardium.

Effect of Chinese herbal treatment on Th1- and Th2-type cytokines, progesterone and beta-human chorionic gonadotropin in early pregnant women of threatened abortion / 中国结合医学杂志

<p><b>OBJECTIVE</b>To study the clinical effects of Prescription Zhuyun-III (ZYIII) on early pregnant women diagnosed as threatened abortion, and its mechanism in immunity and endocrine by determining serum Th1- and Th2-type cytokines, progesterone, and beta-human chorionic gonadotropin (beta-HCG).</p><p><b>METHODS</b>The treatment group comprised 30 early pregnant women diagnosed as threatened abortion of deficiency syndrome of Pi, Shen, or both. The control group consisted of 20 normal early pregnant women of similar gestational age. Patients in the treatment group were administered with ZYIII for 4 weeks. Peripheral blood samples were collected pre- and post-treatment from both the treatment and the control groups. Serum Th1-type cytokine [interleukin-2 (IL-2)] and Th2-type cytokine [interleukin-10 (IL-10)] were determined by flow cytometry, and serum progesterone and beta-HCG were determined by ELISA.</p><p><b>RESULTS</b>(1) The treatment was effective in 26 and ineffective in 4 patients of the treatment group. Therefore, the cure percentage was 86.67%. (2) In the treatment group before the treatment, IL-2 was significantly higher, IL-10 tended to be less, and the Th1/Th2 balance shifted toward Th1 compared with those in the control group. (3) After the treatment, IL-2 was decreased, IL-10 was increased, and IL-2/IL-10 was decreased. Both progesterone and beta-HCG were increased. Changes of progesterone were positively correlated with changes of IL-10, whereas changes of beta-HCG were negatively correlated with changes of IL-2.</p><p><b>CONCLUSIONS</b>Our study suggests that ZYIII has an evident function of protecting the fetus, and one of its mechanisms is inhibiting the secretion of Th1 cytokines, promoting the secretion of Th2 cytokines, and recovering the pathological shift of the Th1/Th2 balance. The other possible mechanism is increasing serum progesterone and beta-HCG concentrations. Moreover, there are some correlations between the above two effects.</p>

Effects of bushen tiaochong recipe containing serum on ovarian granulosa cell proliferation, steroidogenesis and associated gene expression in rats / 中国结合医学杂志

<p><b>OBJECTIVE</b>To observe the effect of bushen tiaochong recipe (BSTCR) on rats' ovarian granulosa cell (GC) proliferation, steroidogenesis and follicle-stimulating hormone receptor (FSHR), and insulin-like growth factor-1 (IGF-1) mRNA expression using serum pharmacological method.</p><p><b>METHODS</b>Rats' GCs were incubated with 10% blank serum (as negative control group), follicle-stimulating hormone (FSH)-containing serum (S-FSH, as positive control group), or BSTCR (in different dosages) containing serum (S-BSTCR, as the BSTCR groups) for 48 h. 3H-TdR incorporation was then performed; DNA was measured to analyze the distribution of GCs in the cell cycle and their proliferation index (PI) using a flow cytometer; estradiol (E2) and progesterone (P) content in the culture fluid were examined by radioimmunoassay; and levels of FSHR and IGF-1 mRNA expression in GCs were measured by real-time RT-PCR.</p><p><b>RESULTS</b>A dose-dependent increase of 3H-TdR incorporation in GC was shown in the BSTCR groups. Cells in G0/G1 phase had markedly less, while those in S phase had a significantly higher increase in the BSTCR groups compared with the negative control group. A high value of PI was also shown in the BSTCR groups, especially in the high dose group where the influence of cell proliferation was stronger than that in the positive control group. The levels of E2 and P in the BSTCR groups of all dosages were significantly higher than those in the negative control group, and did not show any significant difference compared with those in the positive control group. Levels of FSHR and IGF-1 mRNA expression in the BSTCR groups increased in a dose-dependent manner at levels higher than those in the negative control group.</p><p><b>CONCLUSION</b>S-BSTCR can obviously stimulate the proliferation and steroidogenesis of ovarian GCs. It is speculated that BSTCR could play a regulatory action on ovarian function through two different pathways of endocrine and autocrine by promoting FSHR and IGF-1 mRNA expression.</p>

Comparative proteome analysis of human lung squamous cell carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>This study was designed to establish the two-dimensional electrophoresis profiles with high resolution and reproducibility from human lung squamous cell carcinoma tissue and paired tumor-adjacent normal bronchial epithelial tissue, and to identify differential expression of tumor-associated proteins by using proteome analysis.</p><p><b>METHODS</b>Comparative proteome analysis of human lung squamous carcinoma and paired normal bronchial mucosa adjacent to tumors from 20 cases were carried out. Total proteins of the carcinoma tissue and normal bronchial mucosa were separated by means of immobilized pH gradient-based two-dimensional gel electrophoresis (2-DE). The differentially expressed proteins were analyzed and identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS).</p><p><b>RESULTS</b>(1) Seventy-six differentially expressed proteins were screened by analyzing the electrophoretic maps of the 20 carcinoma and control mucosa tissues. (2) Sixty-eight differential proteins were identified by peptide mass fingerprinting (PMF). Some proteins were products of oncogenes and others were involved in the regulation of cell cycle and signal transduction. (3) The expression of three proteins mdm2, c-Jun and EGFR, correlated with lung squamous carcinoma, were detected by immunohistochemical staining and Western blot analysis. The results showed that the expression of mdm2, c-Jun and EGFR were up-regulated in lung squamous carcinomas, whereas down-regulated in control normal mucosa. It was consistent with our proteome analysis results. Those results suggested that those proteins may play roles in the carcinogenesis of lung squamous carcinoma.</p><p><b>CONCLUSION</b>sixty-eight differentially expressed proteins were successfully characterized by comparative proteome analysis. Those results may provide scientific foundation for screening the molecular biomarkers which can be used in diagnosis and treatment of lung squamous carcinoma, as well as to improve patients' prognosis and provide a new clue for carcinogenesis research of lung squamous cell carcinoma.</p>

Solid-phase synthesis and biological characterization of S12A-HNTX-IV and R29A-HNTX-IV: two mutants of hainantoxin-IV / 生物工程学报

Hainantoxin-IV (HNTX-IV) purified from the venom of the spider Selenocosmia hainana is a potent antagonist that acts on tetrodotoxin-sensitive (TrX-S) sodium channels. It is a 35-residue polypeptide and includes three disulfide bridges. In order to investigate the structure-function relationship of HNTX-IV, two mutants (S12A-HNTX-IV and R29A-HNTX-IV) of HNTX-TV in which Ser12 and Arg29 were replaced by Ala respectively, were synthesized by solid-phase Fmoc chemistry, followed by oxidative refolding of purified peptides under the optimal conditions. The synthetic mutants were analyzed by MALDI-TOF mass spectrometry, nuclear magnetic resonance spectroscopy (NMR) and electrophysiological experiments for molecular weight, conformation and physiological activity, respectively. The results show that the mutants and native HNTX-IV (nHNTX-IV) have almost identical three-dimensional structures. The bioactivity level of S12A-HNTX-IV is also about the same as that of nHNTX-IV, suggesting that Ser12 does not play any important role for the bioactivity of this toxin. The bioactivity of R29A-HNTX-IV is reduced by at last 155 times, indicating that Arg29 is a key residue relative to the bioactivity of HNTX-IV. It is presumed that the decrease in activity of R29A-HNTX-IV is due to the changes of the property in the binding site rather than the change in the basic conformation of the molecule.