Advances on nonviral vectors of CRISPR/Cas9 system for genome editing / 药学学报

Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR associated protein system exerts genome editing effect through cleaving DNA double strands using RNA-guided endonuclease. Double-strand breaks were repaired via homology directed repair (HDR) or nonhomologous end joining (NHEJ), accompanied by insertions, deletions or replacements into the genome. As a powerful tool, CRISPR/Cas system has provided tremendous convenience for basic researches and may pave the path to treat genetic diseases and cancers. Genome editing could be achieved only when both CRISPR RNA and Cas protein are delivered into nucleus of target cell. Compared with physical and viral delivery, nonviral delivery of CRISPR/Cas system possesses unique advantages in terms of safety, loading capacity and preparation. Hence, many researchers have devoted themselves to the development of nonviral vectors with high delivery efficiency which is important for the application and translation of the promising technology. Advances on cationic liposomes, lipid like nanoparticles, cationic polymers, AuNPs, vesicles, polypeptides, proteins and so on have been made. We will give a brief introduction to the mechanism of CRISPR/Cas9, problems faced by nonviral delivery of CRISPR/Cas9 system in forms of plasmid, mRNA and protein; examples of non-viral vectors, hoping to give some hints on design of safe and efficient nonviral vectors for genome editing.

Integrated Pharmaceutical Care on Patients with Coronary Heart Diseaseafter First Percutaneous Coronary Intervention / 中国药学杂志

OBJECTIVE: To explore the effect of integrated pharmaceutical intervention on medication compliance, risk factors control, and 6 months and 2 years prognosis in patients with coronary artery disease(CAD) after first percutaneous coronary intervention (PCI) in china. METHODS: 200 cases of patients diagnosed with CAD and underwent PCI successfully firstly in Zhoupu hospital during January 2015 to December were enrolled. The control group was given routine treatment and nursing care; pharmacist intervention group, on this basis, was given pharmaceutical intervention by pharmacists, including optimizing medication program, formulating pharmaceutical care plan, medication education, health education and regular pharmacists outpatient follow-up. The two groups were followed up for two years. Comparison of the Readmission(Readm) rate, incidence rate of major adverse cardiac events (MACEs), Readm expenses, awareness rate of commonly used two-grade preventive drugs for CAD, medication compliance and emotion management after 6 months and two years in the two groups. The control of risk factors in the two groups after 2 years were compared. RESULTS: After six months, the Readm rate, the incidence of MACEs and Readm expenses in pharmacist intervention group were lower than those in the control group, and the medication compliance was slightly higher than the control group (94.9% vs 87.4%); two years later, the standard rate of risk factors in pharmacist intervention group was significantly better than the control group(smoking3.1% vs. 12.6%, P=0.012; blood glucose standard rate79.6% vs. 62.1%, P=0.007; blood pressure standard rate74.5% vs. 53.7%, P=0.003; blood lipid standard rate84.7% vs. 57.9%, P<0.001); and the Readm rate, the incidence of MACEs and the cost of Readm were significantly lower than those of the control group after six months and two years. CONCLUSION: Pharmacists′ active pharmaceutical intervention on patients after PCI can effectively improve the medication compliance of the patients, reduce the postoperative risk factors and the incidence of MACEs, reduce medical expenses, improve the clinical prognosis, which were more significant when the follow-up time was longer.

Spreading of extended-spectrum β-lactamase-producing Escherichia coli ST131 and Klebsiella pneumoniae ST11 in patients with pneumonia: a molecular epidemiological study / 中华医学杂志(英文版)

Background@#Extended-spectrum β-lactamase (ESBL)-producing Escherichia coli (E. coli) and Klebsiella pneumoniae (K. pneumoniae) are the important pathogens causing pneumonia. This study aimed to investigate the clinical characteristics and molecular epidemiology of ESBL-producing E. coli and K. pneumoniae causing pneumonia at a large teaching hospital in China.@*Methods@#We collected patient’s clinical data and ESBL-producing E. coli and K. pneumoniae strains causing pneumonia (from December 2015 to June 2016) at a hospital in Wuhan. The susceptibilities, multi-locus sequence typing, homologous analysis, ESBL genes by polymerase chain reaction and sequencing were determined.@*Results@#A total of 59 ESBL-producing strains (31 E. coli and 28 K. pneumoniae) isolated from patients with pneumonia were analyzed. The majority of strains were isolated from patients were with hospital-acquired pneumonia (37/59, 62.7%), followed by community-acquired pneumonia (13/59, 22.0%), and ventilator-related pneumonia (9/59, 15.3%). The E. coli ST131 (9 isolates, 29.0%) and K. pneumoniae ST11 (5 isolates, 17.9%) were the predominant sub-types. The most prevalent ESBL gene was CTX-M-14, followed by SHV-77, CTX-M-3, SHV-11, and CTX-M-27. At least 33 (55.9%) of the ESBL-producing strains carried two or more ESBL genes. The ISEcp1 and IS26 were found upstream of all blaCTX-M (CTX-Ms) and of most blaSHV (SHVs) (57.6%), respectively. Moreover, three ESBL-producing K. pneumoniae ST11 strains which were resistant to carbapenems carried the blaNDM-1 and blaKPC-2, two of which also bearing blaOXA-48 were resistant to all antibiotics (including Tigecycline).@*Conclusions@#Hospital-acquired pneumonia is more likely correlated with ESBL-producing E. coli and K. pneumoniae. ESBL-producing E. coli ST131 and multi-drug resistance ESBL-producing, as well as New Delhi metallo-β-lactamase-1 (NDM-1) and Klebsiella pneumoniae carbapenemases-2 (KPC-2) bearing K. pneumoniae ST11 are spreading in patients with pneumonia in hospital.

Spreading of extended-spectrum β-lactamase-producing Escherichia coli ST131 and Klebsiella pneumoniae ST11 in patients with pneumonia: a molecular epidemiological study / 中华医学杂志(英文版)

BACKGROUND@#Extended-spectrum β-lactamase (ESBL)-producing Escherichia coli (E. coli) and Klebsiella pneumoniae (K. pneumoniae) are the important pathogens causing pneumonia. This study aimed to investigate the clinical characteristics and molecular epidemiology of ESBL-producing E. coli and K. pneumoniae causing pneumonia at a large teaching hospital in China.@*METHODS@#We collected patient's clinical data and ESBL-producing E. coli and K. pneumoniae strains causing pneumonia (from December 2015 to June 2016) at a hospital in Wuhan. The susceptibilities, multi-locus sequence typing, homologous analysis, ESBL genes by polymerase chain reaction and sequencing were determined.@*RESULTS@#A total of 59 ESBL-producing strains (31 E. coli and 28 K. pneumoniae) isolated from patients with pneumonia were analyzed. The majority of strains were isolated from patients were with hospital-acquired pneumonia (37/59, 62.7%), followed by community-acquired pneumonia (13/59, 22.0%), and ventilator-related pneumonia (9/59, 15.3%). The E. coli ST131 (9 isolates, 29.0%) and K. pneumoniae ST11 (5 isolates, 17.9%) were the predominant sub-types. The most prevalent ESBL gene was CTX-M-14, followed by SHV-77, CTX-M-3, SHV-11, and CTX-M-27. At least 33 (55.9%) of the ESBL-producing strains carried two or more ESBL genes. The ISEcp1 and IS26 were found upstream of all blaCTX-M (CTX-Ms) and of most blaSHV (SHVs) (57.6%), respectively. Moreover, three ESBL-producing K. pneumoniae ST11 strains which were resistant to carbapenems carried the blaNDM-1 and blaKPC-2, two of which also bearing blaOXA-48 were resistant to all antibiotics (including Tigecycline).@*CONCLUSIONS@#Hospital-acquired pneumonia is more likely correlated with ESBL-producing E. coli and K. pneumoniae. ESBL-producing E. coli ST131 and multi-drug resistance ESBL-producing, as well as New Delhi metallo-β-lactamase-1 (NDM-1) and Klebsiella pneumoniae carbapenemases-2 (KPC-2) bearing K. pneumoniae ST11 are spreading in patients with pneumonia in hospital.

Anti-inflammatory effects of compound Cervi Cornu Degelatinatum extract on adjuvant arthritic rats / 中成药

AIM To study the anti-inflammatory effects of compound Cervi Cornu Degelatinatum extract on adjuvant arthritic rats.METHODS The rats,except for those assigned into a blank group,were induced to be the adjuvant arthritis models by Freund's complete adjuvant and randomly divided into model group,Prednisone Acetate Tablets group,Tripterygium wilfordii polyglycoside tablets group,and compound Cervi Cornu Degelatinatum extract groups (1 000 mg/kg high-dose group,500 mg/kg medium-dose group,and 250 mg/kg low-dose group) to observe changes in body weight,mental state,and extent of joints injury.Post-treatment levels of TNF-α,IL-6,IL-10 and PGE-2 were determined by ELISA,and the pathological changes of ankle joints were assessed by HE staining.RESULTS Significantly lower body weight and IL-10 level,markedly higher levels of TNF-α,IL-6,PGE-2,and joints injury in the model group than those in the blank group were observed (P ＜0.01).Such indices also revealed the significant superiority in the high-dose,medium-dose groups of compound Cervi Cornu Degelatinatum extract and the positive control group as compared with the model group.CONCLUSION Compound Cervi Cornu Degelatinatum extract highlights the rheumatoid arthritis management through proinflammatory cytokines secretion reduction,the antiinflammatory factors improvement,and the inflammatory reaction and tissue damage alleviation.

In vitro targeting efficiency evaluation of reduction-responsive co-loaded doxorubicin/siRNA nanoparticles / 药学学报

In this study a reduction-responsive nanoparticles (NPs) modified with hyaluronic acid (HA) was prepared for the co-delivery of doxorubicin (DOX) and siRNA and then evaluated as a lung cancer targeting delivery system in vitro. The amphiphilic polymer of poly-L-lysine-lipoic acid (PLA) based on poly-L-lysine (PLL) with lipoic acid (LA) was synthesized via amidation reaction and characterized by 1H NMR. The DOX loaded PLA NPs were prepared via dialysis method, and siRNA was loaded via electrostatic attraction to prepare the co-delivery NPs system (PLA/DOX-siRNA-NPs). Then PLA/DOX-siRNA-NPs were coated with HA to obtain HA-PLA/DOX-siRNA-NPs. The tumor microenvironment-responsive properties under different pH or reduction condition of HA-PLA/DOX-siRNA-NPs were evaluated by investigating the particle size and zeta potential. Cellular uptake of HA-PLA/DOX-siRNAFAM-NPs by A549 cells and endosomal escape of siRNA were studied using confocal laser scanning microscope (CLSM). 1H NMR spectrum demonstrated that PLA was successfully synthesized with LA grafting rate of 25.1%. The encapsulation efficiency (EE) and drug loading (DL) of HA-PLA/DOX-NPs was (86.93±8.91)% and (4.17±0.68)%, respectively, and siRNA was loaded at an N/P of 6:1 in carrier. HA-PLA/DOX-siRNA-NPs exhibited a suitable size of (167.3±9.9) nm and negative charge of (-15.5±1.4) mV with the optimal ratio of PLA and HA of 1:3. Additionally, the zeta potential of HA-PLA/DOX-siRNA-NPs significantly increased with charge reversal from negative to positive after the treatment with HAase, and the particle size of HA-PLA/DOX-siRNA-NPs changed significantly under the condition of 10 mmol·L-1 glutathione (GSH). The release profiles in vitro demonstrated that HA-PLA/DOX-NPs exhibited a maintained release behavior at pH 7.4 and the adding of GSH (10 mmol·L-1) led to rapid release of DOX from NPs. In vitro cellular uptake and subcellular distribution study demonstrated that themodification of HA enhanced the affinity of NPs to A549 cells and targeting ability, and the cellular uptake of HA-PLA/DOX-siRNAFAM-NPs significantly increased after the treatment with HAase. It was observed that HA-PLA/DOX-siRNAFAM-NPs could escape from endo-lysosomes followed by sharp payloads release to their relative targets. All these results demonstrated that the co-loaded NPs have a high entrapment efficiency of DOX and siRNA. And they also exhibited an active tumor targeting efficiency and tumor microenvironment-responsive properties, which were beneficial to cellular uptake and intracellular release of DOX and siRNA. In conclusion, these reduction-responsive NPs modified with HA have great potential as co-delivery systems for antitumor agents and siRNA.

Clinical observation on treatment of type 2 cardiac and kidney syndrome by combination of traditional Chinese and Western medicines / 中国中药杂志

Clinical observation on treatment of type 2 cardiac and kidney syndrome by combination of traditional Chinese and Western medicine. The patients were divided into two groups： the simple Western medicine treatment group (control group) and the traditional Chinese medicine and Western medicine treatment group (treatment group). The patients in the two groups were treated with conventional western medicine.The treatment group was given based on Buxin Yishen decoction, a total of three courses of treatment to observe the two groups of patients before and after treatment of total efficacy, cardiac function indicators, changes in renal function indicators. The total efficacy of the treatment group and the control group were 91.80% and 72.41%, respectively. There were significant differences between the two groups (P<0.01). The cardiac function indexes and renal function indexes of the treatment group and the control group before and after treatment (P<0.01). Compared with the two groups, the left ventricular function, Hematuria natriuretic peptide, serum creatinine, urea nitrogen, cystatin-C were improved, and the treatment group (P<0.05~0.01). The results showed that the combination of traditional Chinese and Western medicine treatment can improve the clinical efficacy of type 2 heart and kidney syndrome, significantly improve heart and kidney function, better than conventional Western medicine treatment, and has good safety.

pH-sensitive micelles loaded paclitaxel using carboxymethyl chitosan-palmitic acid mediated by cRGD / 药学学报

cRGD-carboxymethyl chitosan-palmitic acid (cRGD-CMCh-PA) was synthesized and a pHsensitive paclitaxel-loaded cRGD-CMCh-PA micelles (PTX-cRGD-CMCh-PA) was prepared with the film dispersion method; related substances were characterized by FT-IR and 1H NMR. PTX-cRGD-CMCh-PA micelles were studied with the particle size distribution, zeta potential, morphology and release behavior in vitro was investigated by the method of equilibrium dialysis. In vitro cytotoxicity of different formulations on A549 cells was tested by MTT assay. The uptake process of micelles was explored using confocal microscopy and a live cell station was used to observe the dynamic phagocytosis. The subcutaneous and orthotropic tumor models were built to study the distribution of DiR-labeled micelles by near-infrared fluorescence (NIR) imaging system. The FT-IR spectra and 1H NMR spectra confirmed the successful conjugation of cRGD-CMCh-PA polymer and the degree of carboxymethyl and the palmitic acid grafted on chitosan were 45.0% and 15.0%. PTX-cRGD-CMCh-PA micelles were prepared with particle size of (162.9±1.5) nm, zeta potential of +26.3 mV and encapsulation efficiency and the drug loading of 99.67% and 28.5%, respectively. The micelles released slowly in pH 7.4 whose release curves were accorded with the Higuchi equation; they had an initial burst effect in second hours and showed a pH sensitive release behavior in pH 5.3. The IC50 of PXT-CMCh-PA and PTX-cRGD-CMCh-PA were 2.077 μg·mL-1 and 0.876 μg·mL-1, respectively. The cells uptake process of micelles in A549 cells revealed that the micelles were mainly co-located with lysosome and PTX-cRGD-CMCh-PA showed much better targeting effect. The NIR fluorescence imaging results showed that the micelles had a good targeting effect on both subcutaneous and orthotropic tumors. In this study, a novel copolymer cRGDCMCh-PA was synthesized with a sustained and pH-dependent drug release activity which would potentially become a new carrier for hydrophobic drugs.

Research progress on the role of transcription factor Nrf 2 in diabetic nephropathy / 中国糖尿病杂志

Oxidative stress and inflammation are two main factors in the progress of diabetic nephropathy(DN) and its complications. NF‐E2 p45‐related factor (Nrf2) is a crucial transcriptional factor which manipulates downstream genes that encode some antioxidant enzymes and phase Ⅱ detoxifying enzymes ,to maintain the redox homeostasis and cellular detoxification response. Therefore ,more and more researchers are focusing on the role of Nrf2 in DN. In this review ,the detailed role of Nrf2 in DN will be discussed. Hopefully ,our work can epitomize recent research progress and provide novel clues for diabetic nephropathy prevention and treatment.

Effects of p38 MAPK signaling pathway and aldose reductase on transforming growth factor-β1 induced expression of fibronectin in cultured human mesangial cells / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the effects of p38 MAPK signaling pathway and aldose reductase (AR) on the transforming growth factor (TGF)-β1-induced expression of fibronectin (FN).</p><p><b>METHODS</b>Human mesangial cells (HMCs) were cultured, and transfected with pCDNA3-AR. AR gene silencing was induced by small interfering RNA (siRNA). AR expression in HMCs was examined by immunofluorescence analysis. RT-PCR and real-time PCR were performed to detect the mRNA expression of AR in the HMCs and Western blotting was used to detect the protein expression of AR, FN and p38. AR inhibitors (ARIs), Sorbinil and Zopolrestat were added and co-incubated, followed by addition of TGF-β1. Western blotting was used to document protein expression of FN and p38 mitogen-activated protein kinases (p38 MAPKs) in the HMCs.</p><p><b>RESULTS</b>Immunofluorescence analysis showed a stronger expression of AR in HMCs transfected with AR than that of normal HMCs and HMCs transfected with blank vector. In comparison with normal HMCs and those transfected with blank vector, HMCs transfected with AR showed stronger protein expression of FN (P<0.05). After incubation of ARIs, protein expression of FN decreased in HMCs transfected with AR (P<0.05). After stimulation of TGF-β1, FN protein expression increased in both normal HMCs and those transfected with AR (P<0.05). After preincubation with ARI, FN protein expression in HMCs transfected decreased significantly (P<0.05). After stimulation of TGF-β1, naïve HMCs showed increased expression of phosphor-p38. In contrast, HMCs preincubated with ARIs showed reduced expression of phosphor-p38, and HMCs transfected with AR showed increased expression of phosphor-p38 (P<0.05).</p><p><b>CONCLUSIONS</b>AR regulates the expression of FN through the stimulation of TGF-β1, which may involve the activation of p38-MAPK signaling pathway. AR may play a role in the pathogenesis of glomerulosclerosis.</p>

Dioscin-induced apoptosis of human LNCaP prostate carcinoma cells through activation of caspase-3 and modulation of Bcl-2 protein family / 华中科技大学学报（医学）（英德文版）

Dioscin is a natural steroid saponin derived from several plants, showing potent anti-cancer effect against a variety of tumor cell lines. In the present study, we investigated the anti-cancer activity of dioscin against human LNCaP cells, and evaluated the possible mechanism involved in its antineoplastic action. It was found that dioscin (1, 2 and 4 μmol/L) could significantly inhibit the viability of LNCaP cells in a time- and concentration-dependent manner. Flow cytometry revealed that the apoptosis rate was increased after treatment of LNCaP cells with dioscin for 24 h, indicating that apoptosis was an important mechanism by which dioscin inhibited cancer. Western blotting was employed to detect the expression of caspase-3, Bcl-2 and Bax in LNCaP cells. The expression of cleaved caspase-3 was significantly increased, and meanwhile procaspase-3 was markedly decreased. The expression of anti-apoptotic protein Bcl-2 was down-regulated, whereas the pro-apoptotic protein Bax was up-regulated. Moreover, the Bcl-2/Bax ratio was drastically decreased. These results suggested that dioscin possessed potential anti-tumor activity in human LNCaP cells through the apoptosis pathway, which might be associated with caspase-3 and Bcl-2 protein family.

Dioscin-induced apoptosis of human LNCaP prostate carcinoma cells through activation of caspase-3 and modulation of Bcl-2 protein family / 华中科技大学学报（医学）（英德文版）

Dioscin is a natural steroid saponin derived from several plants, showing potent anti-cancer effect against a variety of tumor cell lines. In the present study, we investigated the anti-cancer activity of dioscin against human LNCaP cells, and evaluated the possible mechanism involved in its antineoplastic action. It was found that dioscin (1, 2 and 4 μmol/L) could significantly inhibit the viability of LNCaP cells in a time- and concentration-dependent manner. Flow cytometry revealed that the apoptosis rate was increased after treatment of LNCaP cells with dioscin for 24 h, indicating that apoptosis was an important mechanism by which dioscin inhibited cancer. Western blotting was employed to detect the expression of caspase-3, Bcl-2 and Bax in LNCaP cells. The expression of cleaved caspase-3 was significantly increased, and meanwhile procaspase-3 was markedly decreased. The expression of anti-apoptotic protein Bcl-2 was down-regulated, whereas the pro-apoptotic protein Bax was up-regulated. Moreover, the Bcl-2/Bax ratio was drastically decreased. These results suggested that dioscin possessed potential anti-tumor activity in human LNCaP cells through the apoptosis pathway, which might be associated with caspase-3 and Bcl-2 protein family.

Pharmacodynamic study of racemic TJ0711 on renal hypertensive rats after long-term administration / 药学学报

The study is to observe the effect of racemic TJ0711 on blood pressure and heart rate as well as protection of cardiovascular system of renal hypertensive rats after long-term administration. The renal hypertensive models were established by the two-kidney, one-clip (2K1C) method in Wistar rats. Four weeks later, assigned the rats whose SBP had increased at least 4 kPa randomly into 5 groups: racemic TJ0711 10, 20 and 40 mg x kg(-1) groups, carvedilol control group, model group and sham group (n=10), ig administration once daily. The changes of BP (blood press) and HR (heart rate) before and after administration were measured by tail-cuff method weekly. Plasma samples of all animals were taken in 6-8 weeks, and plasma MDA as well as renin, angiotensin II (Ang II) and endothelin-1 (ET-1) levels were measured. Left ventricle was cut off after 9 weeks, and left ventricular weight index (LVWI) and hydroxyproline were measured. The significant decrease of the BP of TJ0711 40 mg x kg(-1) group was observed after TJ0711 ig administration for 4 weeks, and this effect remained till the end of the study. In 8th week, the systolic blood pressure values were: TJ0711 40 mg x kg(-1) group 18.93 +/- 1.82 kPa (vs 21.30 +/- 2.30 kPa, P < 0.05); 20 mg x kg(-1) group 20.68 +/- 3.29 kPa (vs 22.19 +/- 2.88 kPa). The plasma MDA level of all treated groups was significantly lower than that of model group, so were the plasma renin, Ang II and ET-1 levels (P < 0.05). LVWI and hydroxyproline content of myocardial tissue decreased to some extent, but was not significant as compared with that of model group. The study showed that TJ0711 repeated dosing could reduce BP level beginning from drug administration; besides block adrenal alpha and beta receptors to play an antihypertensive role. The sustained antihypertensive effect also related to reduce plasma vasoconstrictor substances and oxidation product MDA. These effects benefited cardiovascular protection.

Pentoxifylline attenuates cigarette smoke-induced overexpression of CXCR3 and IP-10 in mice / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Cigarette smoke-induced emphysema is associated with overexpression of the chemokine receptor CXCR3 and its ligands. Previously, we have demonstrated that pentoxifylline (PTX) alleviated cigarette smoke-induced emphysema. The aim of this study was to determine if the overexpression of CXCR3 and its ligand interferon-inducible protein-10 (IP-10) that was elicited by smoke exposure were attenuated by PTX.</p><p><b>METHODS</b>(1) The study in vitro: a given number of RAW264.7 macrophages with decreasing concentrations of PTX in the culture medium were challenged with cigarette smoke extract (CSE); (2) The study in vivo: male BALB/c mice were randomized into four groups, i.e., sham-smoke, smoke only, smoke with 2 mg/kg PTX, and smoke with 10 mg/kg PTX. The smoke exposure time was 90 minutes once a day, 6 days a week for 16 weeks. PTX was given intraperitoneally before each episode of smoke exposure. Interferon (IFN)-γ and IP-10 in broncho-alveolar lavage fluid (BALF) and in culture medium were measured by enzyme-linked immunosorbent assay (ELISA). IP-10 mRNA in lung tissue was assessed by RT-PCR. CXCR3 positive cells in lung sections were visualized by immunochemistry staining.</p><p><b>RESULTS</b>Up-regulation of IFN-γ and IP-10 in the culture medium of macrophages elicited by CSE was inhibited by PTX in a dose-dependent manner. Chronic cigarette smoke exposure led to overexpression of IFN-γ and IP-10 in BALF, upregulation of IP-10 mRNA and increased infiltration of CXCR3(+) cells into lung parenchyma. Administration of PTX decreased the level of IFN-γ from (6.26 ± 1.38) ng/ml to (4.43 ± 0.66) ng/ml by low dose PTX or to (1.74 ± 0.28) ng/ml by high dose PTX. IP-10 was reduced from (10.35 ± 1.49) ng/ml to (8.19 ± 0.79) ng/ml by low dose PTX or to (7.51 ± 0.60) ng/ml by high dose PTX. The expression of IP-10 mRNA was also down-regulated (P < 0.05). But only with a high dose of PTX was the ratio of CXCR3(+) cells decreased; 15.2 ± 7.3 vs. 10.4 ± 1.8 (P < 0.05).</p><p><b>CONCLUSION</b>PTX attenuates cigarette smoke-induced overexpression of chemokine receptor CXCR3 and its ligand IP-10, which is relevant to its inhibitory effect on pulmonary emphysema.</p>

Genetic characteristics of hantaviruses carried by Microtus maximowixzii in Yakeshi of Inner Mongolia, China / 中华流行病学杂志

Objective To analyze the viral genetic characteristics of hantaviruses carried by Microtus maximowixzii in Yakeshi of Inner Mongolia Autonomous Region and its relationship with Hantaan virus (HTNV) and Seoul virus (SEOV) viruses as well as to identify the natural host of Khabarovsk virus (KHAV).Methods HV specific RNAs were detected by RT-PCR.Complete S and M segment were amplified from the RNA-positive samples.Phylogenetic analysis were performed to estimate the genetic characterization and the relationship with other hantaviruses.Results Fifty two Microtus maximowixzii voles were captured in Yakeshi areas.Of those voles,hanta-viral RNA was tested positive in 5 samples (9.62％).Complete S and M segments sequences were obtained from 5 and 2 lung samples,respectively.The complete S segment was consisted of 1848 to 1861 bp,and the M segment consisted of 3662 bp.These viruses were closely related to each other with 92.5％-96.4％ for the S segment sequences and 88.9％-95.4％ for the M segment sequences.They shared a higher identity with KHAV found previously in Yakeshi and KHAV of Russia.However,they were obviously different from the other hantavirus species.The 5 strains had the consistent secondary structure of nucleocapsid protein (NP) and glycoprotein (GP).When further comparing their secondary structures with those of HTNV and SEOV,our results indicated that there were no obvious differences in NP between KHAV and both HNTV,SEOV but with obvious difference in GP.Based on the S and M segment sequences,phylogenetic analyses revealed that these 5 strains clustered together with KHAV and formed a distinct lineage.Furthermore,all known KHAV strains could be divided into two small branches with a nucleotide divergence more than 5.3％.Conclusion Our research data revealed that KHAV was highly endemic among Microtus maximowixzii in Yakeshi area which supported the notion that Microtus maximowixzii had been the natural host of KHAV in the area.

Elevated nocturnal and morning blood pressure in patients with obstructive sleep apnea syndrome / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The nocturnal nondipping and elevated morning blood pressure (BP) in patients with obstructive sleep apnea syndrome (OSAS) have not yet been well investigated in Chinese patients. This study aimed to describe the BP profile, and to elucidate the relationships between daytime BP and nighttime BP, and between evening BP and morning BP in patients with OSAS.</p><p><b>METHODS</b>Twenty teaching hospital sleep centers in China were organized by the Chinese Medical Association to participate in this study and 2297 patients were recruited between January 2004 and April 2006. BP assessments were made at four time points (daytime, evening, nighttime and morning) and polysomnography (PSG) was performed and subjects were classified into four groups by their apnea-hypopnea index (AHI): control, n = 213 with AHI < 5; mild, n = 420 with AHI ≥ 5 and < 15; moderate, n = 460 with AHI ≥ 15 and < 30; and severe, n = 1204 with AHI ≥ 30. SPSS 11.5 software package was used for statistical analysis and figure drawing.</p><p><b>RESULTS</b>All the average daytime, nighttime, evening and morning BPs were positively correlated with AHI and negatively correlated with nadir nocturnal oxygen saturation. The ratios of nighttime/daytime and morning/evening average BP were positively correlated with AHI. The ratio of nighttime/daytime systolic BP became a "reversed BP dipping" pattern until the classification reached severe, while the ratio of nighttime/daytime diastolic BP became reversed at moderate. Similarly, the ratio of morning/evening diastolic BP becomes reversed even at mild.</p><p><b>CONCLUSIONS</b>OSAS may result in higher BP levels at all four time points. The ratios of nighttime/daytime and morning/evening BP increase with increased AHI. The increasing of diastolic BP, which is inclined to rise more quickly, is not parallel with increasing systolic BP.</p>

Improved osteoblast proliferation, differentiation and mineralization on nanophase Ti6Al4V / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Previous studies have demonstrated increased functions of osteoblasts on nanophase materials compared to conventional ceramics or composites. Nanophase materials are unique materials that simulate dimensions of constituent components of bone as they possess particle or grain sizes less than 100 nm. However, to date, interactions of osteoblasts on nanophase materials compared to conventional metals remain to be elucidated. The objective of the present in vitro study was to synthesize nanophase metals (Ti6Al4V), characterize, and evaluate osteoblast functions on Ti6Al4V. Such metals in conventional form are widely used in orthopedic applications.</p><p><b>METHODS</b>In this work, nanophase Ti6Al4V surfaces were processed by the severe plastic deformation (SPD) principle and used to investigate osteoblast long-term functions. Primary cultured osteoblasts from neonatal rat calvaria were cultured on both nanophase and conventional Ti6Al4V substrates. Cell proliferation, total protein content, and alkaline phosphatase (ALP) activity were evaluated after 1, 3, 7, 10 and 14 days. Calcium deposition, gene expression of type I collagen (Col-I), osteocalcin (OC), osteopontin (OP) and the production of insulin-like growth factor-I (IGF-I) and transforming growth factor-beta 1 (TGF-β1) were also investigated after 14 days of culture.</p><p><b>RESULTS</b>Functions of osteoblasts including proliferation, synthesis of protein, and ALP activity were improved on the nanophase compared to the conventional Ti6Al4V. The expression of Col-I, OC and OP mRNA was also increased on nanophase Ti6Al4V after 14 days of culture. Calcium deposition was the same; the average number of the calcified nodules on the two Ti6Al4V surfaces was similar after 14 days of culture; however, highly significant size calcified nodules on the nanophase Ti6Al4V was observed. Of the growth factors examined, only TGF-β1 showed a difference in production on the nanophase surface.</p><p><b>CONCLUSION</b>Nanophase Ti6Al4V surfaces improve proliferation, differentiation and mineralization of osteoblast cells and should be further considered for orthopedic implant applications.</p>

Investigation of children's intelligence quotient and dental fluorosis in drinking water-type of endemic fluorosis area in Pucheng county Shaanxi province before and after drinking water change / 中国地方病学杂志

Objective To further understand the effect of anti-fluoride water on intellectual development of children and dental fluorosis in drinking water-type of endemic fluorosis area Pucheng county Shaanxi province.Methods Two hundred 8 to 12 years old children were randomly selected in endemic fluorosis area with changed water or unchanged water, in 2009 in Pucheng county Shaanxi province. Intelligence quotient(IQ) of children was measured by Raven's test. Children's dental fluorosis was examined by Dean's classification scheme. Results The rates of child dental fluorosis in changed water and unchanged water endemic fluorosis areas were 28.50%(57/200), 87.88%(203/231) , respectively, the difference was statistically significant(x2 = 159.19, P＜ 0.01),while dental fluorosis indices were 0.57,1.97, was marginal, moderate epidemic. Children with IQ above 90 was accounted for 89.5% (179/200), 91.5% (183/200) in changed water and unchanged water endemic fluorosis area,respectively, the difference was not significant (x2 = 0.46, P ＞ 0.05). Conclusions Changed water may decrease the incidence of child dental fluorosis, but has no obvious effect on children's IQ development in endemic fluorosis area.

Effect of ERK signalling pathway and aldose reductase on the expression of fibronectin induced by transforming growth factor β1 in human mesangial cells / 中华肾脏病杂志

Objective To study the effect of ERK signalling pathway and aldose reduetase(AR)on the transforming growth factor-β1(TGF-β1)-induced expression of fibronectin (FN)in nondiabetic nephropathy. Methods Human mesangial cells(HMCs)were cultured and transfected with pCDNA3-AR and AR gene silencing with small interfering RNA(siRNA).The AR expression in the HMCs was examined by real-time PCR and Western blotting was used to detect the protein expression of AR and FN. Inhibitors of AR and ERK signalling pathway were co-cultured with HMCs, then TGF-β1 was added and Western blotting was used to analyze the protein expression of FN. Results The expression of AR, FN and p-ERK was up-regulated by TGF-β1. AR was increased by 1.8-fold, FN was increased by 1.9-fold (P＜0.05), and p-ERK was increased by 5.l-fold after stimulation with TGF-β1 (P＜0.01). HMCs transfected with AR showed stronger protein expression of FN, more than 3.6-fold in the protein level of FN was observed in HMCs (P＜0.05). The HMCs of knockdown AR gene by siRNA showed decreased expression of AR,FN and p-ERK, the level of AR mRNA in HMCs transfected with AR siRNA was 10% of the level in untransfected cells or cells transfected with control siRNA (P＜0.01). Transfection with AR siRNA attenuated TGF-β1-induced FN production, more than 70% decrease in the protein level of FN and p-ERK was observed in HMCs with AR-siRNA (P＜0.01). Conclusions AR can regulate the expression of FN with the stimulation of TGF-β1, as AR gene is one of the responsive genes of TGF-β1, which may be associated with the activation of ERK signalling pathways induced by TCF-β1.

Effects of pentoxifylline on Wnt/β-catenin signaling in mice chronically exposed to cigarette smoke / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Previous discovery that long-term administration of pentoxifylline (PTX) to mice chronically exposed to smoke led to the development of pulmonary fibrosis rather than emphysema initiated our curiosity on whether the Wnt/β-catenin pathway, a set of signaling proteins essential to organ development and lung morphogenesis in particular were activated in the pathogenesis of pulmonary fibrosis.</p><p><b>METHODS</b>Male BALB/c mice were randomized into four study groups: Group Sm, smoke exposure and taken regular forage; Group PTX, no smoke but taken PTX-rich forage; Group Sm + PTX, smoke exposure and taken PTX-rich forage; Group control: shamed smoke exposure and taken regular forage. Animals were sacrificed at day 120. Morphometry of the lung sections and the expressions of TGF-β(1), hydroxyproline, β-catenin, cyclin D1, T cell factor 1 (Tcf-1) and lymphoid enhancer factor 1 (Lef-1) mRNA, etc, in the lung homogenate or in situ were qualitatively or quantitatively analyzed.</p><p><b>RESULTS</b>As expected, smoke exposure along with PTX administration for 120 days, lungs of the mice progressed to be a fibrosis-like phenotype, with elevated fibrosis score (3.9 ± 1.1 vs. 1.7 ± 0.6 in Group Sm, P < 0.05). TGF-β(1) (pg/g) (1452.4 ± 465.7 vs. 818.9 ± 202.8 in Group Sm, P < 0.05) and hydroxyproline (mg/g) (5.6 ± 0.6, vs. 2.4 ± 0.1 in Group Sm, P < 0.05) were also consistently increased. The upregulation of β-catenin measured either by counting the cell with positive staining in microscopic field (17.4 ± 7.9 vs. 9.9 ± 2.9 in Group Sm, P < 0.05) or by estimation of the proportion of blue-stained area by Masson's trichrome (11.8 ± 5.6 vs. 4.7 ± 2.4 in Group Sm) in Group SM + PTX was much more noticeable as than those in Group Sm. The expression of β-catenin measured by positive cell counts was correlated to TGF-β(1) concentration in lung tissue (r = 0.758, P < 0.001). PTX per se caused neither fibrosis nor emphysema though expression of β-catenin and downstream gene cyclin D(1) may also be altered by this medication.</p><p><b>CONCLUSIONS</b>PTX mediated transformation of pulmonary emphysema into pulmonary fibrosis under chronic cigarette smoke exposure is associated with upregulation of β-catenin and elevation of TGF-β(1), implying that activation of Wnt/β-catenin signaling may be involved in the pathogenesis of pulmonary fibrosis.</p>