Membrane proteins are the main undertakers of biofilm function, and also the most important target group for innovative drug discovery and research. About 60% of drugs targets are membrane proteins. Due to the obvious aggregation and denaturation tendency of membrane proteins in aqueous solution, it is difficult to simulate the membrane like environment to maintain the correct conformation of membrane proteins in vitro, which results in the slower-growing research on the structure and function of membrane proteins and related ligand drugs than that of water-soluble proteins. Membrane protein stabilization technology is the premise of establishing high specificity, high sensitivity and high throughput drug screening methods for membrane protein ligands, which is of great significance. In this paper, some techniques for stable separation and purification of membrane proteins are reviewed, including detergents, artificial membranes, polymers, lentiviral particles and so on, as well as their specific applications in drug screening.

OBJECTIVETo establish a reliable model for drug screening and therapy by culturing rat femoral head and inducing cartilage degeneration quickly in vitro.

METHODSThe femoral heads from the same SD rats of two-month old were divided into control group and experimental group respectively. They were cultured with DMEM medium plus 10% fetal bovine serum or DMEM medium plus 10% fetal bovine serum plus 50 ng/ml IL-1β for three days. Femoral heads were fixed in 4% paraformaldehyde, decalcified, dehydrated, embedded in paraffin and cut into slices. Specimens were stained with Toluidine blue and Safranine O-Fast Green FCF. The protein expression levels of type II collagen, MMP13, Sox9 and ADAMTS5 were analyzed by immunofluorescence.

RESULTSBoth the Toluidine blue and Safranine O staining were pale in the margin of femoral heads which were stimulated with IL-1β for three days compared to that in control group. The Fast Green FCF staining was positive at the edge of the femoral head in experimental group, which indicated that cartilage became degenerated. The expression levels of both type H collagen and Sox9 were decreased significantly while the expression levels of MMP13 and ADAMTS5 were increased in experimental group.

CONCLUSIONThe model of cartilage degeneration is established by culturing and inducing the degeneration of the femoral heads quickly in vitro.

Animals ; Cartilage Diseases ; genetics ; metabolism ; Collagen Type II ; genetics ; metabolism ; Disease Models, Animal ; Femur Head ; metabolism ; Humans ; In Vitro Techniques ; Interleukin-1beta ; genetics ; metabolism ; Male ; Matrix Metalloproteinase 13 ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; SOX9 Transcription Factor ; genetics ; metabolism

ABC transporters on the intestinal barrier, blood-brain barrier and on tumor cells will affect drug bioavailability, transport across the blood-brain barrier and multidrug resistance. The active ingredients of traditional Chinese medicines can affect the function and expression of ABC transporters. When combined with pharmaceuticals the potential interaction between the two can change the efficacy of the medicines. We review the ABC transporter superfamily and their distribution with regard to their relationship and interactions with traditional Chinese medicine on the intestinal barrier and the blood-brain barrier, as well as their role in tumor multidrug resistance mediated by ABC transporters. We summarize the research progress over the past five years.

Objective: To observe the efficacy of herbal cake-partitioned moxibustion for lumbar disc herniation (LDH) due to kidney deficiency and blood stasis and observe the influence of this method on lumbar functions and inflammatory factors in patients with this condition. Methods: A total of 120 LDH patients who met the inclusion criteria were randomly divided into three groups, including a herbal cake-partitioned moxibustion group, a flour cake-partitioned moxibustion group, and a Western medication group, with 40 patients in each group. The patients in the Western medication group were treated with diflunisal tablets, 0.5 g per dose, 2 doses a day. Those in the herbal cake-partitioned moxibustion group were treated with additional herbal cake-partitioned moxibustion group at Back-Shu Points and Jiaji (EX-B2) Points once a day. Those in the flour cake-partitioned moxibustion group were treated with the same methods as in the herbal cake-partitioned moxibustion group, except that the herbal cake was replaced by a flour cake. All the patients were treated for 10 d. After treatment, the scores of the visual analog scale (VAS) and Japanese Orthopaedic Association (JOA) and the changes of the interleukin (IL)-6, tumor necrosis factor (TNF)-α, and substance P (SP) levels were observed, and the efficacy was evaluated. Results: After treatment, the VAS score and the levels of IL-6, TNF-α, and SP were lower than those before treatment, and the JOA score was higher than that before treatment in the three groups, indicating intra-group statistical significance (P<0.05). The VAS score and the levels of IL-6, TNF-α, and SP of the herbal cake-partitioned moxibustion group were lower than those of the flour cake-partitioned moxibustion group and the Western medication group, while the JOA score of the herbal cake-partitioned moxibustion group was higher than that of the other two groups, indicating inter-group statistical significance (P<0.05). The total effective rate of the herbal cake-partitioned moxibustion group was 92.5％, higher than that of the flour cake-partitioned moxibustion group (80.0％) and the Western medication group (72.5％), indicating inter-group statistical significance (P<0.05). Conclusion: On the basis of Western oral medication, additional herbal cake-partitioned moxibustion can alleviate the pain and improve the lumbar functions in patients with LDH due to kidney deficiency and blood stasis. The efficacy of the integrated method is better than that of either flour cake-partitioned moxibustion or Western medication alone, which may be related to the reduction of serum inflammatory factors.

BACKGROUNDRecently, the number of patients with prostate cancer who needed to be treated with radical prostatectomy increased rapidly in China. There is still a difference between clinical staging and the post-operative final pathologic staging; hence, an excellent tool for accurately predicting the pathologic stages of prostate cancer is needed urgently in clinical practice. The Partin tables are the most popular and widely used tool for predicting the pathologic stages of prostate cancer because of its high accuracy and ease of implementation. The aim of this study was to externally validate the accuracy of the three versions of the Partin tables in predicting the post-operative pathologic stages in Chinese patients with prostate cancer.

METHODSWe retrospectively analyzed the data from 203 patients with prostate cancer who underwent radical prostatectomies between June 2000 and May 2012. The accuracies of the three versions of the Partin tables in predicting the post-operative pathologic stages in Chinese patients with prostate cancer were evaluated using the area under the receiver operating characteristic curve (AUC).

RESULTSUsing the 1997, 2001, and 2007 Partin tables for predicting the current cases, the AUC of organ confinement (OC) was 0.877, 0.788, and 0.726; the AUC of extracapsular extension (ECE) was 0.525, 0.615, and 0.608; the AUC of seminal vesicle invasion (SVI) was 0.875, 0.649, and 0.820; and the AUC of pelvic lymph node invasion (LNI) was 0.808, 0.758, and 0.735 respectively.

CONCLUSIONSThe accuracies of the three versions of Partin tables in predicting OC, SVI, and LNI were good, especially the 2001 Partin table for SVI. In contrast, the accuracy of the three versions of the Partin tables in predicting ECE was fair. The 1997 Partin table was much better than the 2007 table in predicting OC, and the 2001 table in predicting SVI. The 2007 Partin table did not show any advantages.

Aged ; Humans ; Male ; Middle Aged ; Neoplasm Grading ; Neoplasm Staging ; Nomograms ; Prostate-Specific Antigen ; blood ; Prostatectomy ; Prostatic Neoplasms ; pathology ; surgery ; Retrospective Studies

Objective:To methodologically establish the lentivirus granule packaging, concentration and infection against CD34~+ cells from umbilical blood. Methods:The lentivirus system of the 3~(rd) generation was used to produce the virus. Ultrafiltration and ultracentrifugation were employed to concentrate virus. Several treatments were used to improve virus infection including in vitro amplification culture, facilitation of rest cells into cell cycle, promotion of cell adhesion and immobilization during infection, and repeat infection methods. Results:CD34~+ cells were not obviously changed by checking the expression level of CD34 marker on the cell surface after 48 h culture. After two-step concentration, virus titer was increased up to 5.06×10~7/ml, and the infection rate against CD34~+ cells from umbilical blood was increased up to 37.7%.Conclusion:Lentivirus supernatant with over 10~7/ml titer can be obtained using the above methods. Efficient infection against CD34~+ cells from umbilical blood can be achieved.

OBJECTIVETo investigate the clinical manifestations, genotypes, and genetic characteristics of two pedigrees with Kennedy disease.

METHODSThe clinical data of the patients from two Kennedy disease families were collected. The numbers of trinucleotide CAG repeats in exon 1 of the androgen receptor gene were determined by DNA sequencing and repeat fragment analysis.

RESULTSFamily A was composed of 58 individuals in 4 generations. The proband had onset at 39 years old. There were two Kennedy disease patients in family B which included 61 individuals in 5 generations. The two patients had onset at 39 and 41 years old, respectively. All the three patients displayed limbs and bulbar muscular weakness because of the damage of lower motor neurons. They had androgen insensitivity syndrome in common, and showed mild or moderate increase in serum creatine kinase level. The electromyogram showed wild damage in anterior horn of spinal cord. Muscle biopsy displayed neurogenic muscular atrophy. The numbers of the CAG repeat expansion in the androgen receptor gene of the three patients were 49, 48, and 47, respectively. X-linked recessive mode of inheritance was demonstrated by pedigree analysis in the two families.

CONCLUSIONKennedy disease usually occurs in mid-adulthood man. The clinical features are the weakness and wasting of limbs and bulbar muscles. Genetic analysis contributes to diagnosis and identification of carriers, and is beneficial to genetic counseling and prenatal diagnosis.

Adolescent ; Adult ; Aged ; Base Sequence ; Biopsy ; Bulbo-Spinal Atrophy, X-Linked ; diagnosis ; diagnostic imaging ; genetics ; pathology ; Child ; Child, Preschool ; Electromyography ; Exons ; genetics ; Female ; Genotype ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Muscles ; pathology ; Pedigree ; Receptors, Androgen ; genetics ; Ultrasonography ; Young Adult

Objective:To analyze the dosimetric differences between CT-guided free-hand intracavity/interstitial brachytherapy[image-guided adaptive brachytherapy (IGABT)] and conventional point-A plan (CP) in the treatment of cervical cancer.Methods:Twenty-six cervical cancer patients who received four cycles of IGABT in Sun Yat-sen University Cancer Center were enrolled in this study. Two sets of CT images were obtained before and after applicator adjustment to aid in the design of CP and IGABT plans. The high-risk clinical target volume (HRCTV), point A, and organs at risk (bladder, rectum, and sigmoid colon) were defined on CT images. CP and IGABT plans were designed on CT images. Parameter differences between CP and IGABT plans were analyzed with paired t-test and Wilcoxon test. Results:According to the coverage index (CI) of CP, plans were divided into two groups: in group A (CI≥0.90), 20 CP and corresponding IGABT plans were included, and 84 CP and corresponding IGABT plans in group B (CI<0.90). The mean volume of HRCTV and mean tumor diameter in group A were significantly smaller than those in group B (46.7 cm 3vs. 62.1 cm 3, P<0.001 and 3.1 cm vs. 4.4 cm, P<0.001). Compared with CP, IGABT significantly improved the value of D 90% in all plans and group B, whereas lowered the bladder dose. IGABT also reduced the dose of sigmoid colon in group A. IGABT significantly improved conformal index and dose homogeneity index. Conclusions:IGABT can significantly improve the target coverage, conformal index and dose homogeneity index, protect organs at risk. Compared with CP, IGABT has advantages in the treatment of patients with bulky tumor.

OBJECTIVETo investigate the expression feature of peroxiredoxin III in cervical lesions and to further understand the mechanism for cervical cancer development/progression.

METHODSExpression of peroxiredoxin III was immunohistochemically detected in cervical cancer. In addition, cervical epithelia were transfected with recombinant adeno-associated virus vector containing human papillomavirus 16 E6/E7 and peroxiredoxin III expression was detected by quantitative real time PCR and Western blotting.

RESULTSPeroxiredoxin III was significantly up-regulated in cervical cancer tissues. Nevertheless, expression of peroxiredoxin III remained unchanged in cervical epithelial cells after transfection.

CONCLUSIONIt seems that Prx III is not related to cervical cancer initiation. Up-regulation of peroxiredoxin III in cervical cancer might be an active response to oxidative stress in malignant cells, which protects against oxidatiton-induced apoptosis.

Cervix Uteri ; metabolism ; Female ; Gene Expression Regulation, Neoplastic ; Human papillomavirus 16 ; genetics ; metabolism ; Humans ; Middle Aged ; Oncogene Proteins, Viral ; genetics ; metabolism ; Papillomavirus E7 Proteins ; genetics ; metabolism ; Peroxiredoxins ; genetics ; metabolism ; Repressor Proteins ; genetics ; metabolism ; Up-Regulation ; Uterine Cervical Neoplasms ; genetics ; metabolism ; virology

OBJECTIVETo assess the antihypertensive effect and safety on medicine named 'Beijing Hypertensive No. 0' in a three-year treatment of primary hypertension.

METHODSA community-based intervention study was conducted. The antihypertensive effects and adverse events were observed.

RESULTS4000 patients with primary hypertension were randomly divided into two groups with 1529 patients treated with 'Beijing Hypertensive No. 0' and 976 patients treated with other antihypertensive drugs, among which 946 and 853 patients in the two groups completed the three-year study. After treatment, the systolic blood pressure decreased 13 mm Hg and 7 mm Hg while diastolic blood pressure decreased 8 mm Hg and 4 mm Hg in the 'No. 0' group and controlled group respectively. After three years of treatment, 90.0% and 79.5% in the 'No.0' group and in the control group had reached the BP 'fulfillment criteria', which were much higher than the baseline data. Side effects occurred in 33/1274 (2.6%) cases during three years' treatment with most commonly seen as dizziness, headache, palpitation and weakness. No serious adverse reactions occurred. There were some positive effects after treated by 'No. 0', including 0.13 mmol/L decrease of TC, 0.70 mmol/L decrease of LDL-C and an average 0.12 mmol/L increase of HDL-C. All of these changes were statistically significant. There were also opposite effects as 0.13 mmol/L increase of TG, 0.24 mmol/L increase of K+, and 0.88 mmol/L increase of Na+ on average, which were also statistically significant.

CONCLUSIONCompared with the conventional treatment, this treatment of 'Beijing Hypertensive No.0' was more convenient, safe and effective in treating mild to moderate primary hypertension in the community.

Aged ; Antihypertensive Agents ; adverse effects ; therapeutic use ; Female ; Follow-Up Studies ; Humans ; Hypertension ; drug therapy ; Male ; Safety