How to get functional gene from uncultured-microbiology is the hotspot content of microbial ecology. What the most important is how to obtain the pure and integrated genomic DNA. An efficient, nonselective extraction method to gain chromosomal DNA from eight kinds of bacteria was introduced. Amount DNA released by hot-detergent gave the highest DNA yields from different G + and G- bacteria. Running 20 hours by PFGE mode, the size of total DNA is over 23kb. The pure DNA could be digested by Hind Ⅲ and used in PCR. The total environmental DNA also can be extracted from soil by the same method. As a result it showed a new way for the environmental DNA extraction.

Methods for studying the population diversity of microorganism in activated sludge usually require enrichment of bacterial genome.The efficient information on microbial species composition provided and shifted in diversity revealed are dependent on the effective DNA recovery technique.The method was based on washing by alkaline phosphate buffer and digestion with extended heating of the activated sludge suspension in the presence of lysozyme and freeze-thawing in high-salt-SDS buffer.The extraction was tested for four activated sludge differing in places and dates.The DNA fragment from all sludge was integrity.DNA yields ranged from 105 to 823 ?g/g sludge and were of sufficient purity for PCR-based 16S ribosomal DNA analysis and restriction digested.In general,all methods produced DNA pure were not enough for PCR amplification and libraries construction.As basis of experimental goals,the study provides an appropriate extraction method of microbial DNA in sludge.

The composition of intestinal microflora is closely related to the occurrence and development of colorectal cancer (CRC). Among them, Fusobacterium nucleatum (Fn) has been proved directly related to the recurrence, metastasis and chemotherapy resistance of CRC. Therefore, it is of great significance for the prevention and treatment of colorectal cancer by the exploration potential anti-Fn drug targets and discovery small molecule drugs. However, no selective anti-Fn small molecule inhibitors have been reported so far as well as their anti-Fn thereby "anti-Fn further anticancer" mechanisms are unclear. Herein, this article reviews the potential therapeutic targets and small molecule ligands of Fn in order to provide a reference for the development of anti-Fn and anti-CRC small molecule drugs.

The overexpression of doublecortin-like kinase 1 (DCLK1) is closely related to the occurrence and development of various malignant tumors. Discovery of novel anti-tumor agents targeting DCLK1 represents a hot spot in this field. So far, several DCLK1 small molecule inhibitors with excellent anti-tumor activity in vitro and in vivo have been designed and synthesized, which is expected to provide a new strategy for tumor therapy. This article reviews the research progress in the discovery, structure type, structural optimization, biological activity and mechanism of action of DCLK1 small molecule inhibitors, and provides research basis for the development of new anti-tumor small molecule inhibitors targeting DCLK1.

Natural products are important sources for the discovery of anti-tumor drugs. Evodiamine is the main alkaloid component of the traditional Chinese herb Wu-Chu-Yu, and it has weak antitumor activity. In recent years, a number of highly active antitumor candidates have been discovered with a significant progress. This article reviews the research progress of evodiamine-based antitumor drug design strategies, in order to provide reference for the development of new drugs with natural products as leads.

BACKGROUND: Although the pathogenesis of vitiligo isn't fully understood, a recent study demonstrates that oxidative stress plays an important role to induce vitiligo. Peroxiredoxin (Prx) is a novel peroxidase family to remove hydrogen peroxide using thioredoxin system, which is consisted of thioredoxin, thioredoxin reductase, and NADPH. OBJECTIVE: This study aimed to investigate the change of expression of Prx I to elucidate the role of oxidative stress in the pathogenesis of vitiligo. METHODS: Sample specimens were obtained from the lesional skin of vitiligo patients, and non-depigmented skin was obtained from the perilesional area as control samples. The skin samples were immediately frozen using liquid nitrogen, and then section samples were prepared to perform immunohistochemical staining with antibodies for Prx I. Some of the skin biopsy samples were used for primary culture of keratinocytes. Protein extracts from the expanded keratinocytes were prepared for Western blot analysis of Prx I. RESULTS: In vitiligo, the ubiquitous expression of Prx I in all layers of epidermis, which was also observed in the normal perilesional skin, was reduced in the depigmented lesion of vitiligo patients. The reduction of Prx I was remarkable from the lesions which were exposed to sunlight. Consistently, Prx I expression from the lesional keratinocytes were noticeably reduced in comparison with that from perilesional keratinocytes. CONCLUSION: Our results showing that Prx I is impaired in the epidermis of depigmented lesions of vitiligo patients suggest that oxidative stress is an important factor to induce vitiligo.
Antibodies ; Biopsy ; Blotting, Western ; Epidermis ; Humans ; Hydrogen Peroxide ; Keratinocytes ; Nitrogen ; Oxidative Stress ; Peroxidase ; Peroxiredoxins ; Skin ; Sunlight ; Thioredoxin-Disulfide Reductase ; Thioredoxins ; Vitiligo

Adult ; Humans ; Methylprednisolone ; therapeutic use ; Optic Neuritis ; diagnosis ; drug therapy ; etiology ; Spondylitis, Ankylosing ; complications

Objective To describe the epidemiological and serological features on a family associated outbreak caused by Mycoplasma pneumoniae (MP) infection occurred in Beijing in August 2007.Methods Mutual exposure of the family members was investigated and retrospective medical record was reviewed for the hospitalized patients.Serum antibodies to MP were measured and chest X-rays were taken for all the family members.Results This family consisted of 5 members,with fixed members as the boy (13 years old ),his father (43 years old) and mother (44 years old),grandmother (64 years old) and uncle (32 years old ) who was involved in taking care of the sick boy and his father.During 23 days of the event,four of all the five family members were ill.Three (boy,father and uncle) had radiographic pneumonia,whose paired sera all showed a ≥ fourfold increase in antibody titer,and two of them were confnrmed by chest X-ray on day 2 after onset of fever.The grandmother suffered from bronchitis,with positive(PA) serum antibody to MR Serum MP-IgG from the father and uncle was positive,3 days and 2 days after the onset of fever.The chances of contact between grandmother with the boy and uncle with the father were both only in the hospital wards.Only the mother remained asymptomatic,with her serum MP-IgM (-)and MP-IgG ( + )for which the blood sample was collected 37 days after close contact with the boy.The longest time of exposure to the patients was between mother and the boy but only the mother did not increase her total workload or feeling for fatigue.Conclusion Results of MP-IgG from post-infection did not completely defend against the repeated MP infection.Combined risk factors as index patients with severe cough,prolonged close contact,poorly ventilation of the environment,and family members with excessive fatigue might work as the causes of this family MP outbreak.

Sleep is an important component in our lives as it is necessary throughout one’s entire life span. This study was conducted to elucidate whether there are gender differences in sleep quality and what factors can affect sleep quality in community-dwelling elderly Koreans. A total of 382 subjects (175 males and 207 females) were recruited among elderly aged 45 or over who participated in the 2010 Hallym Aging Study (HAS). They were invited to a general hospital and were evaluated for socioeconomic status, smoking history, and various clinical measures. Sleep quality was assessed by the Pittsburgh Sleep Quality Index (PSQI). A higher score indicates poorer subjective sleep quality, (PSQI global score > 5 suggests sleep disturbance). After adjusting for potential covariates, our results show that alcohol increases the odds for poor sleep (odds ratio [OR] = 3.35, 95% confidence interval [CI] = 1.11–10.10) in males. In females, lack of exercise was the major risk factor of poor sleep as they are 4.46 times more likely to suffer from low sleep quality than those who exercise regularly (95% CI=1.56–13.75). Stress was also a risk factor for poor sleep. It was 5.60 times higher in the "always have stress" group than the "do not have stress" group (95% CI = 1.54–20.34). Thus, alcohol consumption is associated with men’s sleep quality, while exercise and stress level affect women's.
Aged* ; Aging* ; Alcohol Drinking ; Female ; Hospitals, General ; Humans ; Male ; Risk Factors ; Smoke ; Smoking ; Social Class

OBJECTIVETo investigate the clinical therapeutic results of allograft tendon for anatomical reconstruction of medial patellofemoral ligament (MPFL) for the treatment of patellar dislocations.

METHODSFrom September 2008 to June 2013, 16 patients with patellar dislocation underwent MPFL reconstructions. There were 2 males and 14 females, aged 11 to 27 years old (16 years old on average). Patellar dislocations occurred in 11 left and 5 right knees. The disease course ranged from 3 to 10 years. The frequency of dislocation ranged from 9 to 33 times (19 times on average). Affected knee joints showed patellar instability; the range of action for patella obviously increased. The X-ray films showed patellar dislocation. The preoperative Q angle was (36 ± 9)°, and the congruence angle was (63 ± 18)°. Reconstruction was performed via allograft tendon. Allograft tendon was fixed through the superomedial pole of the patella, and the other end was fixed at the natural MPFL insertion site near the medial femoral condyle with an interference screw in a bone tunnel. All the patients were evaluated postoperatively; Kujala patellofemoral scores, objective knee function, complications, and reoperations were assessed.

RESULTSPrimary healing was achieved in all cases. No infection or necrosis and absorption of grafts was observed. All the patients were followed up for an average of 16.4 months (ranged, 10 to 24 months) postoperatively. At the latest follow-up, all the patients had no pain, swelling and patellar instability; neither patella redislocation nor fracture occurred. The X-ray films showed good position of tunnel 6 months after operation, and the congruence angle was (5 ± 9)°, showing statistically significant difference when compared with preoperation (P < 0.05). The postoperative Q angle was (17 ± 8)°, the Kujala knee function score improved significantly from 45.20 ± 9.20 to 89.30 ± 6.40 at the latest follow-up, showing statistically significant difference (P < 0.05).

CONCLUSIONMPFL reconstruction improves clinical symptoms. Anatomical MPFL reconstruction is effective for patellar dislocation, and it offers good recovery of the premorbid patella mechanics. The interference screw provides firm fixation. Allograft can avoid the graft harvest site morbidity, but it increases the cost of the surgery.

Adolescent ; Adult ; Allografts ; Child ; Female ; Humans ; Ligaments, Articular ; surgery ; Male ; Patellar Dislocation ; surgery ; Patellofemoral Joint ; surgery ; Reconstructive Surgical Procedures ; methods ; Tendons ; transplantation