Adult ; Bloodletting ; Combined Modality Therapy ; Female ; Herpes Zoster ; therapy ; Humans ; Male ; Manipulation, Chiropractic ; Middle Aged ; Young Adult

Acupuncture Therapy ; Acute Disease ; Adolescent ; Adult ; Child ; Combined Modality Therapy ; Conjunctivitis ; therapy ; Female ; Humans ; Male ; Middle Aged ; Needles ; Phlebotomy ; instrumentation

Acupuncture Therapy ; Adult ; Aged ; Aged, 80 and over ; Cold Temperature ; Female ; Humans ; Male ; Meridians ; Middle Aged ; Qi ; Yin Deficiency ; therapy

Objective To investigate the clinical,neuroimaging and myopathological features of mitochondrial encephalomyopathy with lactic acidosis and stroke-like episodes(MELAS).Methods The clinical manifestations,neuroimaging and myopathological features of 31 patients with MELAS diagnosed in our Neuromuscular Center in the recent 7 years were retrospectively analyzed.A3243G point mutations were analyzed by RFLP method in 10 patients.Results ①Clinical features:There were 18 male patients and 13 female patients.The age of onset ranged from 3 to 43 years,averaging 21.9 years.The averaged duration was 4.9 years.Thirteen patients in this group had family history of maternal inheritance pattern.The main clinical manifestations included short stature(26 patients),recurrent headache and vomiting(24 patients), muscle weakness(22 patients),epileptic seizure(21 patients),cognitive decline(19 patients),visual disturbance(17 patients),sensorineural deafness(16 patients),ataxia(6 patients),psychiatric symptom (8 patients),external ophathalmoplegia(2 patients)and diabetes mellitus(9 patients).The serum CK level was slightly elevated in 6 patients,and the fasting blood lactic acid was increased in 15 of the 18 detected patients.②Neuroimaging features:The stroke-like lesions were mostly confined to cerebral cortex, including temporal lobe(24 patients),occipital lobe(21 patients),parietal lobe(12 patients)and frontal lobe(4 patients).Three patients had deep white matter involvement.Migrating stroke-like lesions were confirmed in 4 patients by repeated cranial CT/MRI examination.In addition,cerebral atrophy(17 patients)and bilateral basilar ganglion calcification(11 patients)were found.③Myopathological features: Scattered ragged red fibers(RRF)in various number were found in all the patients by MGT staining.Other founding included strongly SDH-reactive blood vessel(27 patients),COX enzyme deficiency(19 patients), and mild to moderate lipid storage in RRF(20 patients).④MtDNA analysis showed 9 patients with A3243G point mutation in all the detected 13 patients.Conclusion The clinical and neuroimaging features may offer important clue to the diagnosis of MELAS,but a definite diagnosis of MELAS relies on the myopathology and mtDNA mutation analysis.

Fruit ripening is associated with a number of physiological and biochemical changes. They include degradation of chlorophyll, synthesis of flavor compounds, carotenoid biosynthesis, conversion of starch to sugars, cell wall solublisation and fruit softening. These changes are brought about by the expression of specific genes. People are interested in the molecular mechanism involved in the regulation of gene transcription during fruit ripening. Many fruit-specific promoters such as PG, E4, E8, and 2A11 have been characterized and shown to direct ripening-specific expression of reporter genes. AGPase plays the key role in catalyzing the biosynthesis of starch in plants. It is a heterotetrameric enzyme with two small subunits and two large subunits, which are encoded by different genes. In higher plants, small subunits are highly conserved among plant species and expressed in all tissues. And the large subunits are present at multiple isoforms and expressed in a tissue-specific pattern. In fruits, the expression pattern of the large subunits varies with plant species. That made it important to study the transcriptional regulation of the large subunits of AGPase in different plant species. Northern-blot analysis indicates in watermelon, an isoform of the large subunits Wml1 expressed specifically in fruits, not in leaves. The 5' flanking region of Wml1, which covers 1573bp, has been isolated through the method of uneven PCR. And transient expression assay has shown that the 1573bp (named WSP) can direct fruit-specific expression of GUS gene. Our goal in this study was to scan the promoter region for main regulatory regions involved in fruit-specific expression. A chimaeric gene was constructed containing the WSP promoter, the beta-glucuronidase (GUS) structural sequence as a reporter gene and the nopaline synthase polyadenylation site (NOS-ter). The plasmid pSPA was digested with Hind III + Hinc II and promoter fragment of 1573bp (from 180bp to 1752bp) was cut out and cloned into Sma I sites of pBluescript SK(-), to produce pBSPA-16. The same insert was then cut out with Hind III + BamH I, and ligated with transient expression vector pBI426 digested by HindIII + Bgl II to produce pISPA-16. Three 5'-end deletions of the promoter were obtained and fused to GUS gene in plant transient expression vector pBI426: the 1201bp fragment (from 551bp to 1752bp) was generated by digestion of pBSPA-16 with BamH I + SnaB I, the 898bp fragment (from 854bp to 1752bp) by BamH I + EcoRV. Both fragments were ligated with pBluescript SK(-) digested by BamH I + Sma I, to produce pBSPA-12 and pBS-PA-9. The inserts were cut out with HindmIII + BamH I and ligated with pBI426 digested by Hind III + Bgl II, to produce pISPA-12 and pISPA-9. The 795bp fragment (from 957bp to 1752bp) was generated by digestion of pSPA with Hinc II + EcoR I, promoter fragment was cut out and cloned into Sma I sites of pBluescript SK(-), to produce pBSPA-8. The same insert were cut out with Hind III + BamH I, and ligated with transient expression vector pBI426 digested by Hind III + Bgl II. The 1573bp fragment and three 5'-end deletions were delivered into watermelon leaf, stem, flower and fruit of different development stages (5, 10, 20 days after pollination) via particle bombardment using a biolistic PDS-1000/He particle gun. Bombardment parameters were as follows: a helium pressure of 1200 psi, vacuum of 91432.23Pa, 7 cm between the stopping screen and the plate. Histochemical assay were done on all the tissues bombarded after incubation for 2 days. The 1573bp fragment had the strongest promoter activity, and can induce GUS expression in fruits of 5 and 20 days after anthesis and flowers, but not in fruits of 10 days after anthesis, leaves and stems. Fragments of 1201bp and 898bp can induce GUS expression only in fruits of 20 days after anthesis, and with lower expression levels than 1573bp. Fragment of 795bp was not able to direct GUS expression in any of the tissues bombarded (data not shown). It can be concluded that of the 1573bp, 1201 bp, 898bp Wml1 5'flanking regions include the necessary information directing fruit-specific expression. Deletion from 180bp to 551bp doesn't affect the fruit-specificity of the promoter, but lowered the expression level. There may be some cis-acting elements located in this region, which can enhance external gene expression in later stages of fruit development. Deletion from 854bp and 958bp led to loss of GUS expression. This region includes the necessary information needed for gene expression as well as the regulatory elements for fruit-specific transcription.
Citrullus ; genetics ; Fruit ; genetics ; Gene Expression Regulation, Plant ; genetics ; Promoter Regions, Genetic ; genetics ; Regulatory Sequences, Nucleic Acid ; genetics ; physiology

OBJECTIVETo investigate the incidence of lymph node metastasis (LNM) in early esophageal carcinoma and the techniques of dissection.

METHODSStandard three-field dissection was performed in patients with small superficial esophageal carcinoma detected by endoscopy from 1993 - 2007. The lymph node metastases in different regions were identified by histopathology. The survival rate of the cases was analyzed.

RESULTSA total of 149 patients with early esophageal carcinoma were identified by postoperative pathological examination. The overall lymph node metastasis (LNM) rate was 22.8%, and the degree of LNM was 2.4% in all fields. Most lymph node metastases from upper thoracic esophageal carcinoma were found in cervical and the right upper mediastinal nodes. The LNM from middle thoracic esophageal carcinoma were approximately equal in the cervical, mediastinal, and abdominal lymph nodes, and abdominal lymph node metastasis predominated in lower thoracic esophageal carcinoma. The metastatic rate of LNM adjacent to the right recurrent laryngeaal nerve was the highest (44.1%). Significant differences were shown among the rates of LNM in relation to different macroscopic pattern, depth of invasion and differentiation of tumor (P < 0.01), but not to the longitudinal length of tumor (P > 0.05). The overall 5-year survival rate was 77.9%. It was 87.0% in patients without LNM, and 47.1% in those with LNM.

CONCLUSIONLymph node metastasis in early esophageal carcinoma is in a high frequency. Patients with tumor invasion into the mucosa or lamina propria but without lymph node metastasis may undergo a local operation such as endoscopic mucosectomy and have a good prognosis. Patients with tumor invasion into the muscularis mucosae or submucosa should be treated with radical surgery with three-field lymphadenectomy, especially, to dissect the lymph nodes adjacent to the recurrent laryngeal nerve.

Abdomen ; Adult ; Aged ; Carcinoma, Squamous Cell ; pathology ; surgery ; Esophageal Neoplasms ; pathology ; surgery ; Female ; Follow-Up Studies ; Humans ; Lymph Node Excision ; methods ; Lymph Nodes ; pathology ; surgery ; Lymphatic Metastasis ; Male ; Mediastinum ; Middle Aged ; Mucous Membrane ; pathology ; Neck ; Neoplasm Invasiveness ; Neoplasm Staging ; Survival Rate

OBJECTIVETo compare the prognosis between staged surgery and one-stage surgery in active complex Crohn disease(CD).

METHODSClinical data of 33 patients with active complex CD from February 2006 to September 2010 were analyzed retrospectively.

INCLUSION CRITERIAPatients who were diagnosed with CD by pathology or endoscopy with CD activity index(CDAI)>220 and long history of preoperative steroid use(over 6 months), or complicated with severe preoperative malnutrition. The indications for surgery included enterocutaneous or entero enteric fistula, with/without intra-abdominal abscess, intestinal obstruction, and acute intestinal perforation intra-abdominal sepsis. The surgical procedures, including staged surgery and one-stage definite surgery were chosen by experienced surgeons. All the patients quit smoking and received immunosuppressant therapy (TwHF or azathioprine) together with enteral nutrition supplement for preventing postoperative recurrence. All the patients were followed up after the surgery with regards to CDAI, serum C-reactive protein (CRP), erythrocyte sedimentation rate (ESR). Postoperative complications, clinical and endoscopic recurrence were recorded.

RESULTSOf the 33 patients, 14 underwent one-stage definite surgery and 19 received staged surgery. Postoperative complication rates in the two groups were 71% and 26% respectively(P=0.015). The early clinical recurrence rates within postoperative 3 months were 36% and 0(P=0.008). In regard to cumulative probability of post-operative clinical and endoscopic recurrence, staged surgery group was superior to one stage definite surgery group(P=0.000 and 0.006).

CONCLUSIONStaged surgery may significantly increase the success rate of operation, reduce postoperative complications and postoperative early relapse, and decrease postoperative recurrence.

Adolescent ; Adult ; Aged ; Crohn Disease ; surgery ; Digestive System Surgical Procedures ; methods ; Female ; Follow-Up Studies ; Humans ; Logistic Models ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome ; Young Adult

OBJECTIVETo investigate the efficacy of enteral nutrition(EN) therapy for active Crohn disease(CD) complicated with incomplete intestinal obstruction.

METHODSClinical data of 37 patients with active CD complicated with incomplete intestinal obstruction treated with EN(n=37) between January 2003 and September 2009 were retrospectively analyzed. CD activity index (CDAI) was between 150 and 450. The patients received total enteral nutrition (TEN) 125 kJ/kg by nasogastric tube or percutaneous endoscopic gastrostomy/jejunostomy(PEG/J) tube. Clinical response was defined as a decrease in CDAI≥70 from baseline since EN therapy, and clinical remission was defined as CDAI<150. Nutritional status, disease activity index, and side effects were recorded at the 0, 4th, and 12th week after EN therapy.

RESULTSStricture or stenosis location included ileum in 8 (21.6%) patients, ileocolon in 19(51.4%), colon in 4(10.8%), jejunoileum in 5(13.5%), and duodenum in 1(2.7%). At 4 weeks after EN, CDAI significantly decreased(112.0±39.6 vs.174.6±34.7,P<0.05). The ratio of clinical response was 43.2%(16/37) and clinical remission was 72.9%(27/37). At 12 weeks, CDAI was 70.2±32.9, lower than that at week 4(P<0.05). The ratio of clinical response was 70.2%(26/37) and clinical remission was 78.4%(29/37). Other disease activity indexes such as C-reactive protein, erythrocyte sedimentation rate, and nutritional status such as BMI, serum albumin, prealbumin, transferrin and hemoglobin showed similar trend. During therapy, 7 cases had progressive intestinal obstruction resulting in bowel resection, 11 cases had diarrhea and/or abdominal distention due to inadequate infusion of home EN whose symptoms were improved after correction by the doctor.

CONCLUSIONSEN therapy can induce clinical response and remission in CD complicated with incomplete intestinal obstruction, relieve obstruction, alleviate the inflammatory response which plays positive role in the treatment of CD.

Adult ; Aged ; Crohn Disease ; complications ; therapy ; Enteral Nutrition ; Female ; Humans ; Intestinal Obstruction ; complications ; therapy ; Male ; Matched-Pair Analysis ; Middle Aged ; Retrospective Studies ; Treatment Outcome ; Young Adult

OBJECTIVETo study the effect of gene expression of mouse uroplakin II (UPII) promoter on human bladder cell cancer cell line.

METHODSThe mRNA expression of different cell lines was quantified by RT-PCR. Green fluorescent protein (GFP) and luciferase (Luc) were used as reporter genes. The plasmids carrying UPII or GFP were constructed and transfected into human cell lines of bladder transitional cell cancer (BIU-87), kindey cancer (GRC-1), vascular endothelium (EC), lung cancer cell line (A549) and skin fibroblast cell line (Hs27). GFP activity of cells was detected by confocual microscopy and flow cytometry (FCM). Luciferase value was measured by luminometer (microplate) and luciferase to beta-galactosidase ratios (L/G values) were used for evaluating transfection efficiency.

RESULTSRT-PCR showed high expression level of UPII mRNA in bladder cancer cell line BIU-87, whereas low level or no expression in nonbladder cancer cell lines. The activity of GFP in bladder cancer (BIU-87) cell was higher than that in the other cell lines (5 - 10/HP versus 0 - 2/HP), with 4.34% positive cells in BIU-87 detected by FCM, but no positive cell was found in the other cell lines. L/G values indicated that the luciferase expression in human bladder cancer cells transfected with mouse UPII promoter was 1.8 - 8.2-fold as high as that in the nonbladder cell lines.

CONCLUSIONMouse UPII promoter gene can be expressed in a tissue-specific fashion in human urinary bladder cancer. It is capable of initiating transcription of reporter genes in human bladder cancer cell line.

Animals ; Cell Line, Tumor ; Flow Cytometry ; Genetic Therapy ; Green Fluorescent Proteins ; Humans ; Luminescent Proteins ; genetics ; Membrane Proteins ; genetics ; Mice ; Organ Specificity ; Promoter Regions, Genetic ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection ; Urinary Bladder Neoplasms ; genetics ; therapy ; Uroplakin II

OBJECTIVETo investigate the effect of anastomosis suture on the postoperative recurrence in patients with Crohn disease(CD).

METHODSA total of 102 patients undergoing intestinal resection for CD at the Jinling Hospital of Nanjing University School of Medicine from Jan. 2002 to Jan. 2010 were studied retrospectively. The postoperative recurrence(endoscopic and clinical) in patients receiving intestinal anastomosis with VICRYL absorbable suture(n=48) was compared with that of patients sewn with normal silk suture(n=54).

RESULTSThe 1-, 2-, and 3-year accumulated clinical recurrence rates in the absorbable suture group and silk suture group were 6.3%, 14.6%, 22.9%, and 18.5%, 27.8%, 35.2%, respectively. The differences were not statistically significant(P=0.213). The 1- and 2-year accumulated endoscopic recurrence rates in the absorbable suture group and silk suture group were 14.7%, 38.2% and 22.9%, 62.9%, respectively. The differences were statistically significant(P=0.034).

CONCLUSIONSCompared to silk suture, use of absorbable suture for anastomosis may lower the risk of postoperative endoscopic recurrence after bowel resection in patients with CD. Absorbable suture should be the first choice for intestinal anastomosis in CD patients.

Adult ; Anastomosis, Surgical ; methods ; Crohn Disease ; etiology ; surgery ; Female ; Humans ; Male ; Middle Aged ; Recurrence ; Retrospective Studies ; Sutures ; Treatment Outcome