Objective: To study the application of glucose infusion test (GIT) in assaying the vascular access recirculation rate in hemodialysis patients. Methods: Access recirculation was assayed by both urea test (UT) and GIT in 82 hemodialysis patients, and 17 patients were also examined by Doppler ultrosonics method. The results of the Doppler ultrosonics were compared with those of UT and GIT. Results: Thirty-seven (45.12%) patients showed positive results with GIT and 29 patients (35.36%) with UT.All 17 patients had recirculation confirmed by Doppler ultrosonics and all had positive results by GIT (100%), but only 9 (52.94%) of the 17 patients had positive results by UT. Conclusion: Comapared with UT, GIT is more sensitive, more ecnomical,and simpler in determining vascular access recirculation, and can be used as a new method for screening vascular access recirculation.

To investigate the effects of low dosage of nitric oxide synthase (NOS) inhibitor Nc-nitro -L-arginine methyl ester ( L-NAME) in two-week treatment on the hyperdynamic circulatory state in rats with cirrhosis. METHODS: Cirrhosis model was induced in male SD rats by injection of 60 % CCL4 oily solution subcutaneously. Cirrhotic rats were treated with L-NAME ( 0.5 mg·kg-1·d-1) by gavage for two weeks. Mean arterial pressure ( AP ), portal pressure(PP), cardiac output ( CO ), cardiac index ( CI ), splanchnic vascular resistance ( SVR ), splanchnic blood flow(SBF) and serum nitrite levels were determined in L-NAME-treated, L-NAME-untreated cirrhotic rats and controls by using 57Co-labled microsphere technique and a fluorometric assay, respectively. RESULTS: Untreated cirrhotic rats had significantly lower MAP, SVR and higher PP, CO, CI, SBF and nitrite concentration than those of the controls (all,P< 0.01 ). In treated cirrhotic rats, L-NAME significantly attenuated the increase of CO, CI, SBF, nitrite concentration and the decrease of MAP and SVR. In treated cirrhotic rats, L-NAME induced a marked decrease of nitrite concentration than untreated cirrhotic rats[(1.471±0.907)μmol/L vs (4.204±1.253) μmol/L, P<0.01]. CONCLUSION: The endogenous NO may play an important role in the changes of hemodynamics pattern in cirrhosis, and hyperdynamic circulatory state in rats with cirrhosis can be ameliorated by oral two-week administration of lower dose of L-NAME.

Objective The purpose of this study was to evaluate the outcome of autogenous bone grafts in unilateral cleft lip and palate patients following early orthodontic tooth movement,and to determine the volume of new bone formation in the bone grafted region with spiral computed tomography.Methods Computed tomography scans of 12 patients were taken immediately preoperatively and at 6 months postoperatively.The patients underwent bone grafting between 9 and 13 years of age were divided into two groups based on whether postoperative orthodontic tooth movement were initiated or not.Three-dimensional models were created in each period,and the defect of alveolar cleft and volume of the newly formed bone were calculated in each patient.The roots of the moved teeth and their positions to the alveolar bone were also observed.Results The preoperative cleft width and cleft volume were not significantly different between both groups.The volume of the newly formed bone in group A was (0.98±0.23) mm3,significantly higher than that in group B,which was (0.73± 0.15) mm3.The rate of newly formed bone in group A was (72.5 ± 11.9)％,significantly higher than that in group B,which was (53.2±9.7)％.The cleft adjacent teeth could move smoothly into the bone grated area,with no root resorption observed in the computed tomography scans.Conclusions Early orthodontic tooth movement can reduce bone resorption in autogenous bone grafted unilateral cleft lip and palate patients through the observation of spiral computed tomography.It plays an active role in the bone remolding process after bone grafting.

With the development of sequencing technology, the cost of whole-genome sequencing was significantly declined.Meanwhile, with the application of combined whole-genome sequencing with epigenetic analysis on methylation and histone acetylation, the comprehensive and systematic analysis of numerous samples became a reality and we are able to re-understand the genesis and development of cancer. New ideas are emerging in comparative genomics research methods, from comparison of genomes among different individuals to horizontal self-comparison of different tissues and vertical self-comparison of genomes recently.Individualized diagnosis and treatment of cancer has shown a bright future.
Genome, Human ; genetics ; Humans ; Neoplasms ; genetics ; therapy ; Precision Medicine ; Sequence Analysis, DNA ; methods

OBJECTIVETo develop a solid phase PCR method by covalent single point immobilization for recycle utilization of human genome.

METHODSPolymethacrylamide gel was selected as a solid PCR carrier based on DNA-hydrogel copolymer chemistry presented by Mirzabekov. (CH2)6NH2 amino-modified PCR product and randomly fractured formic acid-modified plasmid pGEM-T-HLA-G were used as templates. The specificity of the attachment chemistry was characterized by acrylamide gel electrophoresis, and the thermal stability of method was demonstrated by PCR. This method was applied for the recycle utilization of human genome. Sequencing was used to exclude the possibility of introduced mutations during modification and immobilization procedures.

RESULTSThe PCR detections of plasmid DNA and human genome DNA immobilized by polymethacrylamide gel was successful. The thermal stability of method was successfully demonstrated by performing PCR after 16 rounds of standard 36 PCR cycles. And the sequencing was found no mutation.

CONCLUSIONThe DNA immobilization method with polymethacrylamide gel as a solid phase carrier is stable and specific, which can be a possible approach for realizing recycle utilization of human genome for whole-genome sequencing and SNP detection.

Electrophoresis, Polyacrylamide Gel ; Genome, Human ; Humans ; Hydrogels ; Immobilized Nucleic Acids ; analysis ; Polymerase Chain Reaction ; methods

BACKGROUND: The neurotoxicity and cardiotoxicity of bupivacaine have been reported frequently. However, the studies on bupivacaine-induced muscle toxicity are few.
OBJECTIVE: To establish and evaluate local intramuscular injection of bupivacaine on the changes in histomorphology and ultrastructure of rat multifidus muscle at various time points.
METHODS: A total of 54 male Sprague-Dawley rats weighing 280-320 g were randomly divided into black group (n=18), model group (n=18) and model control group (n=18). Each group was then equal y subdivided into three subgroups according to time points (4, 7 and 14 days) (n=6). Both sides of multifidus muscle of the rats (L4 and L5) were injected with 0.5% bupivacaine. The morphological and ultrastructural changes of multifidus muscle were observed and analyzed with light microscope and transmission electron microscope at 4, 7 and 14 days after model establishment.
RESULTS AND CONCLUSION: (1) A single intramuscular injection of 0.5% bupivacaine induced muscular damage. (2) Hematoxylin-eosin staining results showed fiber necrosis, inflammatory cel infiltration, and a smal amount of macrophages in local skeletal muscle. (3) Under the transmission electron microscope, the structure of myofibrils was destroyed or disintegrated; kinds of bands and lines were indistinct, disrupted or disappeared; the structure of mitochondria was abnormal, the mitochondrial cristae were reduced or disappeared. In the 7- and 14-day groups, multifidus muscle proliferated and repaired. (4) Ultrastructural change scores in skeletal muscle were significantly higher in the model group than in the blank and model control groups (P < 0.05). Above scores were significantly greater in the 4-day group than in the 7- and 14-day groups (P < 0.05), and higher in the 7-day group than in the 14-day group (P < 0.05). (5) Results suggest that a single intramuscular injection of 0.5% bupivacaine can result in pathological changes of skeletal muscle from morphology and ultrastructure. This method can establish a suitable model of skeletal muscle injury.

Objective To investigate the value of nerve conduction studies (NCSs),F wave analysis,somatosensory evoked potential (SEP) and skin sympathetic response (SSR) in the early diagnosis of diabetic peripheral neuropathy (DPN).Methods A total of 110 patients with diabetes mellitus were recruited as the diabetic group and another 50 well-matched healthy volunteers as the normal controls.Sensory and motor NCSs of the median,ulnar,posterior tibial and common peroneal nerves were performed.F waves were recorded from the median and posterior tibial nerves.SEPs elicited by stimulation to nerves of both the upper and lower limbs as well as SSRs were measured,all in both the diabetic group and the normal controls.Results The total rate of nerve conduction abnormality was 74.5％ in the diabetic group,with sensory nerve conduction abnormalities more frequent and more severe among motor nerves in the extremities.The total rate of F wave abnormalities was 57.3％ in the diabetic group.The rate in patients with normal distal motor conduction in their median and posterior tibial nerves was 50.7％.The total SEP abnormality rate was 70.0％ with regard to the proximal peripheral nerve potentials in the diabetic group,but there was no obvious abnormality of the supraclavicular electrical potential in the upper limbs for those with normal sensory nerve conduction in the median nerve.The rate of occurrence of abnormality in the gluteus point potential in the lower limbs of those with normal posterior tibial sensory conduction was 62.5％.The total rate of SSR abnormalities was 80.0％ in the diabetic group but 72％ among those with normal nerve conduction in their extremities.Combining the NCS,SSR,SEP and F wave results,the total abnormality rate was 90.9％ in the diabetic group,which was much higher than with any single test used alone.Conclusion NCS is essential for diagnosing DPN.Early diagnosis of subclinical diabetic neuropathy will be significantly enhanced when nerve conduction,SSRs,SEPs and F waves are tested together.

Recent years, the incidence and mortality of prostate cancer have increased dramatically in China. At earlier stages, most diagnosed prostate cancers are responsive to androgen depletion treatment, yet, nearly all patients will eventually progress to metastatic androgen-independent prostate cancer (AIPC), which still has no effective therapeutic method or drug to deal with. 11'-Deoxyverticillin A (C42) belongs to the family of epipolythiodioxopiperazines (ETPs), an interesting class of fungal toxins that inhibit farnesyl transferase. Compounds holding such a property have been explored as putative anticancer agents. In this study, using PC3M cells, an AIPC cell line, we investigated the effect of the compound on apoptosis and explored the underlying mechanism. It revealed that C42 markedly enhanced the activity of caspase-3/7 and increased the accumulation of the cleaved PARP, all of which are the markers of apoptosis. It also revealed that C42 either decreased cell viability or inhibited the growth of PC3M cells. Moreover, we observed that the loss of cell viability and cell growth inhibition induced by C42 were both time- and dosage dependent. Taken together, we indicated that C42 can induce caspase-dependent apoptosis in AIPC cells, and the results presented here will broaden our knowledge about the molecular mechanisms by which C42 exerts its anticancer activity, and future work in this direction may provide valuable information in the development of these compounds into effective cancer therapeutic strategies against androgen-independent prostate cancer.
Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Caspase 7 ; metabolism ; Cell Line, Tumor ; Disulfides ; pharmacology ; Farnesyltranstransferase ; antagonists & inhibitors ; Humans ; Male ; Mycotoxins ; pharmacology ; Piperazines ; pharmacology ; Prostatic Neoplasms ; pathology

OBJECTIVETo analyze the deletion region for two fetal cases with large Yq deletions in order to provide genetic counseling and prenatal diagnosis.

METHODSFor both cases, amniotic fluid samples were cultured and analyzed with G banding and fluorescence in situ hybridization (FISH). Multiplex polymerase chain reaction was also carried out to amplify 15 sequence tagged sites (STS) of azoospermia factor (AZF) on the Y chromosome.

RESULTSFor both samples, the karyotypes were determined as 46,X,del(Y)(pter→q11:). No heterochromatin was found in C band. The karyotypes of their fathers were 46,XY, and heterochromatin was found in C band. STS analyses suggested that only sY82, sY84 and sY86 in AZFa were amplifiable while the other 12 STS were negative in amniotic fluid for the first case, which indicated deletions of AZFb, AZFd and AZFc. No AZF deletion was found in its father. For the second case, all 15 STS were amplifiable in the amniotic fluid, suggesting no AZF deletion. No AZF deletion was found in its father too.

CONCLUSIONConventional karyotyping combined with FISH and molecular genetics techniques can enable characterization of AZF microdeletions and facilitate genetic counseling and prenatal diagnosis.

Adult ; Azoospermia ; genetics ; Chromosome Deletion ; Chromosomes, Human, Y ; genetics ; Female ; Fetal Diseases ; diagnosis ; genetics ; Genetic Counseling ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Male ; Pregnancy ; Prenatal Diagnosis

OBJECTIVETo track and analyze two false positive cases from non-invasive prenatal testing for potential fetal aneuploidy.

METHODSThe two cases, respectively reported to have XO (+++) and T18 (1/20) XO(+), were analyzed with conventional karyotyping, fluorescence in situ hybridization (FISH) and massively parallel genomic sequencing (MPS).

RESULTSThe first fetus, who was suspected for XO(+++), was verified to have super female syndrome (47,XXX/46,XX) due to confined placental mosaicism by karyotyping of amniotic fluid cells, FISH analysis of placenta and massively parallel sequencing (MPS) of fetal tissue. The second fetus, suspected to have trisomy 18 (1/20) XO(+), was verified to have Turner syndrome by karyotyping, FISH and MPS analyses of umbilical cord blood cells. And the karyotype was 45,X[48]/46, X, der(X) del(X) (p11.21) del(X) (q13.3)[62].

CONCLUSIONNon-invasive prenatal testing carries a risk for false positive diagnosis of fetal sex chromosome and trisomy 18. Combined cytogenetic and molecular techniques are required to ensure an accurate diagnosis.

Adult ; Aneuploidy ; Chromosome Aberrations ; Diagnostic Errors ; False Positive Reactions ; Female ; Fetal Diseases ; diagnosis ; genetics ; Humans ; Pregnancy ; Prenatal Diagnosis ; Young Adult