[A case of diabetic foot treated with fibula transversal bone transfer and micro-vascular network regeneration].

Reports a case of diabetic foot patient with right toe pain, ulceration for 4 years and aggravation for 1 month. After rigorous medical treatment and related examinations, transeversal bone transfer of tibia was proposed. However, the skin condition on the inner side of the lower leg was not good before the operation, so the transversal bone removal of the fibula was performed. Two segments of the lateral fibula were osteotomized with miniature osteotomies. The two osteotomy lines were about 10 cm apart. The distal osteotomy line is about 10 cm from the tip of the lateral malleolus. On the 8th day after the surgery, the bone was moved every 6 hours and was extended inwards by 1 mm every day. After 14 days, the bone was moved back for another 14 days. Skin temperature from preoperative to postoperative, ankle brachial index, CT angiography, CT value, visual analogue scale (VAS) of pain and wound healing were evaluated by comparison. The wound was completely healed 28 days after the operation.

Fibroblasts isolated from a keratocystic odontogenic tumor promote osteoclastogenesis in vitro via interaction with epithelial cells.

OBJECTIVES: Investigate the role of the epithelial-mesenchymal interaction of keratocystic odontogenic tumor (KCOT) in influencing osteoclastogenesis. MATERIALS AND METHODS: Fibroblasts isolated from KCOT fibrous capsule and normal gingival mucosa were, respectively, co-cultured with human immortalized oral epithelial cells (HIOECs), and the supernatant was collected to make conditioned medium, in which the osteoclastogenesis of osteoclast precursor cell line Raw 264.7 was observed. Genes related to bone resorption (RANKL, OPG, COX-2, and M-CSF) were analyzed by real-time PCR. Antibodies against human sRANKL and inhibitor of COX-2: NS398 were added to conditioned medium to investigate the inhibitory effect on osteoclastogenesis. RESULTS: Compared with co-cultured gingival fibroblasts and HIOECs (GE-CM), the conditioned medium from co-cultured KCOT fibroblasts and HIOECs (KE-CM) induced more osteoclast-like cell formation and increased NFATC1 mRNA in Raw264.7 cells (P < 0.05). Co-cultured KCOT fibroblasts (KF) and HIOECs, respectively, expressed more COX-2 mRNA than the co-cultured gingival fibroblasts (GF) and HIOECs (P < 0.05). While the ratio of RANKL/OPG in HIOECs co-cultured with KF was also significantly higher than that co-cultured with GF (P < 0.05). The anti-human sRANKL antibody in KE-CM inhibited osteoclastogenesis of Raw264.7 cells; however, NS398 displayed little inhibition. CONCLUSION: An interesting phenomenon of osteoclastogenic effect of KE-CM in vitro was investigated, which suggested an indispensable role of epithelial-mesenchymal interaction of KCOT in its bone destruction. It could be at least partly attributed to the up-regulated ratio of RANKL/OPG in epithelium induced by KCOT fibroblasts, the aggressiveness of tumor as result of epithelial-mesenchymal interaction deserves exploration further.

Isolation and characterization of 32 microsatellite loci for topmouth culter (Culter alburnus Basilewsky).

The topmouth culter (Culter alburnus) is an economically important freshwater fish, which is widely distributed throughout large rivers, reservoirs, and lake areas of China. We report here the isolation and characterization of 32 new polymorphic microsatellite loci isolated from genomic DNA in this species enriched by (CA)12 and (GA)12 probes. The variability of these microsatellites was tested on 30 individuals cultured. The average allele number was 6.6 per locus, ranging from 3 to 12. The observed heterozygosity was from 0.4667 to 0.9000, and the expected heterozygosity was from 0.6163 to 0.9085. After using Bonferroni's correction for multiple tests, there was no evidence of linkage disequilibrium between pairs of loci, but deviations from Hardy-Weinberg equilibrium were found in 3 loci. These microsatellites can be used to study QTL of economic importance, population genetic diversity and the construction of genetic maps for C. alburnus in the future.

Regulatory functions of docosahexaenoic acid on ion channels in rat ventricular myocytes.

OBJECTIVE: The aim of this study was to study the regulatory functions of docosahexaenoic acid (DHA) on resting potential (RP), action potential duration (APD), delayed rectifier potassium current (Ik), and inwardly rectifier potassium current (Ik1) in rat ventricular myocytes, and analyze the related anti-arrhythmia mechanism. MATERIALS AND METHODS: Rat ventricular myocytes were isolated by enzyme digestion method. RP, APD, Ik and Ik1 in individual ventricular myocytes were recorded by patch-clamp technique with whole-cell configuration. Effects of DHA with various concentrations (0, 20, 40, 60, 80, 100 and 120 mmol/L, respectively) on RP, AP, Ik and Ik1 were investigated. RESULTS: There was no statistical difference of RP with different DHA concentrations (p > 0.05, n = 20), and the 25%, 50% and 90% of APD (APD25, APD50, and APD90) were gradually prolonged with increase of DHA concentration, respectively (p < 0.05, n = 20). IK gradually blocked and the I-V curve was downward shifted, according to increase of DHA concentration (p < 0.05, n = 20). The DHA half effect concentration (EC50) was 47.52 ± 2.32 µmol/L. With increasing DHA concentration, the steady-state inactivation curve shifted to left, and the recovery curve shifted to right. DHA had no significant effect on IK1 (p > 0.05, n = 20). CONCLUSIONS: DHA has regulatory functions on RP, APD, Ik and Ik1 in rat ventricular myocytes, which may be one of the related antiarrhythmic mechanisms.

[Clinical effects of ulnar artery perforator chain flaps in repairing wounds on distal forearm or wrist with vascular anastomosis].

Objective: To investigate the clinical effects of ulnar artery perforator chain flaps in repairing wounds on distal forearm or wrist with vascular anastomosis. Methods: The retrospective observational study method was used. From March 2015 to July 2019, a total of 11 serious trauma patients on distal forearm or wrist with vascular injury were admitted to the Second Hospital of Tangshan, including 8 male patients and 3 female patients, aged from 25 to 62 years, with an average age of 45 years. Vascular anastomosis and tendon repair were performed in all patients, and fracture reduction and fixation were conducted in 9 patients, of which 8 patients underwent external fixation of radius. The residual wounds were located in palmar distal forearm in 5 patients, palmar wrist in 4 patients, dorsal distal forearm in 1 patient, and dorsal wrist in 1 patient, with wound sizes ranged from 4.5 cm×3.0 cm to 10.0 cm×6.0 cm after the last debridement. The wounds were repaired with ulnar artery perforator chain flaps, with the flap sizes ranged from 5.2 cm×3.5 cm to 11.0 cm×7.0 cm. The wound in flap donor site of 1 patient was sutured directly, the wounds in flap donor sites of the other 10 patients were repaired with free skin grafts from ipsilateral thigh after being sutured partially, and the sizes of free skin grafts ranged from 4.0 cm×2.0 cm to 8.5 cm×5.0 cm. The survivals of flaps, skin grafts, and injured limbs after operation were observed. The appearances of the flaps and donor sites of flaps were observed during follow-up. At the final follow-up, the static two-point discrimination distances of the flaps were measured, and the satisfaction degrees of patients for the appearances of injured limbs were evaluated based on Michigan Hand Function Questionnaire. Results: All flaps, skin grafts, and injured limbs survived after operation, without wound infection and blister formation. All patients were followed up for 8 to 26 months, the appearances of the flaps were good and not bloated, with similar color, texture, and thickness to the surrounding skin. The donor sites of flaps repaired with skin grafts were smooth, with circle scar at the edges. At the final follow-up, the static two-point discrimination distances of the flaps was 10-15 mm; 7 patients were strongly satisfied with the appearances of the injured limbs, and the remaining 4 patients were satisfied with the appearances of flaps. Conclusions: The ulnar artery perforator chain flap has constant vascular anatomy and reliable blood supply, with simple operation, which provides a good treatment method for repairing wounds on distal forearm or wrist with vascular anastomosis. It is especially suitable for the patients with radius fracture fixed by external fixator.

Mutations in the beta-myosin heavy chain gene in southern Chinese families with hypertrophic cardiomyopathy.

In this study, 14 unrelated hypertrophic cardiomyopathy (HCM) probands were scanned by polymerase chain reaction-single-strand conformation polymorphism analysis and DNA sequencing. Three mis-sense mutations of the beta-myosin heavy chain gene, MYH7, were found: valine (Val) 606 methionine (Met), arginine (Arg) 694 leucine (Leu), and Arg 723 glycine (Gly). All are reported here for the first time in Chinese subjects. The results showed that: Val606Met is an intermediate malignancy mutation; Arg694Leu is a novel mutation with a benign phenotype; and the Arg723Gly mutation is linked to malignancy - it can lead not only to HCM but also to dilated cardiomyopathy at various ages. The clinical symptoms associated with Arg723Gly emerged early and caused more severe clinical manifestation and poorer prognosis in females than in males. Mis-sense mutations were not detected in the myosin binding protein C, cardiac, cardiac troponin T type 2, or cardiac troponin I type 3 genes. The MYH7 gene may be an HCM mutation hotspot in the Chinese and have unique features in this study population.

Intestinal crypt stem cells possess high levels of cytoskeletal-associated phosphotyrosine-containing proteins and tyrosine kinase activity relative to differentiated enterocytes.

Growth and differentiation of stem cells is thought to be regulated by growth factors and responding protein tyrosine kinase activities. Comparing mitotic stem cells from the adult intestinal epithelium, isolated from the crypts of Lieberkuhn, with isolated differentiated absorbtive cells we find major differences in the levels of phosphotyrosine-containing proteins. Crypt stem cells possess two major phosphotyrosine-containing polypeptides of 36 and 17 kD which have greater than 15 times more phosphotyrosine than that present in the polypeptides of differentiated enterocytes. Tyrosine kinase activity and similar phosphotyrosine-containing proteins are associated with the Triton cytoskeleton. Moreover, crypt tyrosine kinase(s) is active in vitro in phosphorylating similar cytoskeleton-associated substrates. These results suggest that cytoskeleton-associated phosphotyrosine kinase(s) and their substrates may play a role in growth and differentiation of adult intestinal epithelial cells.

Phorbol ester treatment stimulates tyrosine phosphorylation of a sea urchin egg cortex protein.

Fertilization of the sea urchin egg results in the phosphorylation, on tyrosine, of a high molecular weight protein localized in the egg cortex. In the present study, treatment of unfertilized eggs with the phorbol ester 12-O-tetradecanoylphorbol 13-acetate stimulated tyrosine phosphorylation of the high molecular weight cortical protein to levels three- to fivefold higher than that occurring in response to fertilization. Experiments using agents that inhibit the egg Na+/H+ exchange system or mimic the fertilization-induced shift in cytoplasmic pHi, suggest a signal transduction pathway in which protein kinase C activates the egg Na+/H+ exchange system and the resultant cytoplasmic pHi shift promotes tyrosine phosphorylation of the high molecular weight cortical protein.

Mesenchymal stem cells as a gene delivery system to create biological pacemaker cells in vitro.

Pacemaker cells differ from common cardiomyocytes due to the presence of a spontaneous depolarization process during the diastolic phase of the cardiac cycle. This is due to hyperpolarization-activated cyclic nucleotide-gated (HCN) channels, which are responsible for providing an inward current. Genetically engineered mesenchymal stem cells (MSCs) were transfected with hHCN4 genes using lentiviral transfection, and their potential use as biological pacemaker cells was investigated. In addition to expressing an anticipated high level of the hHCN4 gene, MSCs transfected with hHCN4 genes also expressed characteristic hHCN4 protein, a cardiac pacemaker-like current and were capable of increasing the spontaneous beating rate of co-cultured cardiac myocytes. Control MSCs did not exert these effects. It is hypothesized that genetically engineered MSCs transfected with hHCN4 genes by lentiviral transfection can be modified to be cardiac pacemaker cells in vitro.

Clinicopathogenic studies of acute diarrhea in children.

AIM: To identify etiologic pathogens of acute diarrhea in children and to determine the diagnostic value of stool pH. METHODS: From May 1988 to April 1992, 368 children with acute diarrhea were studied. Fresh stools were routinely examined, and stool pH was tested with pH paper. Samples were placed in Cary-Blair culture medium and were sent to the lab for bacterial isolation and identification. Rotavirus was identified in the supernatant by ELISA. RESULTS: Thirty-one pathogens and 385 bacterial strains were found in the 368 samples, with a detection rate of 67.7%, including 37.8% of mixed infections. Among the bacteria families, vibrionaceae was the most common (39.7%), and among bacteria genera, aeromonas was the most common (26.8%). In bacterial diarrhea, stool pH tended to be basic, while in viral diarrhea it tended to be acidic. CONCLUSION: There are 31 pathogens for children's acute diarrhea in this area. It is quite difficult to make an etiologic diagnosis only by clinical signs. However, stool pH is of some value for early disease diagnosis.

[Insight into the mechanism of torsade de pointes (TDP) induced by cesium chloride].

Standard intracellular potential recording technique was used in isolated guinea pig ventricular muscle and monophasic action potential (MAP) recording technique was performed on both the epicardium and endocardium of dog hearts in vivo. The results showed that CsCl lengthened action potential durations of ventricular muscle as well as QT intervals of dog hearts. The incidence of early after depolarization (EAD) induction was 71.4%. Ventricular tachycardia induced by CsCl manifested as typical TdP in 44.4% with EAD shown on epicardial MAP recording. This suggested that EAD and triggered activity might be an important mechanism of TdP. As verapamil could depress the induction of EAD, we believe that Ca++ inward currents was responsible for the induction of EAD and TdP by CsCl.

[Use of signal averaging system in the study of ventricular late potentials].

Signal-averaged electrocardiograms (SAECGs) were performed in 93 patients with various heart diseases and 20 dogs with experimental acute myocardial infarction. SA-ECGs were recorded with a high-pass bidirectional digital filter Mode FDS which was made by Shanghai Fudan University and Suzhou Medical College. All the patients and dogs were also examined with ART 1200 EPX recorder. Ventricular late potentials (VLPs) were directly recorded from the epicardium with experimental infarction by using composite electrodes and signal averaging system in late stage of coronary ligation. The results showed: VLPs were detected by body surface SA-ECG only in half of the dogs with VLPs detected by the real time mapping on infarction region, but the abnormal SAECGs did reveal the delayed ventricular activity; the data obtained with FDS or ART SA-ECG were similar.

[Electrophysiologic studies on reentrant ventricular arrhythmia in the late myocardial infarction period].

The mechanisms of reentrant ventricular tachycardia were studied in canine post-infarction model. In 15 dogs that weighed 10-20 kg, the anterior descending coronary artery was ligated. 3 to 7 days after coronary artery occlusion, the dogs were reoperated to set up a composite electrode or a patch electrode with 32 bipolar electrodes. The composite electrode was utilized to record ventricular late potentials (VLPs) from the epicardial surface of the infarction zone (IZ) and adjacent normal zone (NZ). The bipolar electrodes were used to record the epicardial isochronal maps. 16 bipolar electrodes were placed over the area of infarction and its bordering zone and the other 16 bipolar electrodes were distributed over the remaining surface of the ventricle. Reentrant ventricular tachycardias were induced by programmed electrical stimulation. The VLPs and epicardial isochronal activation were recorded by SC-16 oscillograph and computerized mapping system during sinus rhythm and programmed stimulation. The experiments showed; 1) VLPs were demonstrated in 11 of the 15 dogs (73.3%) during programmed stimulation. 2) The reentrant circuit has a characteristic figure-8 configuration in the form of two circulating wave-fronts around arcs of functional conduction block that coalesce into a slow commun reentrant wave-front. One of the two synchronous circuits traveled clockwise and the other counter-clockwise. 3) VLPs represent a delayed depolarization at the ischemic myocardium, where the impulse conducts slowly in the reentrant circuit. It is clear that the present study may increase the understanding of the mechanism of ventricular arrhythmias in the late myocardial infarction period.

[Studies on electrophysiology of lysophosphatidylcholine and evaluation of its clinical significance].

The cellular electrophysiological effects of lysosphatidylcholine (LPC) were investigated by using patch-clamp whole cell recording and conventional microelectrode technique, LPC(10 microns) suppressed the sodium, background potassium and calcium current and the effect of LPC on activity of ionic channels was nonselective. Resting potential, action potential amplitude and maximal rate of rise of phase of action potential were decreased with LPC (50 microns) perfusing. The incidence of abnormal automaticity, early after depolarization (EAD) and delayed after depolarization (DAD) was higher. It is clearly shown that LPC accumulates at early time of myocardial ischemia and thus may induce ischemic arrhythmias.

[A study on diarrhoea disease caused by Vibrionaceae along coast the east of Zhejiang Province].

A study of the etiology of 1028 cases of acute diarrhoea due to vibrionaceae was carried out between August 1988 to July 1990 along the coast of east Zhejiang Province. From 417 clinical cases (40.56%) were found. 590 strains (57.39%) of pathogenic vibrios Among them, the detection rates of vibrio, aeromonas and plesionmonas were 48.14%, 44.74%, 7.12% respectively. There were 8 Species of vibrio detected: V. parahemolyticus (63.73%), V. fluvialis (11.97%), V. alginolyticus (10.215), non-01 vibrio cholera (9.86%), V. furnissii (2.47%), V. mimicus (1.06%), V. metchnikovii (0.35%), V. vulnificus (0.35%). The detection rate of aeromonas species of A. sorbia, A. caviae, A. hydrophila and A. indefinite sp. were 30.68%, 27.27%, 22.73%, and 19.32% respectively. Plesionmonas shigellides was also found in this study. The age, sex, clinical features, detection rates of pathogenic vibrios of diarrhoea cases and sea food contact history were analyzed.

Using an abnormal increase in postexercise systolic blood pressure to diagnose coronary artery disease in gerontal patients.

Data from 66 patients ≥ 60 years old with suspected coronary artery disease (CAD) were studied to determine the diagnostic value of an abnormal increase in postexercise systolic blood pressure (SBP) for detecting CAD in gerontal patients. Treadmill exercise testing (TET) and selective coronary angiography (CAG) were carried out and SBP was measured pre-TET and at each minute during a 6-min post-TET recovery phase. Abnormal increase in postexercise SBP was defined as a higher SBP compared with that measured earlier during the 6-min post-TET period. An abnormal increase of ≥ 7 mmHg in postexercise SBP had a statistically significantly better specificity, and also showed higher sensitivity and accuracy, than ST-segment depression ≥ 1 mV in identifying gerontal patients with CAD. The combination of ST-segment depression and abnormal SBP resulted in further improvement of the specificity for detecting CAD. It is concluded that measurement of abnormal increase in postexercise SBP may be a sensitive indicator of gerontal CAD.