[Effects of low position lateral supramalleolar flap carrying periosteum and proximal leg propeller flap in relay repair of electric burn wounds of forefoot].

Objective: To explore the effects of low position lateral supramalleolar flap carrying periosteum and proximal leg propeller flap in relay repair of electric burn wounds of forefoot. Methods: A retrospective observational study was conducted. From January 2019 to January 2022, 12 patients with electric burn wounds of forefoot meeting the inclusion criteria were admitted to the Sixth Hospital of Shanxi Medical University, including 10 males and 2 females, aged 23-65 years. After debridement, the wound with an area of 6.0 cm×3.0 cm to 15.0 cm×7.0 cm was repaired with the lateral supramalleolar flap carrying part of the periosteum of the distal tibia and fibula with the rotation point moved down to the front of the ankle joint. The area of the cutted flap was 6.5 cm×3.5 cm-15.5 cm×7.5 cm. At the same stage, the donor site wound of lateral supramalleolar flap was repaired with peroneal artery or superficial peroneal artery perforator propeller flap in relay, with the relay flap area of 3.0 cm×1.5 cm-15.0 cm×4.0 cm. After operation, the survival of the lateral supramalleolar flap and relay flap, and the wound healing of the relay flap donor site were observed. During follow-up, the shapes of the lateral supramalleolar flap and its donor site were observed. Results: After operation, one patient developed secondary blisters in the superficial skin distal to the lateral supramalleolar flap, which healed after dressing change, and the lateral supramalleolar flap and relay flaps survived well in the other patients; the donor site wound of the relay flap healed well. During follow-up of 12-18 months, the lateral supramalleolar flaps were in good shape and not bloated, with only linear scar left in the donor site of the flap. Conclusions: The low position lateral supramalleolar flap carrying periosteum can repair electric burn wounds of forefoot with advantages including reliable blood supply, low rotation point, and better repair effects. The use of relay flap to repair the donor site of lateral supramalleolar flap can reduce the damage to the appearance and function of the donor site.

One-staged reconstruction of Achilles tendon and overlying skin defects with suppuration: using peroneus brevis tendon transfer and reversed sural neurofasciocutaneous flap.

The combined loss of the Achilles tendon and the overlying soft tissue with suppuration has been treated with many single- or multi-staged procedures, most of which are technically complex. The ideal single-stage procedures are needed. Ten patients with combined loss of the Achilles tendon and the overlying soft tissue underwent reconstruction using peroneus brevis tendon transfer and reversed sural neurofasciocutaneous flap. Follow-up was 8-48 months. Of the 10 flaps, 9 survived uneventfully except for 1 flap that had distal marginal necrosis; the flap healed following a dressing change. The flaps were wear-resistant and cosmetically acceptable. All the patients were able to perform heel lift with the operated limb and resumed walking. No Achilles tendon re-rupture or misbalance of ankle joints occurred as of date. The Arner-Lindholm evaluation standard was taken to evaluate the curative effect; the results were excellent in seven cases and good in three cases. As a one-stage procedure, the peroneus brevis tendon transfer and reversed sural neurofasciocutaneous flap is an ideal option to reconstruct the combined loss of the Achilles tendon and the overlying soft tissue.

Characterization of some biochemical properties of bradykinin-induced cAMP and cGMP formation in guinea pig ileum.

One of the receptor-mediated events, cyclic nucleotide, i.e. cAMP and cGMP formation induced by bradykinin in guinea pig ileum was investigated in this report. Bradykinin, similar to acetylcholine, produced a rapid rise in the levels of cAMP and cGMP in the ileum. The absolute amount of these cyclic nucleotides induced by the same dosages (10(-8) to 10(-6) M) of bradykinin was greater in cAMP than in cGMP. This increase in cyclic nucleotides was dependent upon the presence of calcium in the medium. Addition of EGTA (0.1 mM) in a calcium free medium resulted in a significant reduction of the levels. The elevation of cAMP and cGMP levels induced by bradykinin in the ileum could not be blocked by either atropine (an anticholinergic agent) or propranolol (a beta-adrenergic blocker). Both of these blockers did not alter the basal levels of two cyclic nucleotides. However bradykinin-induced cAMP formation could be completely blocked by either indomethacin (a prostaglandin synthesis inhibitor) or dexamethasone (a phospholipase A2 inhibitor), This, however, was not true in the case of bradykinin induced cGMP formation. Additionally, both blockers did not create a significant change in the basal levels of these cyclic nucleotides. Bradykinin induced cAMP formation in the ileum was indicated by observed results to likely occur through an indirect process, i.e. the formation and release of prostaglandin in the cell, whereas bradykinin-induced cGMP formation did not. The elevation of these cyclic nucleotides in the cells was observed to be related to the movement of calcium ion across the cell membrane.

Characterization of 3H-serotonin (5-HT) binding and effects on the phosphoinositides (PI) turnover in cultured C6 glioma and N2 neuroblastoma cells from rodents.

3H-5-HT (serotonin) binding and its displacement by various specific subtype ligands and effects on the phosphoinositides (PI) turnover were studied in cultured C6 glioma and N2 neuroblastoma cells from rodents. Saturation analysis of 3H-5-HT binding to C6 cells revealed that its Kd and Bmax were 3.0 nM and 18.0 pmole/mg protein respectively. DOI.HCl (5-HT2 agonist) and ketanserin (5-HT2 antagonist) had the highest affinities in the drug-displacement of 3H-5-HT binding to C6 cells studied. The IC50 values for DOI-HCl and ketanserin were 7.5 x 10(-7) and 3.5 x 10(-8) M respectively. 5-HT also induced 3H-PI breakdown and generated 3H-IP. The EC50 values for 5-HT for this event were in the dose range between 0.5 to 1.5 microM, and this 5-HT-induced response could be blocked by 5-HT2 antagonist ketanserin more effectively than the 5-HT1 antagonist or 5-HT3 antagonist studied. 3H-5-HT binding to N2 cells revealed that its Kd and Bmax were 4.0 nM and 80 pmole/mg protein respectively in the saturation analysis study. The drug-displacement of this binding revealed that MDL 72222 (5-HT3 antagonist) had a higher affinity than ketanserin. The IC50 values for MDL 72222 and ketanserin were 10 nM and 10 microM respectively, when 3 nM of 3H-5-HT was used. Our results indicate that the predominant receptor subtype of 5-HT in C6 and N2 cells are 5-HT2, and 5-HT3 respectively, and that the PI turnover is linked to 5-HT2, but not 5-HT3 receptor activation.

Effects of insulin on glucose uptake in cultured cells from the central nervous system of rodent.

1. Incubation of C6 glioma cultures with insulin resulted in a time and dose-dependent stimulation of 2-deoxy-D-glucose uptake. The maximal stimulation (160% of the control) was observed with 1 nM insulin and 0.05 nM caused half-maximum effect. 2. Incubation of NG 108-15 (neuroblastoma x glioma hybrid) and N2 neuroblastoma cells with 160 nM insulin did not result in a significant stimulation of this glucose uptake. 3. The basal level and stimulatory effect by insulin on this glucose uptake observed in C6 glioma cells were dependent on the presence of calcium in the medium. 4. Such an increase in glucose uptake in C6 glioma cells was also observed in the presence of diacylglycerol (DG) generating agents, such as carbachol (1 mM) and phospholipase C (0.05 unit/ml) or of DG analogs, such as sn-1,2-dioctanoyl glycerol (250 microM) and phorbol myristate acetate (1 microM). 5. Our results indicated that both calcium ion and DG levels play important roles in the regulation of glucose uptake in the glial cells, but not in neuronal cells from the brain.

Evidence for the coupling of muscarinic M3 receptor to cyclic AMP formation and poly-phosphatidylinositol turnover in rat salivary glands.

The biochemical events which are coupled to the subtypes of muscarinic cholinergic receptors in rat salivary glands were studied. The subtype property of this receptor system was characterized by the use of 3H-QNB binding to glandular membrane homogenates. The Kd and Bmax values of this binding were found to be 0.2 nM and 210 fmole/mg protein respectively. In the drug-displacement study on the 3H-QNB binding the following potency order was obtained (based on the IC50 values calculated from each individual dose-response curve): Atropine greater than 4-DAMP greater than HHSiD greater than Pirenzepine greater than AF-DX 116. This order is a typical M3 muscarinic subtype, according to the recent consensus. Carbachol (0.1 mM) caused a 4.4-fold increased in IP3 production over the basal level, when tissue fragments were prelabeled with 3H-inositol. The above mentioned cholinergic antagonists could block this event in a similar potency order. Carbachol (0.1 mM) did not have a significant effect on the basal level of cAMP formation of these tissue homogenates. On the other hand, it had a 33% reduction of isoproterenol (10 microM)-enhancing cAMP formation. The reduction effect of carbachol on cAMP formation could also be blocked by the above mentioned cholinergic antagonists in a similar order. Our results indicated that rat salivary glands have M3 muscarinic receptors and the activation of these receptors causes changes in both phosphatidylinositol turnover and cAMP formation.

Biochemical events coupled to the activation of serotonin receptors in brain regions of the rat.

Effects of serotonin on the levels of inositol phosphate and cyclic AMP in certain brain regions of the rat were studied. When brain slices were treated with serotonin (10(-4) or 10(-5) M), it caused significant increases in inositol phosphate levels in cerebral cortex, hippocampus, midbrain and brain stem, but not in cerebellum. Serotonin itself did not have a significant effect on the basal levels of cAMP in most of these regions, except in midbrain in which it had stimulatory effect. However, serotonin also had significant inhibitory effect on forskolin-induced cAMP formation in cerebral cortex, hippocampus and brain stem but not in midbrain in which an increase instead. Effects of serotonin on both biochemical events were in dose-dependent manner, and their maximal responses were at doses between 10(-4) and 10(-3) M. The rank order of the maximal response to serotonin on inositol phosphate production in these regions was cerebral cortex greater than hippocampus greater than midbrain greater than brain stem greater than cerebellum. On the other hand, the rank order of the maximal response to serotonin on inhibiting forskolin-induced cAMP formation was cerebral cortex = brain stem greater than hippocampus. Our data revealed that there is a regional difference in serotonin-induced inositol phosphate and cAMP responses. This difference is likely due to a differential distribution of serotonergic receptor subtypes and densities in different regions of rat brain.

[Ultrasonic diagnosis of intraocular cysticercus].

Intraocular cysticerci are mostly located in the subretinal space or vitreous. When the fundus is obscured by exudate or hemorrhage, the diagnosis becomes very difficult with the ophthalmoscope or biomicroscope. The authors performed A- and B-scan echographic studies on 20 cases of intraocular cysticercus during February 1985 to December 1988; it was found that B-scan was very useful in diagnosing intraocular cysticercus with regard to its location and relation to surrounding tissues. Several cases with typical echograms are presented with discussions. The ultrasonic diagnoses were confirmed by surgery.

Guanine nucleotides have a direct inhibitory effect on polyphosphoinositide turnover in rat cortical synaptosomes.

1. The possible involvement of guanosine 5'-triphosphate (GTP)-binding proteins in the receptor mediated polyphosphoinositide (PPI) turnover event was investigated in rat cortical synaptosomes. 2. It was studied under the effects of guanine nucleotides on 32Pi incorporation into synaptosomal phospholipids in the absence or presence of carbachol. 3. The basal 32Pi incorporation into these phospholipids was altered by the presence of 1 mM carbachol: i.e. a decrease in 32Pi incorporation into phosphatidylinositol-4,5-bisphosphate and phosphatidylinositol-4-phosphate and an increase in the incorporation of 32Pi into phosphatidylinositol and phosphatidic acid. 4. In the presence of guanine nucleotides: GTP, Gpp(NH)p and GDP at suitable concentrations, there was a general decreasing effect on 32Pi incorporation into all 4 phospholipids, which are all involved in PPI turnover cycle, either in the basal or carbachol-stimulated levels. 5. There was no selective effect among the guanine nucleotides studied on this PPI turnover event. It is, therefore, likely that these nucleotides have a direct inhibitory effect on PPI turnover, and this action may not act through a GTP-binding protein.

Evidence for the coupling of muscarinic M1 receptors to polyphosphoinositide turnover in rat cortical synaptosomes.

The linking event between the activation of muscarinic M1 receptors and the stimulation of polyphosphoinositide (PPI) turnover was studied in rat brain synaptosomes from the muscarinic M1 enriched cerebrum and M2 enriched cerebellum. The muscarinic M1 selective antagonists, pirenzepine and trihexyphenidyl, and M2 selective antagonist AF-DX-116 were chosen to displace the non-selective labeled cholinergic ligand, 3H-QNB, binding to these two regions of the brain synaptosomes. The IC50 values obtained for these agents revealed a typical characterization of the receptor subtypes of these two regions as they are. Carbachol-induced a stimulation of the PPI turnover cycle: namely, a decrease in 32Pi incorporation into phosphatidylinositol-4,5-bisphosphate (TPI) and phosphatidylinositol-4-phosphate (DPI), and an increase in this incorporation into phosphatidylinositol (PI) and phosphatidic acid (PA) in rat brain synaptosomes from the cerebrum. However, this event was only barely detectable in the synaptosomes from the cerebellum. The IC50 values obtained for these antagonists to block the carbachol-induced PPI turnover cycle in the synaptosomes from the cerebrum were close to the values obtained for the displacement of 3H-QNB binding to the same preparation, and were far away from those values obtained in the synaptosomes from the cerebellum. Our results suggest that there is evidence to support the view that muscarinic M1 receptors are coupled to the PPI turnover event in rat cortical synaptosomes.

Studies of blocking mechanisms of carbachol-induced polyphosphoinositide turnover in rat cortical synaptosomes by neuroactive drugs.

1. The incorporation of 32Pi into 4 phospholipids of rat cortical synaptosomes was altered in the presence of carbachol (1 mM), viz. a decrease of phosphatidylinositol-4,5-bisphosphate and phosphatidyl-4-phosphate by 34 and 21%, and an increase of phosphatidylinositol and phosphatidic acid by 52 and 96% of basal controls respectively. 2. The IC30 values calculated from the dose-response curves for drugs affecting carbachol-induced 32Pi incorporation into these phospholipids, and [3H]QNB binding to the cortical synaptosomes were similar for the typical antimuscarinic agents (i.e. atropine, pirenzepine and trihexyphenidyl), and tricyclic antidepressants (i.e. amitriptyline, doxepin and imipramine) studied. 3. The IC30 values obtained for drugs affecting carbachol-induced 32Pi incorporation into these phospholipids, and high potassium-induced 45Ca2+-uptake by this preparation were similar for neuroselective calcium channel blockers (i.e. cinnarizine and flunarizine) studied. 4. Our results suggest that the neuroactive drugs studied can either act at, or beyond the receptor level, perhaps on the availability of calcium ion, to block carbachol-induced polyphosphoinositide turnover in rat cortical synaptosomes.

Effects of calcium antagonists on (Na+ + K+)-ATPase, Mg2+-ATPase and Ca2+-ATPase activities of rat cortical synaptosomes.

1. The effects of 11 calcium antagonists on (Na+ + K+)-ATPase, Mg2+-ATPase and Ca2+-ATPase activities of rat cortical synaptosomes were studied. 2. All the calcium antagonists studied had inhibitory effects on ouabain-sensitive (Na+ + K+)-ATPase, Mg2+-ATPase and Ca2+-ATPase activities in synaptosomes at high concentrations (10 or 100 microM). 3. Calcium antagonists such as trifluoperazine, flunarizine and cinnarizine had inhibitory effects on Ca2+-ATPase activity at low concentrations (1-10 microM). 4. Trifluoperazine and La3+ had inhibitory effects on Mg2+-ATPase activity at low concentration (1 microM). 5. Our results suggest that most of the calcium antagonists studied have little effects on neuronal (Na+ + K+)-ATPase, Mg2+-ATPase and Ca2+-ATPase activities at therapeutic dose ranges (1 microM or lower).

Effects of anticonvulsant drugs on calcium transport and polyphosphoinositide metabolism in rat cortical synaptosomes.

1. Anticonvulsants: phenytoin, phenobarbital, carbamazepine and valium at concentrations of 10-100 microM had a significantly inhibitory effect on both K+-stimulated Ca2+-uptake and 32Pi incorporation into phospholipids of rat cortical synaptosomes. 2. Other anticonvulsant, valproic acid, at concentration upto 100 microM had no effect on these two events. 3. Our results suggest that there is a link between Ca2+-influx and polyphosphoinositide turnover in synaptosomes, and this link may relate to the inhibitory effect of these drugs on neurotransmitter release mechanisms of this preparation.

Effects of calcium ionophore A23187 and calcium antagonists on 32Pi incorporation into polyphosphoinositides of rat cortical synaptosomes.

The role of Ca2+ on 32Pi incorporation into polyphosphoinositides (PPI) of rat cortical synaptosomes was studied. Stimulation of muscarinic receptor by carbachol (1 mM) resulted in a decrease in 32Pi incorporation into phosphatidylinositol-4,5-bisphophaphate (TPI) and phosphatidylinositol-4-phosphate (DPI), and an increase in 32Pi incorporation into phosphatidylinositol (PI) and phosphatidic acid (PA), whereas no significant effect on other membrane phospholipids was found. This response could be blocked by atropine (1 microM). The stimulatory effect of carbachol required Ca2+ in the medium; the presence of 0.5 mM EGTA blocked the effect of carbachol on PPI turnover completely. Calcium ionophore A23187, at 1 microM, had a similar effect on PPI turnover by carbachol (1 mM). At higher concentrations (10-100 microM) of A23187, the PPI turnover rate was much enhanced. Depolarization of the membrane by high potassium (60 mM) in the presence of calcium resulted in an enhanced PPI turnover, which was similar to the results of the carbachol (1 mM) effect but to a lesser extent. Calcium antagonists, diltiazem and trifluoperazine, at 10 microM could block the carbachol effect on 32Pi incorporation into PPI in this preparation. Our results suggest that the enhancement of PPI turnover in rat cortical synaptosomes by carbachol, calcium ionophore or high potassium requires Ca2+, and it can be blocked by compounds which interfere with the availability of this ion, such as EGTA or calcium antagonists.

Cations and calmodulin in normal and neoplastic cell growth regulation.

The purpose of this presentation is to review pertinent literature pertaining to the role of divalent cations and calmodulin in regulating growth of nonneoplastic and neoplastic cells and to examine the anticancer efficacy of some calmodulin inhibitors. Although normal eukaryotic cell replication and proliferation is closely controlled by a complex system of endogenous substances, it is likely that the coordination of purposeful interactions between these substances is the ultimate responsibility of two groups of cellular components, namely the divalent cations Ca2+ and Mg2+ and the versatile intracellular Ca2+-binding protein calmodulin (CaM). When free Ca2+ enters normal cells, it acts as a positive signal for proliferation; this action appears to be specifically associated with the late G1 phase, just prior to DNA synthesis. This period is designated G1/S and is considered to contain Pardee's "restriction point." Reduction of extracellular Ca2+ concentrations between physiological levels (1-0.1 mM) results in gradually reduced rates of cell proliferation; at Ca2+ concentrations of 0.1 mM or less, normal cell proliferation is reversibly inhibited. Since an extracellular concentration of about 0.7 mM Mg2+ is required for Ca2+ to initiate cell replication, it has been proposed that Ca2+ and Mg2+ act in concert via a common mechanism, however, in contrast to Ca2+, Mg2+ appears to be required throughout the entire cell cycle. Intracellular Ca2+ can activate CaM which, in turn, can modulate various cellular processes that affect cell proliferation, including cyclic nucleotide metabolism, protein phosphorylation, polyamine metabolism, prostaglandin metabolism, Ca2+ transport, DNA synthesis, and microtubular function including mitosis.(ABSTRACT TRUNCATED AT 250 WORDS)

Inhibition of human breast cancer colony formation by anticalmodulin agents: trifluoperazine, W-7, and W-13.

The effects of anticalmodulin agents, namely trifluoperazine (TFP) and two naphthalene sulfonamide derivatives (W-7 and W-13), were tested on the growth of a human breast cancer cell line (MDA-MB-231) using a soft agar clonogenic assay. The results of this in vitro study reveal that TFP, W-7, and W-13 had the ability to inhibit the colony formation from this cell line. The inhibitory effect was greater when the cancer cells were exposed to these agents continuously than when the cells were exposed to the drugs for 1 h. The IC50 values for TFP, W-7, and W-13 in continuous exposure were about 18, 30, and 38 microM, respectively, whereas the corresponding values for 1-h exposure were 50, 53, and 70 microM, respectively. These findings suggest that anticalmodulin agents can inhibit the growth of human cancer cells at relatively low concentrations in vitro. Whether effective antitumor concentrations of these drugs can be achieved in vivo remains a subject for further study.

Decreased activities of cyclic cytidine 3',5'-monophosphate phosphodiesterase in Morris hepatomas having varying growth rates.

1. The activities of cyclic cytidine 3',5'-monophosphate (cCMP) phosphodiesterase in normal rat liver and host liver (bearing hepatoma 5123 t.c.(h)) were compared with those of three Morris hepatomas of varying growth rates. 2. The results show that the order of enzyme activity was as follows: normal liver = host liver greater than 7794A (slow growth rate) greater than 5123 t.c.(h) (intermediate growth rate) greater than 7800 (fast growth rate). 3. The enzyme had a pH optimal value of about 7.0 and an apparent Km for cCMP about 2.8 mM; its activity was slightly affected by the presence of calmodulin (100 micrograms/ml) and/or CaCl2 (100 microM), but showed variable responses to other cations (La3+, Mg2+, Mn2+, Zn2+, Fe2+, Na+ and K+).

Positive correlation between calmodulin content and hepatoma growth rates.

Calmodulin contents of normal rat liver, host liver [bearing hepatoma 5123t.c.(h)], regenerating liver, and Morris hepatomas 7800, 5123t.c.(h), and 7794A were determined by phosphodiesterase assay and by radioimmunoassay. The calmodulin levels determined by both assays were significantly increased in three hepatomas when compared to the corresponding values of normal liver. The order of increase in calmodulin content was as follows: normal liver = host liver less than 7794A (slow growth rate) less than 5123t.c.(h) (intermediate growth rate) less than 7800 (fast growth rate). In regenerating liver (24 hr after partial hepatectomy), the calmodulin content was not different from that of normal liver. In good agreement with the literature, the calmodulin values measured by the phosphodiesterase assay were always lower than those determined by radioimmunoassay. Calcium and magnesium contents were measured by atomic absorption spectrophotometry in acid digests of these tissues. Both cation contents were significantly increased in the three hepatomas studied when compared to the corresponding values of normal liver; the extent of increase for calcium content (120 to 240%) was much greater than that for magnesium (30 to 40%). The order of increase for both cations was as follows: normal liver = host liver less than 5123t.c.(h) less than 7794A less than 7800. Therefore, there does not appear to be any correlation between the cation contents and hepatoma growth rates. In regenerating liver, magnesium content was about 14% higher than that of normal liver. In summary, the results indicate that only the increase of calmodulin appears to correlate positively with the growth rate of these tumors. This correlation suggests that calmodulin may be involved in tumor cell growth regulation.