[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.

The roles of MAPKs in rabbit nucleus pulposus cell apoptosis induced by high osmolality.

BACKGROUND: Earlier work has suggested that the p38 MAPK, JNK1/2 and ERK1/2 signal pathway existed in nucleus pulposus cells and the cell growth, differentiation and apoptosis were regulated by them. Because osmotic fluctuations are inevitable in the physicochemical environment of intervertebral disc cells, high osmolality could activate p38 MAPK, JNK1/2 and ERK1/2 signal pathway. The effects of high osmolality on the catabolic program and proliferation of nucleus pulposus cells are still not clear. AIM: To explore the possible roles of MAPKs in rabbit nucleus pulposus cell apoptosis induced by high osmolality. MATERIALS AND METHODS: Rabbit nucleus pulposus cells were cultured and divided into different group at random. The cells were pretreated with inhibitor for p38 MAPK, JNK1/2 and ERK1/2 signal pathway respectively. In next step, the cells were cultured in different osmolality environment for different time at 37°C in 5% carbon dioxide incubator. After treatments, ratio of apoptosis was measured by flow cytometry, and western blotting was performed to quantify the expression of the activated forms of p38 MAPK, JNK1/2 and ERK1/2. Furthermore, immunofluorescence analysis with confocal microscopy was performed to confirm the hyperosmolality effects on activation of p38 MAPK, JNK1/2 and ERK1/2 signal pathways in nucleus pulposus cells. RESULTS: Our results show that in 500 and 600 mOsm/kg medium, rabbit nucleus pulposus cell apoptosis increased, and a persistent phosphorylation of p38 MAPK, JNK1/2 and ERK1/2 proteins were observed. In the same condition, the apoptotic cells death remarkably decreased when the p38 MAPK and JNK1/2 signal pathways were blocked by their inhibitors SB203580, SP600125 repectively. On the other side, the apoptotic cells death rate reraised greatly when the ERK1/2 signal pathways were blocked by its inhibitor PD98059. CONCLUSIONS: High osmolality activated p38MAPK, JNK1/2 and ERK1/2 in rabbit nucleus pulposus cell, and the activated p38 MAPK and JNK1/2 induced cell apoptosis, on the contrary, the activated ERK1/2 made the cell survived.

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.

[Study on the analysis of alpha-naphthylacetic acid].

This paper introduces a method for the determination of alpha-naphthylacetic acid with HPLC. Under the reversed-phase condition, puting ion-pair reagent IPR-A in the mobile phase of methanol-water, an unknown peak was separated. The recovery was 100.2% and the RSD was 0.30%. In comparing with conventional HPLC using methanol-water as mobile phase and GC methods, the results were more dependable.

[Research on improving rice resistance to the pest by B.t. and SBTi genes].

B.t. gene alone or and SBTi gene together were introduced into two elite indica rice varieties grown in South China by bombardment. And 21 independent transgenic lines containing B.t. gene and 4 independent transgenic lines containing B.t. gene and SBTi gene were obtained. Molecular and genetics analysis for R1 plants showed integration of multiple transgenes occurred at one genetic locus. Northern blot result proved B.t. gene expressed in R2 transgenic plants stably. Bioassays using R2 transgenic plants with leaf-folder (Cnaphalocrosis medinalis), indicated that transgenic rice plants are more resistant to the pest than untransformed control plants. And those transgenic plants containing B.t. and SBTi genes showed more resistance compared with those plants containing B.t. gene.

[Fertile transgenic indica rice from microprojectile bombardment of embryogenic callus].

A reproducible and efficient transformation system for indica rice was developed based on microprojectile bombardment of embryogenic callus. Sufficient globular embryogenic calli were produced within 2-6 weeks from 3-4 week old calli derived from mature embryos. The embryogenic calli were bombarded with the plasmid pFWZ16 containing bar gene and B.t. delta-endotoxin gene. Selection, pre-regeneration, regeneration and rooting were carried out on media with 2-4 mg/L Basta. Partial desiccation treatment was used to increase the efficiency of regeneration. Transformed plantlets were recovered within 4-5 months after mature seeds were plated. From 821 bombarded embryogenic calli of two elite rice varieties popularized in South China, 477 Basta resistant plants of 48 lines were obtained. Results of PCR, Southern blotting, RNA dot blotting, and topical application of Basta demonstrated that the foreign bar gene and B.t. gene were integrated into the genome of transgenic rice plants, and also expressed. 87.5% of the transgenic plants were fertile. Mendelian segregation of the foreign genes was indicated by Basta application and Southern blot analysis of R1 plants.

Effects of ethanol on brain monoamine content of spontaneously hypertensive rats (SHR).

Spontaneously hypertensive rats (SHR) were administered either 2.4 g/kg ethanol or an isocaloric glucose daily for 4 weeks and the levels of norepinephrine (NE), epinephrine (EP), dopamine (DA), serotonin (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) in different brain regions were determined. Results indicated a 3-fold increase in NE level in brain stem and hypothalamus and more than 2-fold increase in DA in corpus striatum in alcohol-treated rats as compared to controls. There was a significant increase in the level of DA in the corpus striatum but the levels in cerebral cortex, brain stem and hippocampus were decreased instead. Decreases in 5-HT levels were found in hypothalamus, brain stem, cortex and cerebellum of alcohol-treated brain as compared to untreated controls. These results indicate alterations of the biogenic amine contents in different regions of the SHR brain after chronic ethanol ingestion. Since stimulated release of biogenic amines in the SHR brain has been implicated in the regulation of blood pressure, changes due to ethanol ingestion may be a risk factor in hypertensive patients.

Differential responses in cyclic GMP formation induced by excitatory amino acids (EAA) and sodium nitroprusside (SNP) in various regions of the brain and of rats of varied age.

1. Sodium nitroprusside (SNP, 100 microM) caused a rapid and great increase of formation of cGMP in rat cerebellar slices. This effect was not blocked by L-NMMA (a NO synthetase inhibitor) or antagonists of the NMDA receptor complex (e.g. AP5 or MK 801). 2. Similarly, NMDA (100 microM) and glutamate (1 mM) caused a rapid but less significant increase of cGMP formation. This increase was blocked by NMDA receptor complex blockers (e.g. AP5, MK801 and kynurenate), and L-NMMA and L-nitroarginine. 3. In rats aged 12 days, both NMDA and kainate (at 100 microM) caused significantly increased levels of cGMP in the cerebellum, pons and medulla areas, whereas no significant alterations were found in the cerebral cortex, hippocampus or midbrain areas. 4. NMDA (100 microM) and SNP (300 microM) induced greater increases of cGMP in cerebellar slices in young (aged 13 days) animals than older ones of either sex. This effect decreased greatly after 35 days of age. In adult (2 months) animals the effect of NMDA had virtually disappeared whereas SNP was barely significantly present. 5. Our results suggest that brain region and age, but not sex, affected formation of cGMP induced by excitatory amino acids (EAA) and SNP. Furthermore, endogenous NO production is required by EAA, but not by SNP, in the formation of cGMP.

[Changes in plasma interleukin-1 and their possible relationship with the changes in glucocorticoid receptor in aged long-distance runner].

For the study of the changes in plasma interleukin-1 (IL-1) and their possible relationship with the changes in glucocorticoid receptor (GR), plasma IL-1 and GR in peripheral blood leukocytes in aged long-distance runner were measured simultaneously. The activity of IL-1 was expressed as its ability to stimulate 3H-TdR incorporation in the thymocytes of C57 mice. GR was determined by whole cell assay with 3H-Dex. The results showed that the activity of plasma IL-1 in aged long-distance runner was 209%, 223% and 145% of the control at 14.7-18.7, 3.8-7.0 and 1.5-2.6 KD fractions. The GR in peripheral blood leukocytes in aged runner was 65% of the control. Possible relationship between the changes in IL-1 and GR in aged long-distance runner and its physiological significance are discussed.

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.

Studies on tachykinin (neurokinin) receptor coupled with inositol phospholipid hydrolysis and salivation in rat salivary glands.

3H-substance P binding to the membranes of rat salivary glands was studied. The Kd and Bmax values were found to be 0.34 nM and 141 fmole/mg protein respectively using established natural occurring tachykinins to displace the binding. The rank order of potency was identified as substance P > neurokinin A > neurokinin B. These natural occurring tachykinins stimulate inositol phospholipid hydrolysis in slices of rat salivary glands, and the rank order of potency was also substance P > neurokinin A > neurokinin B. When salivation was induced by natural occurring tachykinins and acetylcholine (via i.v. route) in anesthetized rats with doses eliciting equivalent salivating responses, atropine blocks acetylcholine- as well as neurokinin B-, but not substance P- or neurokinin A-induced salivation. Based on the results mentioned above, we make a tentative conclusion that multiple receptor subtypes of neurokinins may exist in rat salivary glands. The neurokinins B receptor subtype is likely located presynaptically in the cholinergic nerve endings, whereas substance P and/or neurokinin A receptor subtypes may exist in the glandular tissue. The salivation induced by these neurokinins is likely through the activation of receptors, which are coupled to phosphatidylinositol turnover pathway.

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.

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.

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.

[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.

Muscarinic M2 receptors coupled to inhibition of adenylate cyclase in rat heart.

The biochemical linking event between the activation of muscarinic receptors and the inhibition of adenylate cyclase was studied with rat heart ventrical membranes. The muscarinic M2 selective antagonists were more potent than the M1 selective antagonists in the displacement of non-selective labeled 3H-QNB binding to the membranes. This was also true for the M2 selective agonists, with respect to the M1 selective agonists, in the reaction medium without Gpp(NH)p. With the same preparation, the muscarinic M2 selective agents were also more potent than the M1 selective agents in the inhibition of adenylate cyclase activity. The order of potencies of these agents in the displacement of 3H-QNB binding correlated well with their order of potencies in inhibiting adenylate cyclase activity. Gpp(NH)p reduced the binding affinities of M2 selective agonists (i.e. carbachol and oxotremorine), while it did not affect the binding affinities of non-selective agonist pilocarpine, M1 selective agonist McN-A-343 and antagonists (i.e. pirenzepine, trihexyphenidyl, AF-DX-116 and methoctramine). These results suggest that in the rat heart, the inhibition of adenylate cyclase by muscarinic agonists is through activation of the M2 subtype receptor and G-protein is likely to be involved in the coupling.

Differential distribution of muscarinic receptor subtypes and their regulation by G-protein in rat brain.

1. [3H]quinuclidinyl benzilate ([3H]QNB) binding in rat cerebral and cerebellar synaptosomes had different Bmax values, but similar Kd values. 2. These bindings could be displaced by classic muscarinic agents: pilocarpine (partial agonist), and atropine (antagonist), which both had similar binding affinities in rat cerebral and cerebellar synaptosomes. 3. The new muscarinic M1 selective agents: McN-A-343 (agonist), pirenzepine and trihexyphenidyl (antagonists) and higher affinities for receptor sites in the cerebrum than in the cerebellum. 4. The muscarinic M2 selective agents: carbachol, oxotremorine (agonists), and AF-DX-116 (antagonist) had higher affinities for receptor sites in the cerebellum than in the cerebrum. 5. GPP(NH)p (40 microM) decreased the binding affinities of carbachol and oxotremorine in the cerebellum, but not in the cerebrum. However, it did not decrease the binding affinities of all the antagonists studied in both brain regions. 6. These results reveal that more muscarinic M1 sites are present in the cerebrum than in the cerebellum, while the opposite is true for M2 sites. Furthermore, the regulatory role of G-protein on these muscarinic receptor subtypes in the brain is different.

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.