[Introduction and development of the Trans Woman Voice Questionnaire authorized simplified Chinese translation].

Objective: The purpose of this article is to translate and adapt the Trans Woman Voice Questionnaire (TWVQ) into the simplified Chinese version (TWVQ-SC), and to evaluate its reliability and validity. Methods: Authorized by the author of the TWVQ,the TWVQ-SC was developed through translation, back translation,and cross-cultural adaptation.The TWVQ-SC contained 30 items capturing personal perception of vocal function, psychosocial impact of voice, and degree of limitation in social participation. Subjects included 279 trans women in the experimental group, 128 cis women in the control group, and 89 trans women in the retest group. The Cronbach α and the item total correlation coefficient (ITC) were calculated to examine the internal consistency. The intraclass correlation coefficient (ICC) was chosen to examine the test-retest reliability. Regarding validity, the expert judgment method was utilized to examine the content validity. Factor analysis and Spearman's rank correlation coefficient were used to examine the construct validity, and the discriminant validity was examined by the rank sum test of the total scores of the cisgender and transgender subjects. Results: The Cronbach's α of TWVQ-SC is 0.97 and the ITC of 30 items range from 0.40 to 0.86. The ICC is 0.84. The four principal components' cumulative contribution is 65.12%. The Spearman rank correlation coefficient to VHI-10 is 0.85 (P<0.01). The total score of the TWVQ scale in the transgender female group is significantly higher than that in the cisgender female group (U=1 580,P<0.01). Conclusion: TWVQ-SC demonstrates good reliability and validity and therefore can be used clinically as a self-assessment tool for transgender women to evaluate their own voice.

Gamma-hydroxybutyric acid-induced absence seizures in GluR2 null mutant mice.

In this electrophysiological study, we examined the susceptibility of GluR2 mutant null mice to absence seizures in comparison with wild-type controls. The prodrug of (GHB), gamma-butyrolactone (GBL) was given systemically to induce the absence seizures. We also tested the severity and duration of the seizure activity in this model. The results showed that the latency from GBL administration to onset of seizure was significantly prolonged in GluR2(-/-) mice when compared to GluR2(+/+) mice. The duration of spike-and-wave discharges (SWD) was also significantly decreased in the GluR2(-/-) mice. Ninety minutes following GBL administration, wild-type animals continued to exhibit intermittent SWD bursts while GluR2(-/-) mice had returned to baseline. These data suggest that the GluR2 subunit may be involved in the initiation and maintenance of absence seizures induced by GBL.

Alteration of GLUR2 expression in the rat brain following absence seizures induced by gamma-hydroxybutyric acid.

We explored the involvement of the glutamate receptor subunit B (GluR2) in the mechanism of absence seizures induced by gamma-hydroxybutyric acid (GHB). The expression and distribution of GluR2 protein in rat brain were examined during and after GHB-induced absence seizures. The data indicate that GluR2 protein expression significantly decreases following the onset of absence seizures. The suppression of GluR2 expression was prolonged and it outlasted the duration of the continuous absence seizure activity. The alteration of GluR2 protein levels was accompanied by a re-distribution of GluR2 expression from laminae V to IV in cerebral cortex. We also analyzed the duration and latency of absence seizures induced by GHB 72 h following an initial GHB-induced absence seizure, a time when suppression of GluR2 protein was maximal. The second absence seizure was significantly more prolonged than the first. These data may indicate that the putative down-regulation of GluR2 following GHB-induced absence seizure could have contributed to the potentiation of subsequent seizures in animals. A related hypothesis posed by the data is that down-regulation of GluR2 is involved in the mechanisms of the maintenance of recurrent absence seizure activity once it is initiated and therefore, may contribute to the chronicity of seizures in absence epilepsy.

[Relationship between exposure time and cryopreservation of articular cartilage].

OBJECTIVE: To search an optimal method for improving viability of cryopreserved articular cartilage. METHODS: Articular cartilage which was sampled from the rabbits were randomly divided into 5 groups. Fresh cartilage was group I, other groups were frozen. Before frozen, other cartilage was exposured in 10% DMSO at 4 degrees C for 30 minutes(group II), 1 hour(group III), 2 hours (group IV), 4 hours(group V), then were stored in liquid nitrogen for 1 week. Viabilities of the chondrocytes were detected by Typan-blue staining, electron transmission microscope, and determination of incorporation 3H-TdR after the temperature returned to normal. RESULTS: 1. The cells were injuried at different extent after the cartilage was frozen. In group I, survival rate of cells was 96% and incorporation of 3H-TdR was (4,953.13 +/- 583.27)%, statistic difference was significant between group I and other groups(P < 0.01). The microstructure of group I was normal while other groups all had damage of the organella, 2. Structures and functions of cells in group IV were best among frozen groups. Organella were less damaged than group II, III, V, survival rate of cells was 56% and incorporation of 3H-TdR was (1,139.88 +/- 146.39)%, statistic difference was significant between group IV and group II, III, V(P < 0.01). CONCLUSION: If cartilage are exposured in 10% DMSO at 4 degrees C for 2 hours before frozen, optimal cryopreservation can be achieved.

Regulation of gamma-aminobutyric acid (GABA) release in cerebral cortex in the gamma-hydroxybutyric acid (GHB) model of absence seizures in rat.

Gamma-hydroxybutyric acid (GHB) has the ability to induce absence seizures. The precise way in which GHB causes seizures remains unclear, but GABA(B)- and/or GHB-mediated presynaptic mechanisms within thalamocortical circuitry may play a role. In the present study, we determined the basal and K+-evoked release of GABA and glutamate in the superficial laminae of frontal cortex during GHB-induced absence seizures. Our data indicate that both the basal and K+-evoked release of GABA were significantly decreased in laminae I-III of frontal cortex at the onset of GHB-induced absence seizures. The appearance and disappearance of the observed changes in basal and K+-evoked extracellular levels of GABA correlated with the onset and offset of absence seizures. In contrast, neither the basal nor the K+-evoked release of glutamate was altered in superficial laminae of cerebral cortex at any time during the absence seizures. Intracortical perfusion of the GABA(B) receptor antagonists, CGP 35348 and phaclofen as well as the GHB receptor antagonist, NCS 382 attenuated GHB-mediated changes in the basal and K+-evoked release of GABA. These data suggest that GHB induces a selective decrease in the basal and depolarization-induced release of GABA in cerebral cortex, and further, that this action of GHB may play a role in the mechanism by which GHB induces absence seizures.

Neuronal stress and injury in C57/BL mice after systemic kainic acid administration.

Kainate-induced seizures are widely studied as a model of human temporal lobe epilepsy due to behavioral and pathological similarities. While kainate-induced neuronal injury is well characterized in rats, relatively little data is available on the use of kainate and its consequences in mice. The growing availability of genetically altered mice has focused attention on the need for well characterized mouse seizure models in which the effects of specific genetic manipulations can be examined. We therefore examined the kainate dose-response relationship and the time-course of specific histopathological changes in C57/BL mice, a commonly used founder strain for transgenic technology. Seizures were induced in male C57/BL mice (kainate 10-40 mg/kg i.p.) and animals were sacrificed at various time-points after injection. Seizures were graded using a behavioral scale developed in our laboratory. Neuronal injury was assayed by examining DNA fragmentation using in situ nick translation histochemistry. In parallel experiments, we examined the expression an inducible member of the heat shock protein family, HSP-72, another putative marker of neuronal injury, using a monoclonal antibody. Seizure severity paralleled kainate dosage. At higher doses DNA fragmentation is seen mainly in hippocampus in area CA3, and variably in CA1, thalamus and amygdala within 24 h, is maximal within 72 h, and is largely gone by 7 days after administration of kainate. HSP-72 expression is also highly selective, occurring in limbic structures, and it evolves over a characteristic time-course. HSP-72 is expressed mainly in structures that also manifest DNA fragmentation. Using double-labeling techniques, however, we find essentially no overlap between neurons expressing HSP-72 and DNA fragmentation. These findings indicate that DNA fragmentation and HSP-72 expression are complementary markers of seizure-induced stress and injury, and support the notion that HSP-72 expression is neuroprotective following kainate-induced seizures.

Glutamate receptor antagonists reduce spontaneous epileptiform activity in cortical wedges prepared from DBA/2 mice.

This study investigated the involvement of glutamatergic neurotransmission in the epileptiform activity demonstrated in cortical weges prepared from genetically audiogenic seizure-prone DBA/2 mice. Omission of Mg2+ from the perfusing medium initiated spontaneous epileptiform events with accompanying afterpotentials on the repolarizing phase. These spontaneous depolarizations also occurred in some 30% of the slices in the presence of Mg2+ (2 mM). The N-methyl-D-aspartate (NMDA) receptor antagonist 3-(2-carboxypiperazin-4-yl)-propenyl-1-phosphonic acid (CPP) and the non-competitive NMDA receptor channel blocker ketamine produced a significant reduction of these spontaneous depolarizations. 7-Chlorokynurenic acid (7-CKA), an antagonist at the strychnine-insensitive site on the NMDA receptor, also exerted an inhibitory effect. In addition the AMPA/kainate receptor antagonist 6,7-dinitroquinoxaline-2,3-dione (DNQX) suppressed the spontaneous events. These observations provide evidence that glutamatergic neurotransmission contributes to the epileptiform activity in this cortical preparation.

Tigabine hydrochloride, an inhibitor of gamma-aminobutyric acid (GABA) uptake, induces cortical depolarizations in vitro.

The effect of the gamma-aminobutyric acid uptake inhibitor tiagabine hydrochloride was studied on electrical responses in cortical wedges prepared from 20-30 day-old, audiogenic seizure-prone DBA/2 mice. Perfusion of tiagabine (50 microM) for 15 min, evoked large, slow depolarizations with a frequency of 6-8/h which persisted for 4-5 h. The GABA(A) receptor antagonists, bicuculline (10 microM) and picrotoxin (100 microM), inhibited established depolarizations. These depolarizations were also calcium-dependent and blocked by tetrodotoxin. The non-opioid antitussive, dextromethorphan, which has been shown to inhibit glutamate release, irreversibly blocked the depolarizations. Conversely, 4-aminopyridine (50 microM), a potassium channel antagonist, markedly potentiated the responses. The NMDA receptor antagonist, 3-((R)-2-carboxypiperazin-4-yl)-propyl-1-phosphonic acid, had no effect on the depolarizations at concentrations up to 100 microM but the AMPA/kainate receptor antagonist, 6,7-dinitroquinoxaline-2.3-dione at high concentrations (100 and 200 microM), reversibly decreased the frequency without affecting the amplitude. It is concluded that the tiagabine-induced depolarizations in this in vitro preparation were initiated through GABA(A) receptors leading, possibly, to a release of excitatory amino acids.

Crystal structure of an acidic neurotoxin from scorpion Buthus martensii Karsch at 1.85 A resolution.

The crystal structure of an acidic scorpion neurotoxin, BmK M8, purified from Chinese scorpion Buthus martensii Karsch (BmK), has been determined by the molecular replacement method. It is the first structure of an acidic alpha-scorpion neurotoxin reported so far. The crystals adopt a symmetry of space group P2(1) and contain one molecule per asymmetric unit. The structure has been refined to an R factor of 18.1% using reflection data in the range of 8 to 1.85 A resolution, with standard deviations from ideal geometry of 0.017 A and 2.43 degrees for bond length and angle, respectively. The 12 residues at the C terminus with unknown sequence were determined by crystallographic refinement. The refined model shows that the structural core, consisting of a motif beta alpha beta beta, is similar to that of toxin II from Androctonus australis Hector (AaH II) or Variant 3 from Centruroides sculpturatus Ewing (CsE V3). The three conformationally variable loops protruding from this structural core are different from that of AaH II, and especially from that of CsE V3. Compared with the most potent and basic alpha-toxin AaH II, the BmK M8 is a relatively inactive toxin (1100 times less active than AaH II) with an unusually low isoelectric point (pI 5.3). Sequence alignment of the two toxins shows a difference of 26 residues (40.6%). Among them four basic or neutral residues in AaH II, namely Val10, Lys28, Val55 and Gly59, are changed to acidic glutamate in BmK M8. The residues Glu10, Glu28 and Glu55 of BmK M8 are located on a surface (Face B), opposite the "conserved hydrophobic surface" (Face A). The latter is a functionally important area proposed by Fontecilla-Camps et al. Our observations suggest that in addition to Face A, Face B may also be involved in the biological activity of scorpion toxins. The structure of BmK M8 shows an evident conformational change of the alpha-amino group at the N terminus and a deorganization of Arg2 caused by the mutation D53A. These structural changes may also be responsible for the weak toxicity of BmK M8. In association with the information from chemical modifications, a multisite binding mode for toxin-receptor interaction and three "toxic regions" in relevance to the binding process, including Face A, Face B and Site C, are proposed. Face A, mainly consisting of Tyr5, 35, 47, the alpha-amino group, Arg2 and Asp3, may be more essential for the binding. Face B, mainly comprising conserved residues Tyr14, 21, Lys28 and Val55, may contribute to the high efficacy of the binding process and substitutions by acidic residues in this area could strongly weaken the toxic activity. Site C, formed by Lys58 and Arg62 at the C terminus and Arg41 and Tyr42 from loop 38-44, may be involved in binding site specificity.

The effect of the desglycinyl metabolite of remacemide on cortical wedges prepared from DBA/2 mice.

Remacemide hydrochloride is currently undergoing clinical trials for use as an anticonvulsant agent in the treatment of epilepsy. It is considered that the desglycinyl metabolite (FPL 12495AA) of the parent compound accounts for the majority of the anticonvulsant activity. In this study we have investigated the effects of FPL 12495AA on electrical activity in the cortical wedges prepared from audiogenic seizure-prone DBA/2 mice. FPL 12495AA at varying concentrations (50-200 microM) significantly reduced both the spontaneous depolarizations (IC50 102 microM) and the associated afterpotentials (IC50 50 microM) which are characteristic in this preparation under magnesium-free conditions. The compound also concentration-dependently reduced N-methyl-D-aspartate (NMDA)-induced depolarizations of the tissue (IC50 43 microM) and the antagonism by FPL 12494AA was not overcome by increasing NMDA concentrations. FPL 12495AA had no effect on (S)-alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid (AMPA)-induced depolarizations. The results suggest that FPL 12495AA has a specific antagonistic effect on the NMDA receptor complex possibly through non-competitive inhibition at the phenycyclidine site in the ion channel. Such an action could contribute to its anticonvulsant properties.

Purification and characterization of multiple components of human lymphoblastoid interferon-alpha.

Twenty-two components of human interferon-alpha (IFN-alpha) derived from Sendai virus-induced Namalwa cells were purified by sequential immunoadsorbent affinity chromatography using four monoclonal antibody affinity columns followed by ultrafiltration and reversed-phase high-performance liquid chromatography. The specific activity ranged from 0.2 to 2.6 x 10(8) IU/mg protein on Madin-Darby bovine kidney cells, 0.3 to 4.6 x 10(8) IU/mg protein on human WISH cells, and 10(4) to 7 x 10(5) units/mg protein on mouse L929 cells. The apparent molecular weights of the components ranged from 17,500 to 23,300 using nonreducing sodium dodecyl polyacrylamide-gel electrophoresis and 17,500 to 27,600 using reducing sodium dodecyl polyacrylamide-gel electrophoresis. The amino-terminal amino acid sequences were similar among the components as well as to those reported for the cloned human IFN-alpha genes (Pestka, S. (1986) Methods Enzymol. 119, 3-14). However, four components, f, i, l, and m, have amino-terminal amino acid sequences which appear to be unique when compared to those predicted from the cDNA clones. One component, pre-a, has a potential N-linked glycosylation site on the Asn of residues 2 through 4, Asn-Leu-Ser.

Modulation of epidermal growth factor receptors by human alpha interferon.

Treatment of Madin-Darby bovine kidney (MDBK) cells with human interferon (IFN)-alpha 2 at 37 degrees C results in a dose-dependent inhibition of cell growth and a reduction of the subsequent binding of 125I-labeled epidermal growth factor (EGF) at 4 degrees C. Human IFN-beta and -gamma, which exhibit little antiviral and antiproliferative activities on MDBK cells, have little effect on cell growth or the binding of 125I-labeled EGF to these cells. The binding of EGF is decreased after exposure to IFN-alpha for greater than 8 hr. Scatchard analyses of the EGF binding data indicate that a 20-hr exposure period results in a decrease in the apparent number of cell-surface EGF receptors and a reduction in the affinity of EGF for its receptor. The rate of internalization of EGF by MDBK cells does not appear to be affected by IFN treatment.

Microlymphaticovenous anastomosis for lymphedema of external genitalia in females.

Lymphedema of the female external genitalia is seldom reported and can be psychologically distressing to the patient. On the basis of microlymphaticovenous operations for lymphedema of the extremities, a new microsurgical operation has been developed for treating lymphedema of the female external genitalia and applied clinically in six patients with good results. Edema of the female external genitalia and of the lower part of the abdominal wall has been reduced postoperatively, the chylorrhea has disappeared and the blisters on the vulva have vanished. The operative techniques and the anatomic basis of the operation are described, the six patient histories are illustrated and the correlation between number of anastomoses and quality of results are discussed herein.

Microlymphaticovenous anastomosis in the treatment of lower limb obstructive lymphedema: analysis of 91 cases.

Since 1980, 110 cases of lymphedema have been treated by microlymphaticovenous anastomosis. Of these 110 patients, 91 with obstructive lymphedema of lower limbs were reviewed. The immediate and long-term results have been very satisfactory. Excellent and good results were obtained in 79.1 percent. An average reduction in circumference of the affected limb of 6.4 cm and an average reduction of excess volume of 59.2 +/- 29.5 percent (representing 703 +/- 850 ml) were obtained. Subjective symptoms and objective signs were improved. Four patients (4.4 percent) showed poor results owing to severe fibrosis of neighboring tissue; no lymphatics could be located for anastomosis. As the authors gained experience with the operation over the last 3 years, they modified the operative procedure, the anastomotic technique, and the selection of collective lymphatics. The data obtained suggest that the quality of results is proportional to the number of anastomoses. In order to obtain the best results, the criteria for selection of patients and avoidance of postoperative relapse are discussed. Finally, a test for determination of the indications for microlymphatic surgery is described.

Influence of vessel clips on vascular patency in microarterial grafts less than 0.5 mm in external diameter.

Autogenous interpositional microarterial grafting of 140 saphenous branches of albino rat femoral arteries with an external diameter of 0.3 to 0.5 mm (mean 0.45 +/- 0.06 mm) resulted in an immediate patency rate of 100%. The patency rate observed at 72 hours was 64.3%. The late patency rate was related to blade tip pressure of the double approximator clip used in the microanastomosis. The patency rate with a clip with a blade tip pressure of 22 gm was 52.7% and that with a clip with a blade tip pressure of 2 gm was 77.3%. When arterial occlusion lasted less than 30 minutes, the late patency rate was 76.9% to 78.3%. If arterial occlusion lasted for as long as 1 hour, the late patency rate of vessels occluded by a clip with a blade tip pressure of 22 gm dropped to 27.8%. These results demonstrate that an interpositional microarterial graft as small as 0.5 mm in external diameter is clinically feasible.

Microlymphaticovenous anastomosis for lymphedema of the breast.

On the basis of microlymphaticovenous anastomosis for treating lymphedema of the extremities, the authors developed a microlymphaticovenous procedure to treat congenital lymphedema of the breast and applied it clinically in two patients. The immediate results have been fairly good. The breast and its nipple had been reduced in size and to a nearly normal level. The results are maintained after a half year of follow-up. The fundamentals and techniques of the operation are described and the two case histories are reported. The cases presented here appear to be the first successful clinical attempts to treat lymphedema of the breast by microlymphaticovenous anastomosis. The clinical character of congenital lymphedema, the indication of the microlymphaticovenous anastomosis, and the factors required for success of microlymphatic surgery are discussed.

Microlymphaticovenous anastomosis for treating scrotal elephantiasis.

Scrotal elephantiasis can be physically disabling and psychologically distressing to the victim. Ablative procedure has been used in its treatment and has achieved limited success. The authors developed a microlymphaticovenous procedure to treat elephantiasis of the scrotum and applied it clinically in three patients. The immediate and long-term (13-24 months) results have been very satisfactory. The scrotum size was dramatically reduced to a nearly normal level, and subjective symptoms and objective signs were improved. The operative techniques are described, the three case histories are illustrated, and the advantages of microlymphaticovenous anastomosis, the selection of patients, and the factors required for success of the surgery are discussed.

Microvascular free transfer of iliac bone based on the deep superior branches of the superior gluteal vessels.

The findings from dissection, injection studies, corrosion specimens, and angiograms in 300 specimens of the iliogluteal region taken from 150 cadavers are reported. These findings suggest that the deep superior branches of the superior gluteal vessels may serve a nutritive pedicle in microvascular free transfer of iliac bone. Therefore, the operative technique has been developed and applied clinically with good results. The selection of different vessels as a nutritive pedicle for the free transfer of iliac bone is discussed.

Microvascular free transfer of iliac bone based on the deep circumflex iliac vessels.

The findings from dissections, injection studies, and angiograms of 180 specimens of the anterior portion of left and right iliac crests taken from 90 cadavers are reported. These findings suggested that the deep circumflex iliac vessels were suitable for use as a nutritive pedicle in microvascular free transfer of iliac bone. Therefore, the technique of microvascular free transfer of iliac bone based on these vessels was developed and applied clinically in 2 cases with good results. The advantages and disadvantages of using the superficial circumflex iliac vessels or the deep circumflex iliac vessels as the vascular pedicle for iliac grafts also are discussed.