Thiopental Sodium

Embryo collection was performed in a surgical room located on the farm. The donors were subjected to a midventral laparotomy on Days 5 and 6 of the estrous cycle (Day 0: onset of estrus) to obtain morulae and unhatched blastocysts, respectively. The donors were sedated by the administration of azaperone (2 mg/kg body weight, intramuscular). General anesthesia was induced using sodium thiopental (7 mg/kg body weight, intravenous) and maintained with isoflurane (3.5–5%). After exposure of the genital tract, the corpora lutea on the ovaries were counted. Embryos were collected by flushing the tip of each uterine horn with 30 mL of a chemically defined medium consisting of Tyrode's lactate (TL)-HEPES-polyvinyl alcohol (PVA) [17] (link) with some modifications. This medium (TL-PVA) was composed of 124.3 mM NaCl, 3.2 mM KCl, 2 mM NaHCO₃, 0.34 mM KH₂PO₄, 10 mM Na-lactate, 0.5 mM MgCl₂·6H₂O, 2 mM CaCl₂·2H₂O, 10 mM HEPES, 0.2 mM Na-pyruvate, 12 mM sorbitol, 0.1% (w/v) PVA, 75 µg/ml potassium penicillin G and 50 µg/mL streptomycin sulfate. The collected embryos were evaluated to verify their developmental stage and quality grade. One-cell eggs and poorly developed embryos were classified as oocytes and degenerate embryos, respectively. The remaining embryos that exhibited appropriate morphology according to the criteria determined by the International Embryo Transfer Society [18] were considered viable. Only compacted morulae and unhatched blastocysts graded as excellent or good based on morphological appearance were used in the experiments according to the specific experimental design.
The ovulatory response of the donors was determined by counting the number of corpora lutea in both ovaries. To evaluate the effectiveness of the superovulation treatment, the numbers of viable embryos and oocytes and degenerate embryos were counted in each donor. The recovery rate was defined as the ratio of the number of embryos and oocytes and degenerate embryos recovered to the number of corpora lutea present. The fertilization rate was defined as the ratio of the number of viable embryos to the total number of embryos and oocytes and degenerate embryos collected.

The collection of embryos was performed in a specifically designed surgical room located on the farm. The donors were subjected to a mid-ventral laparotomy on Day 6 of the estrous cycle (Day 0: onset of estrus). The donors were sedated with azaperone (2 mg/kg body weight, intramuscular). General anesthesia was induced using sodium thiopental (7 mg/kg body weight, intravenous) and was maintained with isoflurane (3.5-5%). After exposure of the genital tract, the corpora lutea on the ovaries were counted. The embryos were collected by flushing the tip of each uterine horn with 30 mL of Tyrode’s lactate (TL)-HEPES-polyvinyl alcohol (PVA)15 (link) with some modifications4 . The recovered embryos were evaluated under a stereomicroscope at a magnification of 60× to grade the developmental stage and quality. One-cell eggs and poorly developed embryos were classified as unfertilized oocytes and degenerated embryos, respectively. The remaining embryos with the appropriate morphology according to the criteria determined by the International Embryo Transfer Society16 were considered viable. Vitrification was only performed on compacted morulae and unhatched blastocysts with morphology graded as excellent or good.
The ovulatory response of the donors was determined by counting the number of corpora lutea on both ovaries. Recovery rate was defined as the ratio of the number of embryos and oocytes and degenerated embryos recovered to the number of corpora lutea present. Fertilization rate was defined as the ratio of the number of viable embryos at collection to the total number of embryos and oocytes and degenerated embryos collected.

At 126 ± 2 days gestation (term ∼ 148 days), Border-Leicester ewes were anaesthetised with an intravenous bolus of 5% sodium thiopentone (Pentothal; 1g in 20 ml) and, following intubation, maintained with inhalation of 1.5–3% halothane in air. The fetal head and neck were exposed via caesarean section and an ultrasonic flow probe (3 mm: Transonic Systems, Ithaca, NY, USA) was placed around a carotid artery. Heparinised saline-filled polyvinyl catheters were inserted into the other carotid artery and into a jugular vein. The fetal trachea was intubated with a 4.0 mm cuffed endotracheal tube and lung liquid was drained passively for ∼ 10 seconds or until liquid ceased exiting the airways. A transcutaneous arterial oxygen saturation (SpO₂) probe (Masimo, Radical 4, CA, USA) was placed around the right forelimb and the output recorded continuously. A Near Infrared Spectroscopy optode (Casmed Foresight, CAS Medical Systems Inc, Branford, CT, USA) was placed over the left frontal cortex and used to continuously measure cerebral tissue oxygen saturation (SctO₂). After completion of instrumentation, the ewe was rotated onto its side and the fetus was completely exteriorised from the uterus, still attached to the umbilical cord, dried, and placed on a delivery table immediately next to the ewe. Physiological parameters were allowed to stabilise prior to the birth procedures being initiated (see below)—the delay between exteriorisation and intervention was a mean (SD) of 181 ± 31 seconds for all lambs.
Each fetus was randomised to either umbilical cord clamping prior to initiation of ventilation (Clamp 1^st; n = 10) or the initiation of ventilation prior to umbilical cord clamping (Vent 1^st; n = 7) groups. In Clamp 1^st lambs, the umbilical cord was immediately clamped and cut, the lamb transferred to an infant warmer (CosyCot, Fisher and Paykel, Auckland, New Zealand) and ventilation commenced as soon as possible. In Vent 1^st lambs, ventilation commenced while the umbilical cord remained patent. Umbilical cord clamping was delayed until after PBF had increased, indicating cardiopulmonary transition, whereupon the cord was clamped and cut. In both groups ventilation was initiated using positive pressure ventilation in volume guarantee mode with a tidal volume of 7 mL/kg and a positive end-expiratory pressure of 5 cmH₂O using warmed and humidified inspired gases, with an initial fraction of inspired oxygen (FiO₂) of 21% (Babylog 8000+, Dräger, Lübeck, Germany). Peak inspiratory pressure was limited to 40 cmH₂O to avoid pneumothoraces. FiO₂ was adjusted to target SpO₂ at 70–90% by 3 min, 80–90% by 5 min and 90–96% by 10 min.
Regular blood gas analysis (ABL30, Radiometer, Copenhagen, Denmark) and real-time SpO₂ measurements were used to monitor the lambs. All lambs received sedation (Alfaxane i.v. 5–15 mg/kg/h; Jurox, East Tamaki, Auckland, New Zealand) in 5% dextrose via the jugular vein catheter to minimize spontaneous breathing during the experiment. The ewes were humanely euthanized using sodium pentobarbitone (100 mg/kg i.v) after caesarean section and the lambs were euthanized after completion of the ventilation study, at 30 min—2 h later, depending on the study.

As shown in Fig. 2, rats were randomly divided into six groups with eight rats in each group, including the control group, model group, baicalin magnesium groups (50 mg/kg and 150 mg/kg), baicalin group (146.4 mg/kg) and MgSO₄ group (19.7 mg/kg). Baicalin magnesium and MgSO₄ were dissolved in sterilized water for injection. Baicalin was added to sterilized water for injection and then adjusted to pH 7.4 with 10% NaOH. The concentrations of baicalin magnesium, baicalin and MgSO₄ were 97 mg/ml, 188 mg/ml and 40 mg/ml, respectively. The baicalin group was administered a dose that was equimolar to the baicalin parent nucleus of baicalin magnesium (150 mg/kg), and the MgSO₄ group was administered equimolarly to the magnesium ion of baicalin magnesium (150 mg/kg). The tail vein of the control and model groups were injected with the same amount of saline as the administered group. After 1 week of normal feeding adaptation, rats (with the exception of the control group) were fed with HFD for 8 weeks to develop NASH, followed by 2 weeks of continuous HFD feeding. The rats were anesthetized by intraperitoneal injection of 1% thiopental sodium (50 mg/kg) [19 (link)], and the livers were removed to calculate the liver index and the ratio of liver mass to body mass. Blood was also collected for subsequent experiments.Fig. 2

Timeline demonstrating the induction of NASH and treatment. (BA-Mg: baicalin magnesium; BA: baicalin)

The FBG measurements were made using a glucometer (one-touch Verio flex). Rats with FBG levels above 200 mg/dl with signs of polyuria and polydipsia were confirmed to be diabetic and were involved in the study. The following treatment was given in oral dosing using a gavage tube to all groups for 28 days. Rats in Groups I and diabetic rats in Group II were administered orally with distilled water. Group III diabetic rats were treated with Glibenclamide standard, 0.6 mg/kg b.w., and bitter honey was administered to diabetic rats in Groups IV and V, 200 mg/kg b.w. and 400 mg/kg b.w., respectively. The Glibenclamide standard solution was prepared in 1% Carboxymethyll cellulose, while bitter honey samples were prepared by diluting in distilled water. After the experimental period (24 hours after the last dose), the animals were euthanized using an excess dose of anesthesia (thiopentone sodium, 75 mg/kg ip) [26 (link)].