Surgical Wound

Postnatal day 49–63 (P49–P63) male and female C57BL/6 mice were used in all experiments in accordance with institutional guidelines. All surgical procedures were conducted under general anesthesia using continuous isoflurane (induction at 5%, maintenance at 1–2.5% vol/vol). Depth of anesthesia was monitored continuously and adjusted when necessary. After induction of anesthesia, the mice were fitted into a stereotaxic frame, with their heads secured by blunt ear bars and their noses placed into an anesthesia and ventilation system (David Kopf Instruments). Mice were administered 0.05 ml buprenorphine (0.1 mg/ml; Buprenex) subcutaneously before surgery. The surgical incision site was then cleaned three times with 10% povidone iodine and 70% ethanol. Skin incisions were made, followed by craniotomies of 2–3 mm in diameter above the left parietal cortex using a small steel burr (Fine Science Tools) powered by a high speed drill (K.1070; Foredom). Saline (0.9%) was applied onto the skull to reduce heating caused by drilling. Unilateral viral injections were performed by using stereotaxic apparatus (David Kopf Instruments) to guide the placement of beveled glass pipettes (World Precision Instruments) into the left hippocampus (2 mm posterior to bregma, 1.5 mm lateral to midline, and 1.6 mm from the pial surface). Either 2 µl AAV2/5 gfaABC₁D Lck-GCaMP3 (1.2 × 10¹³ gc/ml), 1.5 µl AAV2/5 gfaABC₁D GCaMP3 (1.5 × 10¹³ gc/ml), 1.5 µl AAV2/5 gfaABC₁D Lck-GFP (2.41 × 10¹³ gc/ml), or 1.0 µl AAV2/5 gfaABC₁D tdTomato (2.5 × 10¹³ gc/ml) was injected using a syringe pump (Pump11 PicoPlus Elite; Harvard Apparatus). Glass pipettes were left in place for at least 10 min. Surgical wounds were closed with single external 5–0 nylon sutures. After surgery, animals were allowed to recover overnight in cages placed partially on a low voltage heating pad. Buprenorphine was administered two times per day for up to 2 d after surgery. In addition, trimethoprim sulfamethoxazole (40 and 200 mg, respectively, per 500 ml water) was dispensed in the drinking water for 1 wk. Mice were killed 12–20 d after surgery for imaging (typically 13–15 d). We chose this period because generally it takes ∼2 wk to achieve GECI expression in cells by AAV infection and because it has been suggested that long-term expression (>3 wk after AAV injection) can cause toxicity in neurons (Akerboom et al., 2012 (link)).

After research ethics board approval, we conducted a retrospective cohort study of consecutive adults aged ≥18 yr who underwent elective non-cardiac surgery from March 2010 to December 2011 at the University Health Network (Toronto, Ontario, Canada), a quaternary care medical centre offering all adult surgical services except trauma and obstetrics. The cohort included all individuals who underwent elective non-cardiac surgery within 30 days after outpatient assessment at the institutional preoperative assessment clinics. The research ethics board waived the requirement for written informed consent for this study.

The study design was retrospective cohort study. The study proposal was approved by The Human Research Ethics Committee of Thammasat University (Medicine). The patients, who presented with symptomatic GS or complications of GS, then underwent LC since January 2017 to December 2021 in service of Hepato-Pancreato-Biliary and Transplantation unit in surgery department of Thammasat University Hospital, were considered to be enrolled into this study. The electronic medical record was thoroughly reviewed.
The important information including demographic data, clinical presentation, laboratory results, and radiological findings was collected. The operative time, intraoperative findings, perioperative complications, and conversion to open surgery were reviewed from operative notes. The laparoscopic procedure was carried out through three or four small incisions at umbilical and right upper quadrant areas. The operative time was counted from the opening of the first port-site incision to the closure of the last surgical wounds.
Some cases might be excluded because of the following reasons: (1) patients who underwent LC with other indication such as gallbladder polyp, (2) LC was performed in emergency setting for treatment of acute cholecystitis, and (3) there were any other procedures performed in the same setting of LC such as intraoperative ERCP. By the perioperative information, the patients were categorized into three groups by difficulty grading as given in Table 2.
The univariate analysis was performed using chi-square test for categorical data and Student's t-test for continuous data to define the significant factors affecting on very difficult LC and converted cases. Then multivariate analysis was carried out for both outcomes. Thereafter, the preoperative predictive scores of each patient were calculated using the original Randhawa scoring systems and also the modification of Tongyoo et al. The comparison between scores from both models was performed by many methods such as paired t-test, correlation coefficient, and area under receiver operating characteristic (ROC) curve. All of statistical analyses were performed by IBM SPSS^® Statistics version 20 and their results were determined to be significant at P < .05.