A novel three-step DNA extraction method for mixed bloodstains.

Mixed DNA samples from at least two contributors can be present at a crime scene, which could be the most crucial piece of genetic evidence. The mixed stains in sexual assault cases are typically separated using differential lysis procedures (a two-step method). Blood mixed stains, however, are usually difficult to separate. In this work, we propose that a mixed stain comprises three layers, that is, (1) the upper layer which is primarily made up of cells from one contributor; (2) the middle layer which is a similar mixture from two contributors; and (3) the lower layer which primarily comprises cells from the other contributor. Based on this concept, a novel three-step DNA extraction method was proposed to solve the challenge involving bloodstains from two contributors. In the experiment, we extracted three layers DNA from mixed bloodstains using three steps. As a result, single-source DNA and approximate single-source DNA were detected from steps 1 and 3, respectively. This study demonstrates that the DNA from some mixed blood stains could be effectively separated following an appropriate extraction strategy, providing valuable insights, and serving as a reference for future examination of blood mixtures.

Three-step operation for esophago-left bronchial fistula with respiratory failure after esophagectomy: a case report with literature review.

BACKGROUND: The development of esophago-bronchial fistula after esophagectomy and reconstruction using a posterior mediastinal gastric tube remains a rare complication associated with a high rate of mortality. CASE PRESENTATION: A 63-year-old man with esophageal cancer underwent a thoracoscopic esophagectomy with two-field lymph node dissection and reconstruction via a gastric tube through the posterior mediastinal route. Postoperatively, the patient developed extensive pyothorax in the right lung due to port site bleeding and hematoma infection. Four months after surgery, he developed an esophago-left bronchial fistula due to ischemia of the cervical esophagus and severe reflux esophagitis at the site of the anastomosis. Because of respiratory failure due to the esophago-bronchial fistula and the history of extensive right pyothorax, right thoracotomy and left one-lung ventilation were thought to be impossible, so we decided to perform the surgery in three-step systematically. First, we inserted a decompression catheter and feeding tube into the gastric tube as a gastrostomy and expected neovascularization to develop from the wall of the gastric tube through the anastomosis after this procedure. Second, 14 months after esophagectomy, we constructed an esophagostomy after confirming blood flow in the distal side of the cervical esophagus via gastric tube using intraoperative indocyanine green-guided blood flow evaluation. In the final step, we closed the esophagostomy and performed a cervical esophago-jejunal anastomosis to restore esophageal continuity using a pedicle jejunum in a Roux-en-Y anastomosis via a subcutaneous route. CONCLUSION: This three-step operation can be an effective procedure for patients with esophago-left bronchial fistula after esophagectomy, especially those with respiratory failure and difficulty in undergoing right thoracotomy with left one-lung ventilation.

Application of ISO/IEC Guide 51 to COVID-19 infection control for the occupational safety.

COVID-19 is around the world. We attempt to apply three-step method in ISO/IEC Guide 51: 2014 to COVID-19 infection control in the workplace. The results show that the COVID-19 infection control measures include the eradication of the virus, the destruction of infectivity, the detoxification and weakening and the elimination of opportunities for infection as "Inherently Safe Design Measures", the avoidance of contact as "Safeguarding and Complementary Protective Measures" and the reduction of contact and the avoidance of seriousness as "Information for Use". Among these specific measures, the New Normal, especially in the manufacturing industries, would be "telecommuting" and "unmanned workplaces", which are part of the elimination of opportunities for infection, and "changes in flow lines" and "changes in airflow", which are part of the avoidance of contact. Where "telecommuting" and "unmanned workplaces" are feasible, they should be implemented as much as possible, and where they are not, attempts should be made to minimize human-to-human contact by "changes in flow lines". In addition, in the area of "changes in airflow", there are high expectations for future research on how to establish a ventilation design for COVID-19, in which but also the source would be workers themselves, not only combustible gases and toxic gases.

The methods and techniques of identifying renal pedicle vessels during retroperitoneal laparoscopic radical and partial nephrectomy.

BACKGROUND: Retroperitoneal laparoscopic radical and partial nephrectomy (RLRN and RLPN) have become the preferred modes of management for renal malignancy. One of the most critical steps in the RLRN and RLPN process is to seek and control the renal pedicle. The current study focuses on introducing methods and techniques that can help quickly and accurately identify the renal pedicle vessels during RLRN and RLPN. METHODS: RLRNs and RLPNs were performed for 292 cases in our hospital from November 2014 to January 2017. Different measures were adopted to seek and manage bilateral renal pedicle vessels. All operation procedures were performed by the following three steps: dissection, opening, and clamping. For the left lateral, after the perirenal fat in the dorsal and lateral side was fully dissected, the kidney was pushed toward the ventral side. The renal artery was visible when opening the dense bulging connective tissue, which was located in the middle of the dorsal interior of the kidney. Then, the renal artery was clamped with a Hem-o-lok or the Bulldog clamp. For the right kidney pedicles, the inferior vena cava was first identified and then dissipated upward. When the inferior vena cava was not visible, it was often the location of the right renal artery. The treatment for the artery was the same as for the left renal artery. Relevant clinical characteristics of patients, such as operative time, intraoperative blood loss, and duration of postoperative drainage, were analyzed retrospectively. The three-step method of identifying renal pedicle vessels during retroperitoneal laparoscopic radical and partial nephrectomy was evaluated. RESULTS: All operations were successfully accomplished with satisfying results, during which the artery could be controlled quickly, and no cases were converted to open surgery due to severe bleeding of renal pedicle vessels. There were no complications involving renal vessels during the entire study. The mean operative times were (81.9 ± 19.71) min and (88.2 ± 21.28) min for RLRN and RLPN, with an average intraoperative blood loss of (91.7 ± 47.10) ml and (62.4 ± 47.45) ml, respectively. The warm ischemia time for RLPN was (19.3 ± 5.6) min. The postoperative drainage-tube was removed within (4.5 ± 1.29) d (RLRN) and (4.6 ± 1.98) d (RLPN); the mean postoperative hospital stay times were (7.0 ± 2.4) d and (5.9 ± 1.98) d, respectively. CONCLUSION: The three-step method of identifying renal pedicle vessels during RLRN and RLPN is direct and feasible, and it may help simplify the operating procedure and improve the safety of the surgery. It may be of great practical application value in the clinical field.

Establishing A Method to Rapidly Remove Glycerol in Frozen Red Blood Cells / 现代检验医学杂志

Objective To establish a method to rapidly remove glycerol in frozen red blood cells(RBC).Methods The glycerol in frozen RBC was removed by using three-step method as following:the frozen RBC were diluted by adding salt solution with high concentration.The salt solution was removed through centrifugation.The RBC were resuspended in normal saline of corresponding volume.Three-step method could acbieve the quick removal of glycerol by controlling tbe concentration salt water and action time.The indexes of hemoglobin,free hemoglobin,glycerol residue,residual white blood cell(WBC) and hematocrit of frozen-deglycerolized RBC were compared between conventional and improved methods.Results The traditional method to remove glycerol needed 1h approximately,while the three step method only needed 15 mins approximately,greatly shortening the time and improving the efficiency.The related tests for removing glycerol RBC,except hemoglobin content was slightly lower than the "The quality requirements for whole blood and blood components (2012 edition)",the rest of them all meeted the requirements.Conclusion The three-step method has the advantages of short time and high cfficiency.However,it still needs to be further improved and optimized to make the quality of product meet the national standard.

Clinical Observations on Knife Needle Microinvasive Operation for Cervical Spondylotic Arteriopathy / 上海针灸杂志

Objective To investigate the clinical efficacy of knife needle microinvasive operation for cervical spondylotic arteriopathy. Method Sixty patients with cervical spondylotic arteriopathy were randomized, in order of admission times, to observation and control groups, 30 cases each. The observation group received three-step knife needle microinvasive operation and the control group, single suboccipital deltoid muscle lysis. The therapeutic effects were compared between the two groups of patients. Result In the two groups of patients the score changed significantly at the end of and three months after treatment compared with before and had a tendency towards gradual increase (P<0.05). The score difference values between pre-treatment and post-treatment and between post-treatment and three-month follow-up were significantly higher in the observation group than in the control group (P<0.05). Repeated measurement analysis integrogram showed significant differences in the scores between the two groups. The total efficacy rate was significantly higher in the observation group (96.7%) than in the control group (76.7%) (P<0.05). Conclusion Knife needle microinvasive operation has clinically a marked therapeutic effect on cervical spondylotic arteriopathy.

Three-step Method of Treatment Based on Syndrome Differentiation for Primary Bile Reflux Gastritis / 中国中医药信息杂志

Based on the records of ancient medical books, this article analyzed the etiology and pathogenesis of primary bile reflux gastritis. Combined with the clinical diagnosis and treatment experience and thinking, this article explained from various aspects of the principle, method, formula, and medicine, and divided the treatment into three steps, with a purpose to provide references for clinical treatment.