Objective To investigate the experimental methods of isolation ,culture and identification of osteoblasts from neo-natal New Zealand rabbits in vitro .Methods Two-step enzymatic digestion was adopted to isolate osteoblasts from skull tissue of neonatal New Zealand rabbits to conduct primary cultured .Inverted phase contrast microscope was employed to study the cellular morphology ,acridine orange fluorescent staining was used to detect the cell adhesion function ,methyl thiazolyl tetrazolium(MTT) assay was employed to measure their proliferation ,and Alizarin red and tetracycline staining were used to test their mineralization . Results Primary osteoblasts were successfully obtained .Inverted phase contrast microscopy showed non-adherent cells were round ,while adherent cells were irregular fusiform ,triangular or polygonal .Acridine orange staining showed the nuclei of osteo-blasts green fluorescence ,with good adhesion ability .Good mineralization ability was also demonstrated by tetracycline and alizarin red staining .Osteoblasts possessed good proliferation activity .Conclusion Utilization of two-step enzymatic digestion contributes to getting a lot of osteoblasts with typical morphological features and biological activity in a short time .

Objective To compare the clinical efficacy between laparoscopic suture and extraperitoneal suture for laparoscopic trocar incision with a length≥10 mm.Methods A total of 138 patients with gynecological surgery under laparoscopy admitted to our hospital from March 2017 to March 2023 were divided into two groups according to their admission time:53 cases in the traditional group(after laparoscopic surgery,the skin and subcutaneous tissue were sutured outside the peritoneum to close the trocar incision 10-12 mm in length)from March 2017 to March 2020,and 85 cases in the improved group(the fascia and peritoneum were sutured intraperitoneally under laparoscopy to close the trocar incision 10-12 mm in length)from April 2020 to March 2023.The postoperative incision complications were compared between the two groups.Results There was 1 case of postoperative incision liquefaction in the improved group,without other complications.There were 1 case of postoperative incision bleeding,2 cases of incision infection,1 case of incision liquefaction,1 case of incision dehiscence,and 1 case of incision hernia in the traditional group.The incidence of postoperative incision complications in the improved group was 1.2％(1/85),significantly lower than that in the traditional group[11.3％(6/53);x2=5.029,P=0.025].Conclusion After gynecological laparoscopic surgery,intra-abdominal suture of incision≥10 mm in length significantly reduces the incidence of postoperative incision complications,but the operation is difficult to perform and requires high suturing techniques.