The temporomandibular joint (TMJ) is surrounded by a joint capsule, the smooth inner layer of which is called the synovial membrane. Synovium is involved in various intraarticular diseases, and it is a key area of joint disease. Synovial type-A cells are located in the lining layer of the synovial membrane, mostly on the side of the membrane close to the joint cavity. They have a strong phagocytic effect, and their main role is to remove the degradation products of the intra-articular and extracellular matrix. Various intra-articular diseases will affect the synovium, which is the key area of joint disease. The method of cell culture in vitro can effectively simulate the growth environment of cells in vivo and can accurately understand the effects of single and multiple factors on synovial cells, which has become a basic research method. In this review paper, the latest research progress in human temporomandibular joint type-A synoviocytes is reviewed from the aspects of cell origin, in vitro culture, cell purification, and cell biological function.

Objective: To establish a Tohoku hospital pediatrics-1 (THP-1) cell line with G protein-coupled recep- tor108 ( GPR108) deletion and explore its functions． Methods: According to the requirements of the clustered regularly interspaced short palindromic repeats ( CRISPR) / CRISPR-associated protein 9 ( Cas9 ) system ，two single guide RNAs (sgRNA1 and sgRNA2) paring to the flanking fragments of human GPR108 gene were designed and synthesized．The two oligonucleotides were inserted in the pL-CRISPR. EFS．GFP vector to generate the new recombinant vectors ( pL-CRISPR. EFS．GFP-sgRNA1 and pL-CRISPR. EFS．GFP-sgRNA2 ) ．The recombinant vectors and packaging plasmids (pMD2. G and psPAX2) ，were co-transfected into 293T cells to generate virus for infecting THP-1 cells．The GFP + cells were screened and isolated in 96-well culture plates by flow cytometry to obtain single-cell clones．PCR and Western blot were used to detect whether GPR108 was successfully knocked out in THP- 1 cells．Both GPR108 + / + and GPR108 －/ － THP-1 cells were treated with lipopolysaccharide (LPS) ．Interleukin 8 (IL-8) derived from the THP-1 cells，which were treated by LPS，was detected with Western blot and cytometric bead array ( CBA) analysis. Results: The recombinant lentiviral vector pL-CRISPR. EFS．GFP-sgRNA was successfully constructed and single-cell clone F9 was obtained by flow cytometric sorting after transfection of THP-1 cells．PCR and Western blot both confirmed that F9 was a GPR108 －/ － THP-1 single-cell clone． LPS stimulated GPR108 －/ － and GPR108 + / + THP-1 cells，both Western blot and CBA results showed a significant decrease in IL- 8 synthesis and secretion in GPR108 －/ － THP-1 cells． Conclusion The GPR108 －/ － THP-1 cell clone is success- fully obtained based on the CRISPR / Cas9 system．GPR108 deletion in THP-1 cells treated by LPS leads to a decrease of IL-8 expression and secretion．It lays the foundation for further research on the molecular mechanisms of GPR108 in the immune inflammatory response.

Objective:To explore the clinical effects of penile ventral Z-shaped cross flap and penile dorsal pedicled transfer flap in penoplasty for concealed penis.Methods:From January 2017 to June 2019, the data of 151 patients with concealed penis admitted to our hospital was retrospectively reviewed. The patients were divided into 2 groups according to the surgical method. 69 cases were treated with penis ventral Z-shaped cross flap to form penis and 82 cases were treated with dorsal penis pedicled flaps to form the penis. In Z-shaped flap group, the penis length of 33 patients with tight scrotum was (3.06±0.25)cm before surgery and the penis length of 36 patients with relaxed scrotum was (2.99±0.28) cm before surgery. In flap with transfer group, the penis length of 39 patients with tight scrotum was (3.04±0.30)cm before surgery and the penis of 43 patients with relaxed scrotum was (3.04±0.24)cm before surgery. The length of the penis after surgery and incidence of postoperative complications were compared between Z-shaped flap group and flap with transfer group. Common complications included penile body retraction, foreskin edema, foreskin stenosis and penile wound splitting.Results:151 patients were followed up for 6-12 months, and all patients were satisfied with penis appearance. There was no penile necrosis or urinary fistula. In Z-shaped flap cross group, the penis length of 33 patients with tight scrotum extended (2.47±0.22)cm after surgery.The penis length of 36 patients with scrotum relaxation extended (2.61±0.27)cm after surgery, 39 patients was adopted the penile dorsal pedicled transfer flap with scrotum tight had extended penis (2.90± 0.16)cm, which significantly different from the Z-shaped flap group( P<0.05). In flap with transfer group, 43 patients with relaxed scrotum extended (2.79±0.18)cm after surgery, which was significantly different from the Z-shaped flap group ( P<0.05). In Z-shaped flap group, 33 patients with scrotum tight, there were 2 cases of penile retraction, 1 case of stenosis of the foreskin, 2 cases of foreskin edema, 2 cases of penile wound rupture. In transfer flap group, of 39 patients with scrotum tight, there was 1 case of foreskin edema. The incidence of complications that adopted the penile dorsal pedicled transfer flap with scrotum tight was lower than those adopted penile ventral Z-shaped cross flap [2.56%(1/39) vs. 21.21%(7/33), P=0.033]. In transfer flap group, of the 43 patients with scrotum relaxation, there were 3 cases of penile retraction, 3 cases of foreskin stenosis, 2 cases of penile ventral foreskin edema, and 1 case of penile wound rupture. Z-shaped flap group: 36 patients was scrotum relaxation was 1 case of foreskin edema. The incidence of complications that adopted the penile dorsal pedicled transfer flap was higer than those adopted penile ventral Z-shaped flap [20.93%(9/43) vs. 2.78%(1/36), P =0.038]. Conclusions:In terms of children with tight scrotum or loose scrotum, the effect of the transfer flap method to extend the penis is better than that of the Z-shaped flap method. However, the transfer flap method has a low complication rate for children with tight scrotum, while the Z-shaped flap method has a low complication rate for children with loose scrotum.

Fibrous hamartoma of infancy (FHI) in the scrotum of children is a rare benign soft tissue tumor, which mostly occurs in children under 2 years old. It grows rapidly in the early stage and is easily misdiagnosed as a malignant tumor adjacent to the testis. A case of FHI in the scrotum was admitted in our hospital in recent years, a tumor resection with preservation of testicle was performed, the lesion was completely removed. Postoperative follow-up was 20 months, and there was no evidence revealing recurrence of the tumor after excision.

Objective:To compare the advantages and disadvantages of laparoscopic robot-assisted transmesenteric approach and retrocolic approach disconnected pyeloplasty in the treatment of children with hydronephrosis.Methods:From October 2020 to March 2021, 19 children with hydronephrosis were divided into two groups: intra-renal type and extra-renal type. Among them, 15 were males and 4 were females. The average age of the patients was 3.5 years old (0.2 years old to 16.8 years old), and the average weight was 18.4 kg (5.5 kg to 67.0 kg). The average ERPF of affected kidney before surgery was 35.4%(23.0%-49.8%). All of them were treated with laparoscopic robot-assisted transmesenteric approach and retrocolic approach disconnected pyeloplasty. The operation was performed in accordance with the standard surgical procedures of the guidelines. After the insertion of the trocar, the children in the transmesenteric group were exposed to the renal pelvis by incising the colonic mesangium into the retroperitoneal space, while in the retrocolic group, the peritoneum was cut into the retroperitoneal space to expose the renal pelvis. After that, the steps of incision, cutting, tube placement, and suture of the renal pelvis and ureter were the same in the two groups. Among the 10 cases of the extrarenal type, 6 cases were in the transmesenteric group and 4 cases were in the retrocolic group; among the 9 cases of the intrarenal type, 5 cases were in the transmesenteric group and 4 were in the retrocolic group. There was no statistically significant difference in age, weight, and renal function of the affected side before operation in different surgical approach groups ( P>0.05). The operation time, intraoperative anastomosis time, intraoperative blood loss and postoperative hospital stay were recorded and compared. There was no statistical difference in the age, weight, and renal function of the affected side before the operation. Results:19 cases were followed up for 6 months, no complications such as fever or wound infection occurred. The operation was successfully completed in all patients, no patients were transferred to open surgery, and the hydronephrosis was significantly reduced. Symptoms disappeared in both groups. Of the 19 children. In children with extrarenal type, the operation time of the transmesenteric group and the retrocolic group were (108.8±15.5) min and (132.8±7.6) min, and the intraoperative anastomosis time was (40.7±6.1) min and (51.5±5.5)min, the estimated intraoperative blood loss was (9.5±2.1) ml and (9.3±0.8) ml, respectively, and the postoperative hospital stay was (9.0±1.6) d and (9.3±2.9) d. The operation time and the difference of intraoperative anastomosis time was statistically significant ( P<0.05). In children with intrarenal type, the operation time of the transmesenteric group and the retrocolic group were (136.6±7.9) min and (116.5±13.5) min, and the intraoperative anastomosis time was (52.8±6.9) min and (40.8±6.2), min, the estimated blood loss during the operation was (11.4±2.3) ml and (10.5±0.9) ml, and the postoperative hospital stay was (8.8±1.7) d and (8.0±1.6) d. The operation time and The difference of intraoperative anastomosis time was statistically significant ( P<0.05). The 19 cases were followed up for 6 months, and there was no complications such as fever or wound infection. The volume of hydronephrosis was significantly reduced compared with that before operation, and the renal blood perfusion increased compared with that before operation. The difference was statistically significant ( P<0.05). Conclusion:In terms of shortening the operation time and suture time, for laparoscopic robot-assisted transmesenteric approach and retrocolic approach disconnected pyeloplasty in the treatment of children with hydronephrosis, the transtransmesenteric approach is more advantageous in the treatment of extrarenal hydronephrosis, while the retrocolic approach is more advantageous in the treatment of intrarenal hydronephrosis.

Objective To investigate the effect of L-carnitine (LCNT) combined with epirubicin (EPI) on cell proliferation and apoptosis of two lung adenocarcinoma cell lines (GLC-82 and A549). Methods GLC-82 and A549 cells were divided into control group, EPI group, LCNT group and EPI+LCNT group, respectively. The cell proliferation rate was examined by MTT assay 24 h after the treatment, and the cell cycle and cell early apoptosis were analyzed by flow cytometry. The expression of P53 and Bcl-2 proteins were detected by Western Blot. Results Compared with the control group, the proliferation rate of GLC-82 and A549 cells was 37.56％(P<0.01) and 6.32％(P>0.05) after 24 hours of 20.00μg/ml LCNT treatment indicating LCNT could promote the proliferation of GLC-82 cells. Compared with the EPI group, EPI+LCNT had smaller inhibitory effect on the proliferation of GLC-82 cells, and the inhibition rate of the EPI+LCNT group was 12.14％lower than that of EPI group (P<0.05). Compared with the EPI group, EPI+LCNT could significantly increase the G0/G1 phase ratio of GLC-82 cells (50.42％±1.21％vs. 25.94％± 0.66％, P<0.01) and A549 cells (54.92％±1.71％vs. 38.63％±0.69％, P<0.01), decrease the S phase ratio of GLC-82 cells (34.21％±0.96％vs. 59.68％±1.25％, P<0.05), and prevent the early apoptosis of GLC-82 cells without affecting apoptosis of A549 cells. Moreover, in the EPI+LCNT group, the expression of P53 protein in GLC-82 and A549 cells was down-regulated (P<0.05), and the expression of Bcl-2 protein in GLC-82 cells was up-regulated (P<0.01) and no significant change in A549 cells (P>0.05). Conclusions Comparing with the EPI, the combination of LCNT and EPI has less proliferation inhibition and apoptosis induction on GLC-82 cells, and without significant effect on A549 cells. LCNT can promote the proliferation of GLC-82 cells and block the cell cycle at G0/G1 phase. This mechanism may be related to down-regulation of P53 protein and up-regulation of Bcl-2 protein expression.

Objective To evaluate Sysmex XS-800i,XS-1000i and XE-2100D hematology analyzers when used to detect RBC count,hemoglobin (HGB),hematocrit (HCT),mean corpuscular volume (MCV),mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC).Methods XE-2100D hematology analyzer was calibrated after performance evaluation,and the three analyzers had the intra-day precisions measured with three levels of whole-blood quality control materials.Totally 50 whole-blood specimens were detected with the three analyzers respectively,and statistical analyses and clinically acceptable performance evaluation were carried out on RBC count and the obtained results.Results XE-2100D hematology analyzer met the clinical requirements,and the three analyzers all gained high precisions when used to measure the parameters of the whole-blood quality control materials.The correlation coefficients (r2) respectively between the three analyzers were all higher than 0.95 when used to test the 50 specimens.At all medical decision levels XS-800i and XS-1000i hematology analyzers both gained acceptable detection results except XS-800i hematology analyzer in case of 5.9×1012/L RBC count as well as 35％ or 50％ HCT.Conclusion Sysmex XS-800i,XS-1000i and XE-2100D hematology analyzers have high precisions and correlations when used to detect RBC count,HGB,HCT and MCV,and contrast test is suggested to be executed periodically to ensure the comparability of tbe result.

OBJECTIVE:To study the inhibitory effect of dihydrotanshinone(DTS)on human lung cancer GLC-82 cell and its mechanism. METHODS:After treated with 0(blank control),5,10,20,40,80 and 100 μg/ml DTS for 24 and 48 h,MTT as-say was used to measure the inhibition rates and IC50 of cells;cell apoptosis was detected by flow cytometry after treated with 17.85 μg/ml DTS for 12,24 and 48 h to calculate apoptotic rate;Western blot was used to detect the protein expressions of Bcl-2, Bax and Caspase-3. RESULTS:Compared with blank control group,different concentrations of DTS inhibited the proliferation of cells;24 and 48 h maximal inhibition rate were 54.48% and 64.95%,respectively;IC50 were 62.36 and 33.94 μg/ml. DTS could induce cell apoptosis in positive time dependent manner,and the range of inhibition rate was 5.6%-29.6%;Western blot showed DTS could down-regulate the expression of Bcl-2 protein and up-regulate the expression of Caspase-3 protein (P<0.01 or P<0.05). CONCLUSIONS:DTS have significant inhibitory effect on GLC-82 cells and also induce cell apoptosis,by a possible mech-anism of down-regulating the expression of Bcl-2 protein and up-regulating the expression of Caspase-3 protein.

Tumor immunotherapy is highlighted in tumor studies and has a remarkable curative effect on tumors. Programmed cell death-1 (PD-1), which is mainly expressed in activated T and B cells, is an important immune inhibitory molecule. Its ligand PD-L1 could be highly expressed in cancer cells, and the PD-1 pathway shows sustained activation in a tumor micro-environment. PD-1/PD-L1 inhibitors can block the combination of PD-1 and PD-L1, inhibit negative to regulatory signals, and restore the activity of T cells, thereby enhancing immune response. Recent studies showed that PD-1 and PD-L1 inhibitors have been effective in many tumor types. This review summarizes PD-1/PD-L1 inhibitors and their clinical effects on different tumor types.

OBJECTIVETo investigate the profile of promoter methylation and expression of SHP-1 gene in the progression of chronic myeloid leukemia (CML).

METHODSThe expression level of SHP-1 mRNA and protein in bone marrow or peripheral blood mononuclear cells from CML patients were detected by Western blot and SYBR Green-based qRT-PCR. The methylation status of SHP-1 were assessed by methylation-specific polymerase chain reaction (MSP) assay. K562 cells were infected with the lentiviral plasmids pEX-SHP-1-puro-Lv105 (K562-SHP-1) or pEX-EGFP-puro-Lv105 (K562-EGFP). The levels of proteins and phosphorylated proteins were detected by Western blot. qRT-PCR assay was used to test the level of BCR-ABL mRNA.

RESULTSThe relative levels of SHP-1 mRNA were sharply decreased in advanced stages CML compared to chronic phase (CP)-CML (0.79±0.37 vs 1.18±0.64, P= 0.009). The level of SHP-1 protein was lower in advanced stages CML compared to CP-CML (0.57±0.02 vs 1.02±0.04, P=0.039). The frequency of SHP-1 gene promoter methylation at selected loci in CP-CML was 23.8% (10/42), and the methylated regions were detected in all advanced CML samples (P<0.01). SHP-1 was stably transfected into K562 cells and selected with puromycin. Overexpression of SHP-1 inhibited the proliferation and induced the apoptosis of K562 cells, meanwhile leaded to G0/G1 phase arrest. After transfection, the level of BCR-ABL mRNA was not affected in K562-SHP-1 cells (1.32±0.34) compared to K562-EGFP cells (1.18±0.20, P=0.644), but overexpression of SHP-1 caused a slight decrease in BCR-ABL protein in K562-SHP-1 cells compared to K562 -EGFP cells (0.78±0.15 vs 1.27±0.24, P=0.040). Overexpression of SHP-1 resulted in a remarkable decrease in MYC protein, phosphorylated forms of JAK2, STAT5, Akt and MAPK. However, the un-phosphorylated forms of these molecules were not significantly affected.

CONCLUSIONDecreased expression of SHP-1 caused by aberrant promoter hypermethylation may play a key role in the progression of CML by dysregulation of BCR-ABL, Akt, MAPK, MYC, JAK2 and STAT5 signaling.

Apoptosis ; DNA Methylation ; Disease Progression ; Fusion Proteins, bcr-abl ; Humans ; Janus Kinase 2 ; K562 Cells ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; Leukocytes, Mononuclear ; Phosphorylation ; Protein Tyrosine Phosphatase, Non-Receptor Type 6 ; RNA, Messenger