Objective To investigate the destructive effect of CSC-DC-CIK who were induced by cytokine induced killer (CIK) cells co-cultured with dendritic cells (DCs) on homologous tumor cells and to explore the possibility of CSC anti-gen involving in killing tumor. Methods Kidney cancer stem cells (KSCs) and lung cancer stem cells (LSCs) were isolated through FACS using CD133 +as a selection marker from cultured kidney cancer cell line A498 and lung cancer cell line A549 respectively. Freeze-thaw method was used to obtain the cancer stem cells(CSCs)antigens. DC cells and CIK cells were collected by in vitro expansion and inducted from the mononuclear cells isolated from human cord blood. The CIK cells were co-cultured with the DCs which were pulsed with the CSCs antigens(CSC-DC-CIK)mentioned above. Immunopheno-types of DC and CIK were analyzed by flow cytometry;cytokines levels were detected by ELISA kits and the destructive ef-fects of two kinds of CSC-DC-CIKs were tested by lactate dehydrogenase (LDH) release assay. Results The expression of phenotypes CD40+, CD80+, CD86+and HLA-DR+were higher in CSC-DC than in CD(P<0.01);the expression of pheno-types CD40+, CD80+, CD86+and HLA-DR+of DC and CSC-DC were higher after co-culture than those before co-culture( P<0.01);the expression of phenotypes CD40+, CD80+, CD86+and HLA-DR+of CSC-DC after been co-cultured with CIK were higher than those of DC after been co-cultured with CIK(P<0.01). The CIK phenotypes:CD3+, CD8+, CD56+were in-creased in CIK co-cultured with both CSC-DC and DC than those before co-culture (P<0.01);the expression of pheno-types CD3+, CD8+, CD56 +were higher in CSC-DC co-cultured with CIK than in DC co-cultured with CIK. DC-CIK and CSC-DC-CIK groups were more capable to express IFN-γ, TNF-α, IL-2 than they were before co-cultured with CIK (P<0.01). CSC-DC-CIK group can secrete more above cytokines than DC-CIK group does(P<0.01). The destructive rates of KSC-DC-CIK and LSC-DC-CIK on target cells were (50.21 ± 4.24)%and (49.32 ± 3.89)%respectively which were much higher than that in DC-CIK(30.25±3.11)%(F=89.157,P<0.01). Conclusion CSC-DC-CIKs have destructive effects on homologous tumor cells. More researches are needed to explore the mechanism and to evaluate the clinical applications.

Objective To examine the effect of positive and negative evaluative conditioning (EC) on neutral faces.Methods The experiment consisted of three phases:baseline phase,conditioning phase,and re-evaluative phase,in which 41 college students participated,watching sequences of neutral faces (CS) pairing to either a positive stimulus (USpos) or a negative stimulus (USneg).Their emotional experiences (valence and arousal) and physiological reactivity (eyeblink startle reflex and skin conductance) were recor ded.Results (1) In the re-evaluative phase,CSpos was rated significantly more positive than CSneg and CSneut,while CSneg was rated significantly more negative than CSneut (CSpos (5.05± 1.24),CSneg (3.73± 1.48),CSneut (4.46± 1.04),P＜0.05).(2)In the re-evaluative phase,the mean startle eyeblink response magnitude(T score) to CSpos was significantly smaller than the responses elicited by CSneg and CSneut (CSpos (45.04±5.56),CSneg (51.44±9.30),CSneut (54.52± 10.60),P＜0.01).Conclusion The findings suggest that neutral faces can acquire valences and approach motivation through EC.

Objective To establish characteristic parameters of plural impedance spectra of rabbit whole blood cell.Method The AC impedances of 30 blood samples from the 10 rabbits have been measured with the Agilent 4294A impedance analyzer at frequency range of 0.01～100MHz,then its characteristic data have been analyzed by the Bode plot,the Nyquist plot and the Nichols plot.Results(1)The impedance amplitude and phase angle of rabbit blood cell have a dependence of frequency.(2)The impedance spetroctra of rabbit blood cell have two characteristic frequency:the characteristic 1st frequency(fC1) was 2.58MHz,the characteristic 2nd frequency(fC2) was 5.21MHz.Conclusion The frequency properties of blood cells can be obtained by the analysis of impedance spectroscopy.

Objective To investigate the effect of verapamil on chemosensitivity and apoptosis of esophageal cancer cells.Methods The IC50 values (IC501) of three kinds of chemotherapy drugs:paclitaxel,cisplatin,and 5-fluorouracil were detected by CCK-8 method in 4 kinds of esopha geal cancer cell lines (EC109,EC9706,KYSE-450,TE-1).Each cell was processed by verapamil (4.91 μg/mL) combined with the chemotherapy treatment above,IC50 (IC502) value was detected by CCK8 assay combined with chemotherapy for the verapamil of the 4 esophageal carcinoma cell lines (EC109,EC9706,KYSE-450 and TE-1).A pair of esophageal cancer cells with the most significant reversal of the resistance to chemotherapy by VER detected by the previous experiments were included as research objects,and Annexin V-PI double staining method was used to detect the apoptosis of esophageal cancer cells.Results The IC50 values of paclitaxel in KYSE-450 and EC9706 cell lines were significantly higher than that in EC109 and TE-1.The IC50 values of cisplatin in TE-1 cells were significantly less than those in the other three cells,while the IC50 values of 5-fluorouracil in EC109 were significantly higher than that in other three cell lines.After adding verapamil,the IC50 values of paclitaxel,cisplatin and 5-fluorouracil in four esophageal cancer cell lines (EC109,EC9706,KYSE-450 and TE-1) all decreased to different degrees,and the above three kinds of chemotherapeutic drugs had increased their sensitivity to varying degrees.Among them,the cisplatin group before adding verapamil had different degrees of resistance compared with EC109,EC9706,KYSE-450,TE-1.After joining verapamil,IC50 value change for the EC9706 multiplied for 15.2 times,suggesting that the reversal was sensitive;IC50 value change for EC109 was 1.6 times smaller,suggesting that the reversal was tolerant.Conclusion Verapamil reversal of EC9706 adriamycin has highest efficiency index (15.2 times),there was significant difference compared with the EC109 cells (1.6 times).Tipthe EC9706 (DDP) sensitivity to the reversal of verapamil,and EC109 (DDP) reverse resistance of verapamil.Verapamil may enhance the ability to reverse chemotherapy tolerance by promoting cell apoptosis,thereby enhancing the sensitivity of chemotherapeutic drugs.

Objective To investigate the effect of verapamil on chemosensitivity and apoptosis of esophageal cancer cells.Methods The IC50 values (IC501) of three kinds of chemotherapy drugs:paclitaxel,cisplatin,and 5-fluorouracil were detected by CCK-8 method in 4 kinds of esopha geal cancer cell lines (EC109,EC9706,KYSE-450,TE-1).Each cell was processed by verapamil (4.91 μg/mL) combined with the chemotherapy treatment above,IC50 (IC502) value was detected by CCK8 assay combined with chemotherapy for the verapamil of the 4 esophageal carcinoma cell lines (EC109,EC9706,KYSE-450 and TE-1).A pair of esophageal cancer cells with the most significant reversal of the resistance to chemotherapy by VER detected by the previous experiments were included as research objects,and Annexin V-PI double staining method was used to detect the apoptosis of esophageal cancer cells.Results The IC50 values of paclitaxel in KYSE-450 and EC9706 cell lines were significantly higher than that in EC109 and TE-1.The IC50 values of cisplatin in TE-1 cells were significantly less than those in the other three cells,while the IC50 values of 5-fluorouracil in EC109 were significantly higher than that in other three cell lines.After adding verapamil,the IC50 values of paclitaxel,cisplatin and 5-fluorouracil in four esophageal cancer cell lines (EC109,EC9706,KYSE-450 and TE-1) all decreased to different degrees,and the above three kinds of chemotherapeutic drugs had increased their sensitivity to varying degrees.Among them,the cisplatin group before adding verapamil had different degrees of resistance compared with EC109,EC9706,KYSE-450,TE-1.After joining verapamil,IC50 value change for the EC9706 multiplied for 15.2 times,suggesting that the reversal was sensitive;IC50 value change for EC109 was 1.6 times smaller,suggesting that the reversal was tolerant.Conclusion Verapamil reversal of EC9706 adriamycin has highest efficiency index (15.2 times),there was significant difference compared with the EC109 cells (1.6 times).Tipthe EC9706 (DDP) sensitivity to the reversal of verapamil,and EC109 (DDP) reverse resistance of verapamil.Verapamil may enhance the ability to reverse chemotherapy tolerance by promoting cell apoptosis,thereby enhancing the sensitivity of chemotherapeutic drugs.

BACKGROUND:Mechanical stimulation has been confirmed to promote osteogenic differentiation of bone marrow stromal stem cells,but the mechanism is unknown.Primary cilia are important mechanoreceptors and regulate various signaling pathways such as TGF-β1/BMP-2/SMAD.They are likely to be important targets for mechanical regulation of bone marrow stromal stem cells. OBJECTIVE:To investigate the effect and mechanism of fluid shear stress on osteogenic differentiation of bone marrow stromal stem cells. METHODS:Rat bone marrow stromal stem cells were divided into control group,mechanical stimulation group(fluid shear mechanics intervention by shaking table),mechanical stimulation + IFT88 silencing group(mechanical stimulation + silencing IFT88 expression with siRNA).After 24 hours of intervention,qRT-PCR was utilized to determine the expression of transforming growth factor β1 and bone morphogenetic protein 2.Western blot assay was used to detect the expression of phosphorylated SMAD2/3 protein.Immunofluorescent staining of primary cilia was conducted and morphology was analyzed. RESULTS AND CONCLUSION:Shear stress stimulation could promote the transcriptional activity of transforming growth factor β1 and bone morphogenetic protein 2 genes,and increase the expression of phosphorylated SMAD2/3 protein.After siRNA interfered with primary cilia,this mechanical response effect was significantly reduced.There was a Spearman correlation between the change ratio of the primary cilium area of bone marrow stromal stem cells and the increased ratio of transforming growth factor β1 and bone morphogenetic protein 2 gene transcription.These findings indicate that primary cilia/intraflagellar transport mediates the activation of fluid shear stress-responsive transforming growth factor β1/bone morphogenetic protein 2/SMAD signaling pathway and promotes osteogenic differentiation of bone marrow stromal stem cells.

Objective:To investigate the perinatal prognosis and its impact factors for premature infants with twin-twin transfusion syndrome (TTTS) who were born at ≤34 weeks of gestation.Methods:A retrospective study was conducted on 68 pregnancies of TTTS with gestational age ≤34 weeks at delivery, among them 106 preterm infants (TTTS group) were admitted to the neonatal intensive care unit of the First Affiliated Hospital, Sun Yat-sen University from January 2003 to February 2019. During the same period, another 178 twins without TTTS, congenital malformation, and intrauterine intervention who matched the TTTS group in maternal age (differences within two years) and gestational age (differences within one week) were assigned as non-TTTS group. Perinatal prognosis of TTTS infants born at ≤34 weeks was analyzed by comparing the differences in postnatal early complications and perinatal outcomes (survival time morn than 28 days or not) between the TTTS and non-TTTS groups, recipient and donor twins, mild and severe TTTS infants, and among TTTS infants with different intrauterine interventions. The risk factors for perinatal survival in TTTS infants with gestational age ≤34 weeks were analyzed. Two independent samples t-test, one-way analysis of variance, rank-sum test, Chi-square test, and ordered logistic regression were used for statistical analysis. Results:(1) Among the 68 pregnancies, the overall perinatal survival rate of the neonates was 72.1% (98/136), the double-twin survival rate was 48.5% (33/68), and the rate of at least one survivor was 95.6% (65/68). (2) In the TTTS group, 62 were recipients and 44 were donors. Stage Ⅰ-Ⅱ TTTS was found in 41 cases (mild TTTS group) and stage Ⅲ-Ⅴ in 65 cases (severe TTTS group). (3) The rate of severe brain injury was higher in the severe-TTTS group than those in the mild-TTTS group [9.2% (6/65) vs. 0.0% (0/41), χ 2=4.01, P=0.045]. (4) Gestational age ≤28 weeks ( OR=101.90, 95% CI: 5.07-2 048.37), stage Ⅳ ( OR=14.04, 95% CI: 1.56-126.32) and stage Ⅴ TTTS ( OR=51.09, 95% CI: 3.58-728.81) were independent risk factors for death within 28 days (all P<0.05). (5) Compared with the non-TTTS group, the TTTS group had higher rates of neonatal anemia [51.9% (55/106) vs. 33.1% (59/178), χ 2=9.71], polycythemia [5.7% (6/106) vs. 0.6% (1/178), χ 2=7.18], neonatal persistent pulmonary hypertension [3.8% (4/106) vs. 0.0% (0/178), χ 2=6.81], sepsis [15.1% (16/106) vs. 7.3% (13/178), χ 2=4.40], state Ⅲ or higher retinopathy of prematurity [3.8% (4/106) vs. 0.0% (0/178), χ 2=6.81], congenital cardiac structural abnormality [19.8% (21/106) vs. 0.6% (1/178), χ 2=33.45], heart failure [8.5% (9/106) vs. 0.6% (1/178), χ 2=12.29], and renal insufficiency [14.2% (15/106) vs. 1.1% (2/178), χ 2=20.04] (all P<0.05). Conclusions:Compared with the twin premature infants without TTTS, those with TTTS and ≤34 gestational age were more likely to have cardiac, cerebral, and renal complications. The more severe the TTTS, the higher the incidence of severe brain injury. TTTS preterm infants with gestational age ≤28 weeks and stage Ⅳ or above have high risk of death.

BACKGROUND:Traditional surgery for lumbar disc herniation involves extensive excision of tissue surrounding the nerve for decompression and removal of protruding lumbar intervertebral discs,which poses various risks and complications such as nerve damage causing paralysis,lumbar instability,herniation recurrence,intervertebral space infection,and adjacent vertebral diseases. OBJECTIVE:To propose the symmetrically adduction of lumbar decompression induced resorption of herniated nucleus pulpous technique for lumbar spine symmetrically decompression,showing the induced resorption of herniated nucleus pulpous phenomenon and early clinical efficacy,and then analyze its decompression mechanism. METHODS:214 patients with lumbar disc herniation at Department of Orthopedics,First Affiliated Hospital of Zhengzhou University from March 2021 to May 2023 were enrolled in this study.Among them,81 patients received conservative treatment as the control group,and 133 patients received symmetrically adduction of lumbar decompression induced resorption of herniated nucleus pulpous treatment as the trial group.Before surgery,immediately after surgery(7-14 days),and early after surgery(over 1 year),MRI images were used to measure the volume changes of lumbar disc herniation.CT images were used to measure the posterior displacement distance of the lumbar spinous process ligament complex,as well as the width and height of the lateral recess.Japanese Orthopaedic Association scores were used to evaluate the patient's neurological function recovery. RESULTS AND CONCLUSION:(1)Control group:81 patients with lumbar disc herniation were treated conservatively,with a total of 171 herniated lumbar discs.The average follow-up time was(22.7±23.1)months.The first and second MRI measurements of 171 herniated lumbar discs showed herniated lumbar disc volumes of(551.6±257.9)mm3 and(792.2±330.4)mm3,respectively,with an average volume increase rate of(53.2±44.4)%,showing statistically significant differences(P<0.001).Out of 171 herniated lumbar discs,4 experienced natural shrinkage,with an absorption ratio of 2.3%(4/171)and an absorption rate of(24.5±9.9)%.(2)Trial group:133 patients with lumbar disc herniation had a total of 285 herniated lumbar discs.(1)Immediately after surgery:All patients were followed up immediately after surgery.229 out of 285 herniated lumbar discs experienced retraction,with an absorption ratio of 80.3%(229/285)and an average absorption rate of(21.5±20.9)%,with significant and complete absorption accounting for 6.5%.There were a total of 70 herniated lumbar discs in the upper lumbar spine,with an absorption ratio of 85.7%(60/70),an average absorption rate of(23.1±19.5)%,and a maximum absorption rate of 86.6%.There were 215 herniated lumbar discs in the lower lumbar spine,with an absorption ratio of 78.6%(169/215),an average absorption rate of(21.0±21.3)%,and a maximum absorption rate of 83.2%.Significant and complete absorption of the upper and lower lumbar vertebrae accounted for 5.7%and 6.5%,respectively,with no statistically significant difference(P>0.05).The average distance of posterior displacement of the spinous process ligament complex immediately after surgery was(5.2±2.8)mm.There were no significant differences in the width and height of the left and right lateral recess before and immediately after surgery(P>0.05).The Japanese Orthopaedic Association score immediately after surgery increased from(10.1±3.4)before surgery to(17.0±4.8),and the immediate effective rate after surgery reached 95.6%.(2)Early postoperative period:Among them,46 patients completed the early postoperative follow-up.There were 101 herniated lumbar discs,with an absorption ratio of 94%(95/101)and an average absorption rate of(36.9±23.7)%.Significant and complete absorption accounted for 30.6%,with a maximum absorption rate of 100%.Out of 101 herniated lumbar discs,3 remained unchanged in volume,with a volume invariance rate of 2.97%(3/101).Out of 101 herniated lumbar discs,3 had an increased volume of herniated lumbar discs,with an increase ratio of 2.97%(3/101)and an increase rate of(18.5±18.4)%.The Japanese Orthopaedic Association score increased from preoperative(9.3±5.1)to(23.5±4.0),with an excellent and good rate of 93.4%.(3)The early postoperative lumbar disc herniation absorption ratios of the control group and trial group were 2.3%and 85.9%,respectively,with statistically significant differences(P<0.001).(4)Complications:There were two cases of incision exudation and delayed healing in the trial group.After conservative treatment such as dressing change,no nerve injury or death occurred in the incision healing,and no cases underwent a second surgery.(5)It is concluded that symmetrically adduction of lumbar decompression induced resorption of herniated nucleus pulpous is a new method for treating lumbar disc herniation that can avoid extensive excision of the"ring"nerve and achieve satisfactory early clinical efficacy.It does not damage the lumbar facet joints or alter the basic anatomical structure of the lateral recess,fully preserves the herniated lumbar discs,and can induce significant or even complete induced resorption of herniated nucleus pulpous.Symmetrically adduction of lumbar decompression induced resorption of herniated nucleus pulpous provides a new basis and method for the clinical treatment of lumbar disc herniation.

Lymphangioleiomyomatosis (LAM) is a rare, multisystemic, low-grade neoplasm character-ized by diffuse cystic lesions in the lung.In recent years, emerging imaging examination such as ⁶⁸Ga-NEB PET-CT scan provides efficient and precise non-invasive diagnostic methods to detect lymphatic circulation abnormalities in LAM patients. The long-term efficacy and safety of sirolimus for LAM has accumulated further evidence, and genetic profiling studies have unveiled more information of genetic mechanisms. Prognosis of LAM has been much improved. We briefly reviewed the research advances of LAM in China and other countires.