Objective:To clarify the effect of voltage-gated proton channel 1 (Hv1) on the migration and invasion of breast cancer cells. Methods:The protein expression of Hv1 was detected in human breast cancer cell lines with different metastatic abilities. SiRNA technique was used to down-regulate the expression of Hvl in breast cancer MDA-MB-231 cells. Scratch and matrigel invasion methods were used to observe the effect of Hvl on the migration and invasion of breast cancer cells, and the relevant molecular mechanism was explored. Results:Hv1 was highly expressed in the highly metastatic breast cancer cell line MDA-MB-231. Hvl was more highly expressed in MDA-MB-231 cells with higher metastatic ability. The SiRNA sequence target at Hvl inhibited Hvl expression. Scratch and matrigel invasion experiments showed that the migration and invasion of MDA-MB-231 cells were significantly attenuated when Hv1 was knocked down by siRNA targeting Hv1. Zymography experiment on matrix metalloproteinase indicated that the enzyme activities of MMP-2 markedly decreased. Conclusion:Hv1 promoted the migration and invasion ability of breast cancer cells.

Objective:To analyze the clinico-pathological characteristics, pathological diagnosis, and treatment of rhabdoid tu-mor. Methods:The medical records of four rhabdoid tumor patients that were admitted to the Tianjin Medical University Cancer Insti-tute and Hospital since 2000 were analyzed based on existing literature. Results:In one of the four cases, the tumor originated from the kidney, whereas in the other three, the tumor occurred from extra-renal soft tissues. Histologic analysis revealed that the tumor cells were loosely arranged with diffuse growth, vesicular nuclei, dyed cytoplasm, visible eosinophilic inclusions, and more nuclear fission. The results of immunohistochemical staining showed that the vimentin and epithelial membrane antigen were positive, whereas CK, CD99, CD34, and S-100 were positive at different degrees. MyoD1, Desmin, and INI-1 were negative. Conclusion:Rhabdoid tumor is rare and highly aggressive. It occurs mainly in the kidney and can also be found in other systems. The unique pathological form and im-munohistochemical staining observed on the tumor can be used as reference for diagnosis.

Objective To establish a method for isolating and culturing fibroblasts from cirrhotic liver in vitro,observe the biological characteristics of fibroblasts.Methods Human fibroblasts from cirrhotic liver were isolated and cultured by adhering to the culture plastic.The morphologic and growth characteristics of the acquired cells were observed by microscopy imaging and cell counting.The expression of FSP-1 and Vimentin of fibroblasts was detected through immunofluorescence and western blotting.Results The tissue blocks were well-adherent to the culture plastic within 2 hours.Hepatocyte-like cells were observed surrounding the blocks within about 1 week,and spindle-shaped fibroblasts were observed within about 2 weeks.After a series of passage,the attached cells became proliferated quickly.The growth curves of the passage 3,4 and 5 were quite similar.The result of immunofluorescence showed the expression of FSP1 and Vimentin was positive.The positive rate was respectively (90.6 ± 1.0) ％ and (91.2-± 4.1) ％.Conclusions Human fibroblasts from cirrhotic liver can be cultured successfully through adherent property.The method for isolation and culture of fibroblasts from cirrhotic liver is convenient,efficient,stable and cultured cells remain naive biological characteristics.

Objective To investigate the role of mucosal mast cells infiltration and degranulation with nerve growth factor (NGF)in development of visceral hypersensitivity in Sprague-Dawley (SD)rats. Methods The model of visceral hypersensitivity of irritable bowel syndrome (IBS)was established in 19 neonate SD rats with intestinal stimulation (rectalballon distention)on 8th,10th and 12th postnatal days. The other 19 neonate SD rats without colonic distention were assigned to the control group.After rats grew up (six to eight weeks old),the visceral sensitivity was tested by abdominal withdrawal reflex (AWR)in 10 rats of each group.Mast cell infiltration and degranulation were observed with toluidine blue staining in colon tissue slides.The NGF level of intestinal tissues was detected by enzyme-linked immunosorbent assay (ELISA)methods in the left nine rats of each group.The culture system of dorsal root ganglias (DRG)from the neonatal rats was set up.The changes of electrophysilogical characters of DRG stimulated with NGF (100 ng/mL)for four days were recorded with patch-clamp.Paired t test was performed for comparison between groups.Results The results of AWR indicated that neonatal colonic stimulation could significantly increase visceral sensitivity after growing up.Under 20,40 and 60 mmHg (1 mmHg=0.133 kPa)distention pressure,visceral sensitivity scores of visceral hypersensitivity rats and rats of control group were 1 .00±0.50 vs 1 .67 ±0.50,1 .89 ±0.31 vs 2.89 ±0.34 and 2.89 ±0.33 vs 3.89±0.33,the differences were statistically significant (t=-2.83,-6.00 and -6.00,all P <0.05 ). The results of master cells staining in tissue slides showed colonic master cells infiltration was obvious in rats with visceral hypersensitivity,and part of mast cells were degranulation.The result of ELISA demonstrated that NGF level of visceral hypersensitivity rats was significantly higher than that of control group ((11.07±3.06)pg/mg vs (2.38 ±1.88)pg/mg,t =-6.93,P <0.05).The results of electrophysilogical tests of primary cultured DRG indicated that compared with blank control growp,the action potential threshold of neuron in NGF 100 ng/mL group significantly decreased ((-18.0±2.1 )mV vs (-29.0 ± 2.5 )mV,t = 12.26,P <0.05)and discharge frequency increased ((5 .0± 1 .4 )/800 ms vs (12.0 ± 3.2)/800 ms,t=-8.40,P <0.05 ).Meanwhile,neuron voltage-gated K+ current density remarkably decreased,most were sustained delayed rectifier K+ current (I K )decreasing ((279.0 ±48.0)pA/pF vs (203.0±39.0)pA/pF,t=6.18,P <0.05).Conclusion Colonic stimulation in neonatal rats could cause intestinal master cells infiltration and degranulation,which induced changes of neuron electrophysilogical characters and resulted in visceral hypersensitivity after growing up.

This article reported a case of fetal testicular torsion diagnosed by prenatal ultrasound. Ultrasound examination at 38 weeks of gestation showed a slightly enhanced echo in the left testicle of the fetus and fluid accumulation within the testicular sheath cavity and between the sheath and fascia, presenting a 'double-ring' sign. An emergency cesarean section was performed considering the risk of acute fetal testicular torsion. Postnatal ultrasound images revealed abnormalities in the left testicle of the neonate and peripheral effusion with a 'double-ring' echo. The patient was considered to have congenital testicular torsion (with a high possibility of extravaginal type) and received an emergency treatment of left testicular torsion reduction combined with testicular fixation surgery and right testicular fixation surgery. Six months after surgery, reexamination by ultrasound showed the testicles of the newborn were normal.

Wastewater resources, CO2 emission reduction and microalgae biodiesel are considered as current frontier fields of energy and environmental researches. In this paper, we reviewed the progress in system of microalgae culture for biodiesel production by wastewater and stack gas. Multiple factors including microalgal species, nutrition, culture methods and photobioreactor, which were crucial to the cultivation of microalgae for biodiesel production, were discussed in detail. A valuable culture system of microalgae for biodiesel production or other high value products combined with the treatment of wastewater by microalgae was put forward through the optimizations of algal species and culture technology. The culture system coupled with the treatment of wastewater, the reduction of CO2 emission with the cultivation of microalgae for biodiesel production will reduce the production cost of microalgal biofuel production and the treatment cost of wastewater simultaneously. Therefore, it would be a promising technology with important environmental value, social value and economic value to combine the treatment of wastewater with the cultivation of microalgae for biodiesel production.
Biodegradation, Environmental ; Biofuels ; Biotechnology ; methods ; Carbon Dioxide ; metabolism ; Cell Culture Techniques ; methods ; Cells, Cultured ; Microalgae ; growth & development ; metabolism ; Photobioreactors ; microbiology ; Waste Disposal, Fluid ; methods ; Waste Water ; microbiology

Objective: To detect the expression of SVEP1, PKHD1 and P53 in primary liver cancer tissues by immunohistochemistry for predicting the recurrence of liver cancer. Methods: The clinical data of 103 patients with primary liver cancer who underwent surgical resection at Tianjin Medical University Cancer Institute and Hospital were gathered from January 2013 to January 2014 and analyzed retrospectively. Expression values of three different proteins were used to develop separate immunohistochemical scores for the prog-nosis of recurrence in patients. The patients were classified into either a high-risk or a low-risk group based on their immunohisto-chemical scores through ROC curve analysis. The difference in recurrence ratio between the two groups was then compared using the common research index of disease-free survival (DFS). Results: The median age of the total patients was 55 years (range 21-88 years), the median AFP level was 70.6 (range 1.03-718840.0) μg/L, the median CA19-9 level was 22.89 (range 0.6-1000.0) kU/L, and the medi-an tumor size was 4.5 (1.0-27.0) cm. The expression levels of SVEP1, PKHD1, and P53 in primary liver tumors were detected by immu-nohistochemistry and assigned separate immunohistochemical scores. The areas under the ROC curves of the immunohistochemical scores of SVEP1, PKHD1, and P53 were 0.861, 0.829, and 0.716, respectively. The critical values of SVEP1, PKHD1, and P53 were 4, 4, and 1 point, respectively (P<0.001). The three-year DFS rates among the SVEP1 high-risk (expression≤4 points) and low-risk groups (expression>4 points) were 4.1% and 51.7%, respectively. Similarly, the three-year survival rates among the PKHD1 high-risk (expres-sion≤4 points) and low-risk groups (expression>4 points) were 5.3% and 51.9%, respectively. The three-year DFS rates among the P53 high-risk (expression>1 point) and the low-risk group (expression≤1 point) were 6.3% and 27.3%, respectively. The survival differenc-es between all the pairs were statistically significant (P<0.001,<0.001, and 0.003 respectively). When PKHD1 was used in combination with SVEP1, the ROC curve had an area of 0.897 (P<0.001) with a sensitivity of 76.5% and a specificity of 94.4%. Conclusions: The accu-racy of P53 data for predicting primary liver cancer recurrence is insufficient and therefore it is not recommended for use. SVEP1 and PKHD1 data achieve sufficient accuracy for predicting the recurrence of primary liver cancer. Since SVEP1 data impart a higher specifici-ty and PKHD1 data impart a higher sensitivity to the prognosis scores, the combined use of the two markers is better than being used individually.

Objective To evaluate Evans lateral lengthening calcaneal osteotomy(E-LLCOT) in the treatment oftalocalcaneal coalitions (TCCs) with forefoot abduction deformity in the teenagers.Methods From February 2014 to August 2018,11 teenaged patients (14 feet) were treated at Department of Foot and Ankle Surgery,Zhengzhou Orthopaedics Hospital for TCCs with severe forefoot abduction deformity.They were 6 males (8 feet) and 5 females (6 feet),aged from 13 to 17 years (average,15 years).Their diseases involved bilateral feet in 3 cases and unilateral foot in 8,the left foot in 7 cases and the right in 7.All patients underwent TCCs resection followed by E-LLCOT.Their talonavicular coverage angles (TCA) and talar-second metatarsal angles (T2-MT) on the anteroposterior film and talar horizontal angles (TH) and talar-first metatarsal angles (T1-MT) on the lateral film were measured preoperatively and at the last follow-up.The foot functions were evaluated preoperatively and at the last follow-up using the ankle-hindfoot scores of American Orthopaedic Foot and Ankle Society (AOFAS) and visual analogue scale (VAS).Results All the 11 patients were followed up for 12 to 24 months (average,16.5 months).The mean preoperative TCA (22.3°,from 20° to 26°) was improved to 10.5° (from 8° to 13°) at the last follow-up;the mean T-2MT was improved from preoperative 17.6° (from 16° to 20°) to 6.5° (from 5° to 11°) at the last follow-up;the mean TH on the lateral view was improved from preoperative 35° (from 25° to 40°) to 17.5° (from 16° to 21°) at the last follow-up;the mean T-1MT was improved from preoperative 15.5° (from 10° to 22°) to 3.5° (from 2° to 6°) at the last follow-up;the mean AOFAS score was improved from 56.5 (from 50 to 62) preoperatively to 90.6 (from 75 to 95) at the last follow-up;the mean VAS score was improved from 6.0 (from 5 to 7) preoperatively to 2 (from 0 to 3) at the last follow-up.Conclusion For TCCs with severe forefoot abduction deformity in the geenagers,E-LLCOT after TCCs resection can effectively correct deformity,relieve pain and achieve significant functional and radiographic improvements.

Objective:To analyze the therapeutic effect of ankle arthrodesis with reverse proximal humerus internal locking system plating plus cannulated screwing via the transfibular approach in the treatment of end-stage ankle arthrosis.Methods:From April 2014 to January 2018, 19 patients (19 ankles) with end-stage ankle osteoarthritis were treated at Department of Foot and Ankle Surgery, Zhengzhou Orthopaedics Hospital by ankle arthrodesis with reverse PHILOS plating plus cannulated screwing via the transfibular approach for multi-plane fixation. They were 9 men and 10 women, aged from 36 to 65 years (average, 46.7 years). The disorder was further diagnosed as traumatic arthritis in 9 cases, as osteoarthritis in 6, as talar necrosis in 2 and as equinovarus in 2, and affected the left ankle in 11 cases and the right in 8. All cases belonged to stage Ⅲ osteoarthritis according to the Morrey-Wiedeman imaging classification. Their American Orthopaedic Foot and Ankle Society (AOFAS) ankle-hindfoot score, visual analogue scale (VAS), joint fusion and complications were observed preoperatively and at the last follow-up.Results:All the 19 patients were followed up for 18 to 62 months (average, 31.5 months). Their AOFAS scores were improved from preoperative 46.3±2.1 to 81.6±0.9 at the last follow-up, and their VAS was reduced from preoperative 7.1±0.2 to 2.0±0.2 at the last follow-up, showing significant differences ( P<0.001). All their ankle joints obtained bony fusion after 9 to 14 weeks (mean, 11.5 weeks). No implant loosening or breakage was observed. All the patients were satisfied with their operative results at the last follow-up. Dorsal lateral foot numbness was observed in one patient who became asymptomatic after neurotrophic medication for 3 months. Lateral distal wound dehiscence happened during removal of stitches at 2 weeks after operation in another patient but healed after debridement. The last follow-up found subtalar joint degeneration in 2 cases and talonavicular joint degeneration in one but no clinical symptoms in the 3. Conclusion:Ankle arthrodesis with reverse PHILOS plating plus cannulated screwing via the transfibular approach is a fine treatment for end-stage ankle arthrosis, because it leads to reliable fixation, short fusion time, alleviated pain and improved ankle function.

Objective:To explore the outcome of sponge forceps assisted threading with Speedbridge technique for the treatment of acute closed Achilles tendon rupture.Methods:A retrospective case series study was conducted on 20 patients with acute closed Achilles tendon rupture treated in Zhengzhou Orthopedic Hospital from December 2019 to December 2021. There were 18 males and 2 females, with age range of 24-43 years [(29.5±7.6)years]. All patients were with unilateral injury, involving the left side in 13 patients and right side in 7. Examinations revealed a palpable defect in the Achilles tendon and positive Thompson test. A longitudinal incision was made at the medial edge of the ruptured tendon. Three nonabsorbable sutures were passed through the proximal stump with sponge forceps, bypassed the rupture site and fixed directly into the calcaneal bone. The disrupted tendon ends were aligned by the tendon-bundle technique using 4-0 absorbable sutures. The operation time and incision length were documented. The ankle joint range of motion (dorsiflexion/plantar flexion), American Orthopedic Foot and Ankle Society (AOFAS) ankle-hindfoot score and Achilles tendon total rupture score (ATRS) in the affected and healthy side were compared at 3, 6 and 12 months postoperatively. The wound healing and complications were observed.Results:All patients were followed up for 12-16 months [(13.2±2.5)months]. The operation time was 40-66 minutes [(52.0±10.3)minutes], with the incision length of 3-4 cm [(3.3±0.7)cm]. In the affected side at 3 and 6 months postoperatively, the ankle joint dorsiflexion [(5.6±1.5)°, (10.5±0.2)°] and plantar flexion [(28.4±3.2)°, (33.5±1.5)°] showed statistically significant difference compared with the healthy side (all P<0.05). The ankle joint dorsiflexion [(13.9±0.7)°] and plantar flexion [(38.3±4.4)°] in the affected side were not statistically different from that of the healthy side at 12 months postoperatively (all P>0.05). The AOFAS ankle-hindfoot score was (58.3±5.4)points, (84.9±7.1)points and (91.8±6.3)points at 3, 6 and 12 months postoperatively, showing a gradual rise (all P<0.05). The ATRS was (60.5±4.9)points, (85.5±9.0)points and (93.1±5.7)points at 3, 6 and 12 months postoperatively, showing a gradual rise (all P<0.05). All incisions were healed primarily. No patients had wound infection, nerve injury or re-rupture. Pain at the anchor insertion site occurred in 2 patients at 1 month after operation and relieved after active functional rehabilitation at 4 months after operation. Transient pain at the Achilles tendon insertion occurred in 1 patient at 6 months after operation, and relieved after 2 weeks of oral non-steroidal anti-inflammatory drugs treatment. Conclusion:For acute closed Achilles tendon rupture, sponge forceps assisted threading with Speedbridge technique can attain short operation time, small incision and good functional recovery, with few complications.