Objective To evaluate the efficacy of hypofractionated 3DCRT for primary liver cancer(PLC) with portal vein tumor thrombosis(PVTT).Methods Between April 1999 and August 2003,34 PLC patients with PVTT received hypofractionated 3DCRT.The severity of hepatic cirrhosis was 23 in Child-Pugh gradeA and 11 gradeB.The median value of GTV was 773?cm~3(105-2097?cm~3).The radiotherapy regimen consisted of 38-63?Gy in 7-15 fractions with 4-8?Gy per fraction(median value 5?Gy),the treatment was delivered 3 times per week during every other day.Results Having response rate(CR+PR) of 76%(26/34),the overall 1-,2-,and 3-year survival rate at was 36%,19% and 13%,respectively.Conclusion Hypofractionated three-dimensional conformal radiotherapy is effective for primary liver cancer with portal vein tumor thrombosis.

Objective:To explore the mechanism of dendritic epidermal T lymphocytes (DETCs) in promoting healing of full-thickness skin defect wound on mice by regulating the proliferation and differentiation of epidermal stem cells (ESCs) in mice.Methods:(1) Ten 8-week-old wild type (WT) male C57BL/6 mice (the same sex and kind below) were sacrificed to collect the skin of back for extracting DETCs to culture. Five WT and five 8-week-old T cell receptor (TCR) δ -/ - mice were selected and enrolled in WT control group and TCR δ -/ - control group, respectively. A full-thickness skin defect wound with diameter of 6 mm was made on both sides of spinal line on the back of mice without any treatment after injury. Another fifteen 8-week-old TCR δ -/ - mice were selected and divided into phosphate buffer solution (PBS), DETC, and insulin-like growth factor-Ⅰ(IGF-Ⅰ) groups according to the random number table (the same grouping method below), with 5 mice in each group, and the same full-thickness skin defect wound was made on each mouse. Immediately after injury, mice in PBS, DETC, and IGF-Ⅰ groups were injected subcutaneously around each wound with 10 μL sterile PBS , DETCs (cell concentration of 1×10 6/mL), and 5 mg/mL recombinant mice IGF-Ⅰ, respectively. The percentage of the residual wound area was calculated on post injury day (PID) 2, 4, 6, and 8. (2) Three 8-week-old WT mice were enrolled in WT control group and nine 8-week-old TCR δ -/ - mice were divided into TCR δ -/ - control group, PBS group, and DETC group, with 3 mice in each group. The full-thickness skin defect wound was made as in experiment (1) . On PID 3, the protein expression of IGF-Ⅰ in the epidermis tissue of wound margin was detected by chemiluminescence imaging analyzer. (3) Three 8-week-old WT mice were enrolled in WT control group and six 8-week-old TCR δ -/ - mice were divided into PBS and DETC groups, with 3 mice in each group, and the full-thickness skin defect wound was made as in experiment (1). On PID3, DETCs were extracted from the wound margin epidermis tissue to detect the percentage of DETCs expressing IGF-Ⅰ by flow cytometer. (4) The mice were taken as in experiment (2) and divided into WT control, PBS, DETC, and IGF-Ⅰ groups. A straight full-thickness skin defect incision with length of 3 cm was made in the direction of one inner ear. Mice in WT control group didn′t have any other treatment after injury, and immediately after injury, mice in PBS, DETC, and IGF-Ⅰ groups were injected subcutaneously around each wound with 10 μL sterile PBS, DETCs (cell concentration of 1×10 6/mL), and 5 mg/mL recombinant mice IGF-Ⅰ, respectively. On PID 12, epidermis tissue of wound margin was collected, and immunofluorescence staining was performed to observe the number of keratin 15 positive cells. (5) The same mice were collected, grouped, and treated as in experiment (4). On PID12, the epidermis tissue of wound margin was collected and immunofluorescence staining was performed to observe the number of keratin 10 positive cells. (6) Twenty 3-day-old WT mice (the same below) were sacrificed to collect the whole skin, which was used to extract ESCs, with 5 mice detecting one index. The ESCs were divided into DETC co-culture group and control group, which were added with 1 mL DETCs (cell concentration of 1.25×10 6/mL) and DETC medium, respectively. The percentage of 5-ethynyl-2′-deoxyuridine (EdU) positive cell on culture day (CD) 3, the percentages of CD49f + CD71 - and keratin 14 positive cells on CD 5, and the percentage of keratin 10 positive cell on CD 10 in 2 groups were detected by flow cytometer. (7) Twenty mice were taken to extract ESCs, with 5 mice detecting one index. The ESCs were divided into control group and IGF-Ⅰ group, which were added with 1 mL sterile PBS and 10 ng/mL recombinant mice IGF-Ⅰ, respectively. The percentages of EdU positive cell, CD49f + CD71 - cell, keratin10 positive cell, and keratin 14 positive cell were detected as in experiment (6). The sample in each group of experiments (6) and (7) was three. Data were statistically analyzed with analysis of variance for repeated measurement, one-way analysis of variance, and t test. Results:(1) On PID 4, 6, and 8, the percentage of residual wound area in TCR δ -/ - control group was significantly higher than that in WT control group ( t=2.78, 3.39, 3.66, P<0.05 or P<0.01). The percentage of residual wound area in DETC group and IGF-Ⅰgroup on PID 4, 6, and 8 was apparently lower than that in PBS group ( t=2.61, 3.21, 3.88, 2.84, 2.91, 2.49, P<0.05 or P<0.01). (2) On PID 3, the protein expression of IGF-Ⅰ in the epidermis tissue of wound margin of mice in TCR δ -/ - control group was significantly lower than that in WT control group ( t=17.34, P<0.01). The protein expression of IGF-Ⅰ in the epidermis tissue of wound margin of mice in DETC group was significantly higher than that in PBS group ( t=11.71, P<0.01). (3) On PID 3, the percentage of DETCs expressing IGF-Ⅰ in the epidermis tissue of wound margin of mice in PBS group was significantly lower than that in WT control group and DETC group ( t=24.95, 27.23, P<0.01). (4) On PID 12, the number of keratin 15 positive cells in the epidermis tissue of wound margin of mice in PBS group was significantly lower than that in WT control group, DETC group, and IGF-Ⅰ group ( t=17.97, 11.95, 7.63, P<0.01). (5) The number of keratin 10 positive cells in the epidermis tissue of wound margin of mice in PBS group was significantly higher than that in WT control group, DETC group, and IGF-Ⅰ group ( t=11.59, 9.51, 3.48, P<0.05 or P<0.01). (6) The percentages of EdU positive cells on CD 3, CD49f + CD71 - cells on CD 5, and keratin 14 positive cells on CD 5 in DETC co-culture group were respectively (43.5±0.6)%, (66.5±0.5)%, (69.3±1.7)%, apparently higher than (32.3±1.3)%, (56.4±0.3)%, (54.9±1.3)% in control group ( t=7.97, 17.10, 6.66, P<0.01). The percentage of keratin 10 positive cells on CD 10 in DETC co-culture group was (55.7±0.7)%, significantly lower than (67.1±1.2)% in control group ( t=8.34, P<0.01). (7) The percentages of EdU positive cells on CD 3, CD49f + CD71 - cells on CD 5, and keratin 14 positive cells on CD 5 in IGF-Ⅰ group were respectively (42.1±0.9)%, (81.1±1.3)%, (66.8±1.0)%, apparently higher than (32.4±0.7)%, (74.9±0.7)%, (52.0±1.9)% in control group ( t=8.39, 4.24, 7.25, P<0.05 or P<0.01). The percentage of keratin 10 positive cells on CD 10 in IGF-Ⅰ group was (53.5±1.1)% , significantly lower than (58.2±0.3)% in control group ( t=3.99, P<0.05). Conclusions:DETCs can promote the proliferation and anti-apoptotic potential of ESCs and inhibit their differentiation into end-stage by secreting IGF-Ⅰ, thus promoting wound healing of full-thickness skin defects in mice.

Objective To establish a simultaneous quantitative determination method for six lactones (santamarine, costunolide, dehydrocostuslactone, alantolatone, isoalantolactone and 11,13-dihydroi-soalantolantone ) from nutmeg-5, a Mongolian medicine compound. Methods HPLC analysis was performed by an Elite C18 column(4. 6 mm × 250 mm,5 μm) at temperature 30℃ with gradient elution (acetonitrile and aqueous solution) at a flow rate of 0. 8 mL/min 204 nm. Results The six lactones were separated completedly by using the established HPLC method, and the regression equation of each lactone showed good linear correlation between concentration and peak area with the correlation coefficient more than 0. 999. Each average recovery rate with relative standard deviation (RSD) of six lactones was listed as the following:santamarine 102 . 54% ( RSD 1 . 02%) , costunolide 103. 35% ( RSD 0 . 82%) , dehydrocostuslactone 100 . 38% ( RSD 1 . 75%) , alantolatone 97 . 81% ( RSD 1 . 59%) , isoalantolactone 96. 65% (RSD 1. 98%), and 11,13-dihydroisoalantolantone 99. 64% (RSD 1. 68%). Conclusion The HPLC method was validated and found to be specific, linear, precise, and accurate. This chromato-graphic system can be readily used on a routine way for determination of lactones from nutmeg-5 .

Objective:To investigate the feasibility and safety of a treatment system for stentless discharge after flexible ureteroscopic lithotripsy assisted by flexible negative pressure sheath.Methods:The clinical data of 72 patients with upper urinary calculi admitted to the Second Hospital of Tianjin Medical University from November 2022 to February 2023 were retrospectively analyzed. All patients achieved stentless discharge after flexible ureteroscopic lithotripsy assisted by flexible negative pressure sheath. There were 50 males and 22 females. The average age was (54.7±12.1) years. Preoperative urine culture was positive in 14 cases, negative in 3 cases (4.2%)with nitrite positive, and 11 cases were negative for urine culture and nitrites but positive of white blood cells (+ + + ). There were 29 cases of renal calculi, 33 cases of upper ureteral calculi, and 10 cases of upper ureteral calculi combined with renal calculi.The mean stone diameter was 17.0(14.0, 24.0)mm. CT value was (1 049.3±258.6)HU. Twenty-four patients carried ureteral stents before operation. A total of 42 cases used ureteral sheaths with diameters of F11/13, and 30 cases used sheaths with diameters of F12/14.During the operation, an infusion pump was used to provide sufficient irrigation pressure. The negative pressure suction was attached to the distal end of the sheath. The flexible head of the sheath was guided to the target renal calyx, to completely aspirate stone fragments. Stone baskets was used in 11 cases during the procedure. The level of ureteral injury was assessed according to the Traxer grading system at the end of the operation. A ureteral stent with extraction string was retained.On the first day postoperatively, CT scanning was performed to evaluate the residual stone fragments. Patients were discharged on postoperative day 2-3 after the removal of the ureteral stent and catheter. Follow-up was conducted for 30 days postoperatively, during which the Ureteral Stent Symptom Questionnaire (USSQ) was used to assess voiding symptoms and pain. Painkiller usage and emergency revisit situations were recorded. CT scans were performed to evaluate the stone-free rate on postoperative day 30.Results:The average operation time was 30.0 (20.0, 44.5) minutes. A total of 70 cases had no ureteral injuries, and 2 cases had Grade 1 ureteral injuries (minor mucosal damage). Three cases developed fever within 72 hours postoperatively, with no cases of septic shock or fever after stent removal. Eight patients reported waist and abdominal discomfort after discharge and took oral pain medication. Among them, one patient returned to the emergency department for pain treatment. Five patients reported moderate or severe genitourinary symptoms (including voiding frequency, nocturia, urgency/incontinence, dysuria, hematuria, and incomplete emptying) based on subjective evaluation. All patients could work and recovered a normal daily life after discharge and there was no readmission or additional surgical procedures. There were 61 patients achieved immediate stone-free status on the first day after surgery, and 66 patients achieved stone-free status during follow-up at 30 days postoperatively.Conclusions:Stentless discharge after flexible ureteroscopic lithotripsy assisted by flexible negative pressure sheath is safe and feasible.

Objective: To explore effects of dendritic epidermal T cells (DETCs) and Vγ4 T lymphocytes on proliferation and differentiation of mice epidermal cells and the effects in wound healing of mice. Methods: (1) Six C57BL/6 male mice aged 8 weeks were collected and divided into control group and wound group according to random number table (the same grouping method below), with 3 mice in each group. A 4 cm long straight excision with full-thickness skin defect was cut on back of each mouse in wound group, while mice in control group received no treatment. On post injury day (PID) 3, mice in 2 groups were sacrificed, and skin within 5 mm from the wound margin on back of mice in wound group and normal skin on corresponding part of mice in control group were collected to make single cell suspensions. The percentage of Vγ4 T lymphocyte expressing interleukin-17A (IL-17A) and percentage of DETCs expressing insulin-like growth factor Ⅰ (IGF-Ⅰ) were detected by flow cytometer. (2) Ten C57BL/6 male mice aged 8 weeks were collected and divided into control group and Vγ4 T lymphocyte depletion group with 5 mice in each group. Mice in Vγ4 T lymphocyte depletion group were injected with 200 g Vγ4 T lymphocyte monoclonal neutralizing antibody of Armenian hamster anti-mouse intraperitoneally, and mice in control group were injected with the same amount of Armenian hamster Ig intraperitoneally. One hole with full-thickness skin defect was made on each side of spine of back of each mice. The wound healing was observed on PID 1-8, and percentage of remaining wound area was calculated. (3) Six C57BL/6 male mice aged 8 weeks were grouped and treated in the same way as in experiment (2), with 3 mice in each group. On PID 3, expressions of IL-17A and IGF-Ⅰ in epidermis on margin of wound were detected with Western blotting. (4) Thirty C57BL/6 male mice aged 3 days were sacrificed, and epidermal cells were extracted. The keratin 14 positive cell rate was examined by flow cytometer (the same detecting method below). (5) Another batch of mouse epidermal cells were collected and divided into control group, IGF-Ⅰ group, and IL-17A group, with 3 wells in each group (the same well number below). Cells in IGF-Ⅰ group and IL-17A group were added with 1 mL recombinant mouse IGF-Ⅰ and IL-17A with final mass concentration of 100 ng/mL respectively, while cells in control group were added with the same amount of sterile phosphate buffered saline (PBS). On post culture day (PCD) 5, keratin 14 negative cell rate was examined. Another batch of mouse epidermal cells were collected, grouped, and treated in the same way as aforementioned experiment, and keratin 10 positive cell rate was examined on PCD 10. (6) Another batch of mouse epidermal cells were collected and added with 4 mmol/L 5(6)-carboxyfluorescein diacetate N-succinimidyl ester (CFSE) solution, and divided into control 0 d group, control 7 d group, IGF-Ⅰ group, and IL-17A group. Cells in IGF-Ⅰ group and IL-17A group were treated in the same way as the corresponding groups in experiment (5), and cells in control 0 d group and control 7 d group were treated in the same way as the control group in experiment (5). The CFSE fluorescence peaks were examined on PCD 0 of control 0 d group and PCD 7 of the other 3 groups. (7) Another batch of mouse epidermal cells were collected and divided into control group and IGF-Ⅰ group. Cells in IGF-Ⅰ group were added with 1 mL recombinant mouse IGF-Ⅰ with final mass concentration of 100 ng/mL, and cells in control group were added with the same amount of sterile PBS. On PCD 5, cells were underwent keratin 14 staining and CFSE staining as aforementioned, and keratin 14 negative cell rate of CFSE positive cells was examined. Another batch of mouse epidermal cells were collected and divided into control group and IL-17A group. Cells in IL-17A group were added with 1 mL recombinant mouse IL-17A with final mass concentration of 100 ng/mL, and cells in control group were added with the same amount of sterile PBS. On PCD 5, keratin 14 negative cell rate of CFSE positive cells was examined. Data were processed with one-way analysis of variance and t test. Results: (1) On PID 3, percentage of DETC expressing IGF-Ⅰ in normal epidermis of control group was (9.9±0.8)%, significantly lower than (19.0±0.6)% of epidermis around margin of wound group (t=8.70, P<0.01); percentage of Vγ4 T lymphocyte expressing IL-17A in normal epidermis of control group was (0.123±0.024)%, significantly lower than (8.967±0.406)% of epidermis around margin of wound group (t=21.77, P<0.01). (2) On PID 1-4, there was obvious inflammatory reaction around wounds of mice in control group, and on PID 5-8, the wound area was still large. On PID 1-4, there was slight inflammatory reaction around wounds of mice in Vγ4 T lymphocyte depletion group, and on PID 5-8, the wound area was significantly reduced. On PID 3-7, percentages of residual wound area in Vγ4 T lymphocyte depletion group were significantly lower than those in control group (t=5.92, 5.74, 7.17, 5.38, 5.57, P<0.01), while percentages of residual wound area in two groups on PID 1, 2, 6 were similar (t=1.46, 3.17, 3.10, P>0.05). (3) On PID 3, compared with those in control group, expression of IL-17A and IGF-Ⅰ in epidermis around wound margin of mice in Vγ4 T lymphocyte depletion group was markedly decreased and increased respectively (t=8.47, 19.24, P<0.01). (4) The keratin 14 positive cell rate of mouse epidermal cells was 94.7%. (5) On PCD 5, the keratin 14 negative cell rate of mice in control group was markedly higher than that of IGF-Ⅰ group, while significantly lower than that of IL-17A group (t=7.25, 5.64, P<0.01). On PCD 10, the keratin 10 positive cell rate of mice in control group was significantly higher than that of IGF-Ⅰ group, while significantly lower than that of IL-17A group (t=3.99, 10.82, P<0.05 or P<0.01). (6) Compared with that of control 0 d group, CFSE fluorescence peaks of mouse epidermal cells in control 7 d group, IGF-Ⅰ group, and IL-17A group on PCD 7 shifted to the left. Compared with that of control 7 d group, CFSE fluorescence peaks of mouse epidermal cells in IGF-Ⅰ group and IL-17A group on PCD 7 shifted to the left. (7) On PCD 5, keratin 14 negative cell rate of CFSE positive cells of mice in control group was significantly higher than that in IGF-Ⅰ group (t=9.91, P<0.01), and keratin 14 negative cell rate of CFSE positive cells of mice in control group was markedly lower than that in IL-17A group (t=6.49, P<0.01). Conclusions In the process of wound healing, IGF-Ⅰ secreted by DETC can promote the proliferation of mouse keratin 14 positive epidermal cells and inhibit their terminal differentiation, while IL-17A secreted by Vγ4 T lymphocyte can promote the proliferation and terminal differentiation of mouse keratin 14 positive epidermal cells, thus both IGF-Ⅰ and IL-17A can affect wound healing.

Objective: To explore the effects of dendritic epidermal T cells (DETC) on proliferation and apoptosis of epidermal cells in wound margin of mice and its effects on wound healing. Methods: Twenty-eight healthy specific pathogen free (SPF) C57BL/6 wild-type (WT) male mice aged 8-12 weeks and 60 SPF T lymphocyte receptor δ-knockout (TCR δ^-/-) male mice aged 8-12 weeks were selected to conduct the following experiments. (1) Eight WT mice were selected to isolate epidermal cells and primarily culture DETC according to the random number table. Morphological observation and purity identification of DETC by flow cytometer were detected immediately after culture and on culture day (CD) 15 and 30, respectively. (2) According to the random number table, 5 WT mice and 5 TCR δ^-/- mice were selected and enrolled into WT control group and TCR δ^-/- group. Round full-thickness skin defect with diameter of 6 mm was made on the back of each mouse. The wound healing condition was observed immediately after injury and on post injury day (PID) 2, 4, 6, 8, 10, and the percentage of residual wound area was calculated. (3) Mice were selected to group and reproduce model of full-thickness skin defect as in experiment (2). On PID 3, the tissue of wound margin was collected for hematoxylin eosin staining, and the length of new epithelium was measured. (4) Mice were selected to group and reproduce model of full-thickness skin defect as in experiment (2). On PID 3, epidermal tissue of wound margin was collected to determine expression of proliferating cell nuclear antigen (PCNA) using Western blotting for evaluation of proliferation of epidermal cell. (5) Mice were selected to group and reproduce model of full-thickness skin defect as in experiment (2). On PID 3, epidermal tissue of wound margin was selected and digested into single-cell suspension, and apoptosis of cells was detected by flow cytometer. (6) Forty TCR δ^-/- mice were selected to carry out the same treatment as in experiments (2)-(5). According to the random number table, these mice were enrolled into TCR δ^-/- control group and TCR δ^-/-+ DETC group, with 5 mice in each group for each experiment. Round full-thickness skin defect was made on the back of each mouse. DETC in the number of 1×10⁵ (dissolution in 100 μL phosphate with buffer purity above 90%) were injected through multiple points of wound margin of mice in TCR δ^-/-+ DETC group immediately after injury, and equal volume of phosphate buffer was injected into mice of TCR δ^-/- control group with the same method as above. Data were processed with one-way analysis of variance for repeated measurement, t test, and Bonferroni correction. Results: (1) Along with the culture time elapse, the number of dendritic structures of DETC increased gradually. The percentage of T lymphocytes was 4.67% and 94.1% of these T lymphocytes were DETC. The purity of DETC on CD 15 was 18.50% and the purity of DETC on CD 30 was 98.70%. (2) Immediately after injury, the wound healing condition of mice in WT control group and TCR δ^-/- group was similar. The wound healing speed of mice in TCR δ^-/- group was slower than that in WT control group on PID 2-10. The percentages of residual wound area of mice in TCR δ^-/- group on PID 2, 4, 6, 8, and 10 were increased significantly compared with those in WT control group (t=3.492, 4.425, 4.170, 4.780, 7.318, P<0.01). (3) The length of new epithelium of mice in TCR δ^-/- group on PID 3 was (359 ± 15) μm, which was obviously shorter than that in WT control group [(462±26) μm, t=3.462, P<0.01]. (4) Immediately after injury, wound condition of mice in TCR δ^-/-+ DETC group and TCR δ^-/- control group was similar. Compared with TCR δ^-/-+ DETC group, the wound healing speed of mice in TCR δ^-/- control group were obviously slower on PID 2-10. The percentages of residual wound area of mice in TCR δ^-/-+ DETC group on PID 2, 4, 6, 8, and 10 were decreased significantly compared with those in TCR δ^-/- control group (t=2.308, 3.725, 2.698, 3.707, 6.093, P<0.05 or P<0.01). (5) On PID 3, the length of new epithelium of mice in TCR δ^-/-+ DETC group was (465±31) μm, which was obviously longer than that in TCR δ^-/- control group [(375±21) μm, t=2.390, P<0.05]. (6) On PID 3, PCNA expression of epidermal cell in wound margin of mice in TCR δ^-/- group was 1.25±0.04, which was obviously lower than that in WT control group (2.01±0.09, t=7.415, P<0.01). (7) On PID 3, PCNA expression of epidermal cell in wound margin of mice in TCR δ^-/-+ DETC group was 1.62±0.08, which was significantly higher than that in TCR δ^-/- control group (1.05±0.14, t=3.561, P<0.05). (8) On PID 3, apoptosis rate of epidermal cell in wound margin of mice in TCR δ^-/- group was (16.1±1.4)%, which was higher than that in WT control group [(8.1±0.6)%, t=5.363, P<0.01]. (9) On PID 3, apoptosis rate of epidermal cell in wound margin of mice in TCR δ^-/-+ DETC group was (11.4±1.0)%, which was obviously lower than that in TCR δ^-/- control group [(15.4±1.4)%, t=2.377, P<0.05]. Conclusions DETC participates in the process of wound healing though promoting the proliferation of epidermal cells in wound margin and inhibit the apoptosis of these cells.