To reduce the risk of 3′-terminal mismatch between primers and template and increase the sensitivity of polymerase chain reaction (PCR) in the detection of variable region of DNA. Methods: A pair of special primer(WU,WD) was designed to amplify a fragment of HBV DNA P gene by PCR. Other 2 similar pairs of primer (MU1, MD1, MU2, MD2) were obtained by knocking off 1 or 2 bases at the 3′-terminal of WU and WD. (1) Special primers (WU, WD) and degeneracy primers(WU, WD, MU1, MU2, MD1, MD2) were used to amplify 27 samples respectively by PCR under the same condition. The sensitivity of each PCR was compared. (2) Using degeneracy primers, serum HBV DNA was amplified from 4 patients who were resistant to lamivudine. The PCR products were sequenced to evaluate the effect of the 3′-terminal mismatch of primers upon PCR. Results: (1) The sensitivity of special primers and degeneracy primers were 70.4%(19/27) and 85.2%(23/27) respectively (P<0.05). (2) The sequencing analysis of the PCR products suggested that the 3′-terminal mismatch of primers caused false negative in the PCR detection. Conclusion: When amplifying the variable region of DNA, the false negative result can be avoided by using 3′-terminus shifted degeneracy primers.

Objective To observe the efficacy and adverse effects of conventionally fractionated (CF) versus hypofractionated (HF) three-dimensional conformal radiotherapy (3DCRT) for hepatocellular carcinoma (HCC) with portal vein tumor thrombosis.Methods A retrospective analysis was performed on the clinical data of 65 patients with HCC suitable for 3DCRT from 2008 to 2012.These patients were randomly divided into HF group (n =32) and CF group (n =32).The HF group received 3DCRT at a total dose of 45-55 Gy (3-4 Gy/fraction,3-5 fractions/week),while the CF group at a total dose of 40-56 Gy (2 Gy/fraction,5 fractions/week).Results The follow-up rate was 100％.For the HF group and CF group,the short-term response rates were 72％ vs.55％ (P =0.034),the 1-year local control rates were 72％ vs.55％ (P=0.034),the 1-year overall survival rates were 59％ vs.45％ (P=0.042),and the numbers of individuals with grade 1-2 acute gastrointestinal reactions,deterioration of liver function,and radiation hepatitis were 14 vs.11,8 vs.8,and 1 vs.0,respectively (P =0.237).Conclusions HF 3DCRT is superior to CF 3DCRT in the treatment of HCC.

Objective: To reduce the risk of 3'-terminal mismatch between primers and template and increase the sensitivity of polymerase chain reaction (PCR) in the detection of variable region of DNA. Methods: A pair of special primer(WU, WD) was designed to amplify a fragment of HBV DNA P gene by PCR, Other 2 similar pairs of primer (MU1, MD1, MU2, MD2) were obtained by knocking off 1 or 2 bases at the 3'-terminal of WU and WD. (1) Special primers (WU, WD) and degeneracy primers(WU, WD, MU1, MU2. MD1, MD2) were used to amplify 27 samples respectively by PCR under the same condition. The sensitivity of each PCR was compared. (2) Using degeneracy primers, serum HBV DNA was amplified from 4 patients who were resistant to lamivudine. The PCR products were sequenced to evaluate the effect of the 3'-terminal mismatch of primers upon PCR. Results: (1) The sensitivity of special primers and degeneracy primers were 70. 4%(19/27) and 85. 2% (23/27) respectively (P

Objective To evaluate the functional outcomes and complications of patients with bone tumors who were treated with the excision-alcoholization-replantation (EAR) method. Methods From August 1965 to August 2003, 191 patients treated with the EAR method who had suitable follow-up were evaluated in our department, including 102 males and 89 females with an average age of 20 years (range, 10-62). On the basis of Enneking tumor staging, 136 patients were identified II B stage, 25 I B stage, and 30 invasive benign tumors and tumor-like lesion. The length of the resected part ranged from 5 cm to 26 cm, with marginal resection in 30 cases, wide resection in 116. The oncological parameters that were evaluated included survival of the patients, local recurrence, and metastasis. Complications included non-union, infection, and fracture of the grafts. Mankin evaluation system was used to assess functional outcomes. Results The mean follow-up time was 32 months (range, 1-372), 52 patients died with disease at the final follow-up. Five-year survival rates of high grade sarcoma and low grade sarcoma were 59% and 90%, respectively, with statistical significance (P=0.02). Lung metastasis were demonstrated in 52 patients (27.2%, 52/191), 51 (26.7%, 51/191) had local recurrence. Except the tumor factor, the total complication rate was 50.3%(96/191). Local complications included 33 nonunion (17.3%, 33/191), 39 fracture of the grafts (20.4%, 39/191), 39 infections of the graft (20.4%, 39/191), 15 fracture of internal fixation (7.9%, 15/191), 5 instability or semiluxation of the joint (2.6%, 5/191). The 5-year survival rate of inactivated bone was 55%. On the basis of Mankin functional evaluation, the satisfied rate was 50.3%. At last there were 141 patients (73.8%, 141/191) preserved their extremities. Conclusion Compared to other methods in the same period, EAR method is considered on the same level of safety in oncology, which superiorities are economic-applicable to patients and the well fitness of bone graft with the defects.

Objective To investigate the inhibitive effect of mesenchymal stem cells (MSCs) on the immunological rejection in rats after liver transplantation. Methods The recipients and donors were female SD rats and Wistar rats. All rats were randomly divided into 3 groups (28 rats in each group). Rats in group A were infused with normal saline; rats in group B received FK506 (0.25 mg/kg) every 2 days for 2 weeks after liver transplantation; rats in group C were injected with MSCs from male Wistar rats during liver transplantation. The pathological changes, expression of TGF-β1 and IL-10, Y chromosome location, changes of liver function and the survival of the recipients were detected on postoperative day 10. The levels of ALT and AST were analyzed by com-pletely randomised design analysis of variance, and the difference among the 3 groups were analyzed by LSD. Ridit was used to analyze the pathological grading. The survival was analyzed by Log-rank test after the Kaplan-Meier survival curve was drawn. Results The values of ALT and AST were (756±104)U/L and (635±134)U/L in group A, (197±49)U/L and (331±78) U/L in group B, (103±31)U/L and (150±38) U/L in group C, respectively. The difference in the level of ALT and AST among the 3 groups had statistical significance (F = 158, 265, P < 0.05). The liver function of rats in group B and C was better than those in group A (P < 0.01), and the liver function of rats in group C was better than those in group B. The mean values of ridit in group A, B and C were 0.8333, 0.4583 and 0.2083, respectively. The expression rates of TCG-β1 in group A, B and C were 18%±5% , 69%±20% and 85%±24% , with statistical difference among the 3 groups (F=191, P <0.01). There was a significant difference in IL-10 expression among group A (21%±5%), group B (75%±14%) and group C (91%±21%) (F=672, P<0.01). The TCG-β1 and IL-10 had strong positive expression in group B and C, and the expression of TCG-β1 and IL-10 was much stronger in group C than in group B; while the expres-sion of TCG-β1 and IL-10 was weak positive in group A. MSCs cells with Y chromosome were positively stained and were concentrated at the portal area in group C. The 50-day survival rate of rats in group A, B and C were 0, 10% and 90% , respectviely, with significant difference (χ~2=36, P < 0.01). The median survival time of rats in group C was 63 days, which was longer than that in group A and B. Conclusion Simultaneous injection of MSCs from donors during liver transplantation can inhibite the immunological rejection of recipients to the liver graft.

Objective To determine the dose-response relationship of sufentanil blunting responses to double-lumen endotracheal intubation when combined with propofol given by target-controlled infusion (TCI) in patients with pulmonary tuberculosis.Methods One hundred patients of both sexes with pulmonary tuberculosis,of American Society of Anesthesiologists physical status Ⅰ or Ⅱ,aged 24-58 yr,with body mass index ＜30 kg/m2,with Mallampati grade Ⅰ or Ⅱ,undergoing thoracic surgery under general anesthesia,were divided into Ⅰ-Ⅴ groups (n =20 each) using a random number table.Anesthesia was induced with iv sufentanil 0.35 μg/kg (group Ⅰ),0.40 μg/kg (group Ⅱ),0.45 μg/kg (group Ⅲ),0.50 μg/kg (group Ⅳ) and 0.55 μg/kg (group Ⅴ),and propofol TCI (target plasma concentration 3.5 μg/ml) and iv vecuronium 0.15 mg/kg.The patients were endotracheally intubated and mechanically ventilated.The response to double-lumen endotracheal intubation was defined as positive when mean arterial pressure increased by＞ 20％ of the baseline value and/or heart rate ＞ 90 bpm within 5 min after intubation.The median effective dose (ED50),ED95 and 95％ confidence interval (95％ CI) of sufentanil blunting the responses to double-lumen endotracheal intubation were calculated by Probit analysis.Results The ED50 (95％ CI) and ED95 (95％ CI) of sufentanil blunting the responses to intubation were 0.411 (0.370-0.441) μg/kg and 0.635 (0.556-0.888) μg/kg,respectively,when combined with propofol given by TCI.Conclusion When combined with propofol given by TCI (target plasma concentration 3.5 μg/ml),the ED50 and ED95 of sufentanil blunting the responses to double-lumen endotracheal intubation are 0.411 and 0.635 μg/kg,respectively,in patients with pulmonary tuberculosis.

OBJECTIVETo study the influence of the sequence of three-dimensional conformal radiotherapy (3DCRT) and transcatheter arterial chemoembolization (TACE) on the efficacy and toxicity of treatment in patients with hepatocellular carcinoma (HCC) with portal vein tumor thrombus (PVTT).

METHODSA total of 65 patients who were diagnosed with primary HCC with PVTT were enrolled in the study from November 2008 to March 2012 and were randomly divided into the following two groups:group A,32 patients treated with 3DCRT followed by TACE; group B,33 patients treated with TACE followed by 3DCRT.

RESULTSThe total efficacy rates of groups A and B were 68.8% and 69.7% (x² =0.232, P < 0.793). The survival rates,effective percentage of PVTT and AFP remission rates were not significantly different between group A and group B.The exacerbation rate of liver function was significantly higher for group B than for group A (P < 0.05). No serious complication was found in the follow-up period for either group.

CONCLUSIONThe combination of 3DCRT and TACE is a relatively effective local treatment for patients with primary HCC and PVTT.Compared with TACE followed by 3DCRT, 3DCRT followed by TACE may have a negative influence on liver function.

Arteries ; Carcinoma, Hepatocellular ; Chemoembolization, Therapeutic ; Combined Modality Therapy ; Humans ; Liver Neoplasms ; Portal Vein ; Radiotherapy, Conformal ; Survival Rate ; Thrombosis ; Treatment Outcome

OBJECTIVE: To carry out optical genome mapping (OGM) for a Chinese pedigree with a rare paracentric reverse insertion of chromosome 17. METHODS: A high-risk pregnant woman identified at the Prenatal Diagnosis Center of Hangzhou Women's Hospital in October 2021 and her family members were selected as the study subjects. Chromosome G banding analysis, fluorescence in situ hybridization (FISH), single nucleotide polymorphism array (SNP array) and OGM were applied to verify the balanced structural abnormality of chromosome 17 in the pedigree. RESULTS: Chromosomal karyotyping analysis and SNP array assay have identified a duplication of 17q23q25 in the fetus. Karyotyping analysis of the pregnant woman showed that the structure of chromosome 17 was abnormal, whilst SNP array has detected no abnormality. OGM revealed that the woman has carried a paracentric reverse insertion, which was confirmed by FISH. The karyotype of her husband was normal. CONCLUSION The duplication of 17q23q25 in the fetus has derived from a paracentric reverse insertion of chromosome 17 in its mother. OGM has the advantage for delineating balanced chromosome structural abnormalities.
Pregnancy ; Humans ; Female ; Pedigree ; In Situ Hybridization, Fluorescence ; Chromosomes, Human, Pair 17/genetics* ; East Asian People ; Chromosome Aberrations ; Prenatal Diagnosis ; Chromosome Mapping ; Chromosome Inversion

Objective To explore the clinical features, diagnosis and interventional management of acute deep venous thrombosis of lower extremity (LEDVT)combined with type Ⅱ heparin-induced thrombocytopenia (HITⅡ) and to improve the knowledge of this disease. Methods A retrospective review and analysis of the clinical data of the patients with acute LEDVT combined with HIT Ⅱ enrolled from January 2010 to June 2014. All of them underwent anticoagulation with low molecular weight heparin (LMWH) and the comprehensive interventional therapy at the beginning of treatment.When HIT Ⅱ was
identified, all forms of heparin and LMWH were avoided . Alternative anticoagulation was commenced with argatrobam. Adjustments in interventional therapy were taken while the short-term low-dose glucocorticoid treatment were used.The clinical manifestations, changes of PLT, 4Ts score (Warkentin 4T scoring system, 4Ts) , HIT antibody assay (ELISA) and response to therapy of the patients were analyzed and the treatment effect was observed . The efficacy of interventional therapy was evaluated according to the improvement clinical symptoms and venography. Results The incidence of acute LEDVT combined with HIT Ⅱ was 1.9%(8/416). There were 4 males and 4 females with a median age of 24 years in this study. The median time between their initiation exposure to heparin and onset of thrombocytopenia was 5 days (range,3 to 8 days). The median platelet counts prior to HIT Ⅱ was 218 × 109/L( range,122 × 109/L to 254 × 109/L ). Platelet counts decreased to the lowest level range from 20 × 109/L to 51 × 109/L(median 32 × 109/L). After alternative anticoagulation, the interval period which PLT recovered to the basic level was range from 3 to 7 days (median 3.5 days) . According to the score of 4Ts , there were 2 cases score 6 and 6 cases score 8. HIT antibody assay (ELISA) was detected in 6 patients which the results were positive. During heparin anticoagulation treatment, the LEDVT condition of all patients continued to deteriorate. Vein thrombosis extended in 7 patients. Among them, 5 patients occurred new thrombosis in the inferior vena cava and(or) at the vessel of catheter insertion. Another 2 patients complicated with pulmonary embolism. After underwent anticoagulation with argatrobam , with the increased of PLT the treatment efficacy of thrombolysis therapy was ameliorated. At the endpoint of interventional therapy, the curative effect evaluation was excellent in 3 cases, good in 3 cases and medium in 2 cases respectively. All patients were followed up for 12 to 20 months (median 15.5 months) with no evidence of recurrence .Conclusions The study showed that acute LEDVT combined with HITⅡdisplayed the following features:(1)an absolute drop in platelet count below the normal range (PLT ≤100 × 109/L) or as a relative decrease of 30% to 50% from baseline counts. (2) refractory venous thrombosis,during the interventional treatment of acute LEDVT, platelets counts should be monitored regularly in patients who receiving heparin anticoagulation. For patients with strongly suspected HIT Ⅱ, withdrawal of all forms of heparin and early introduction of alternative anticoagulant therapy can improve the effect of interventional therapy.

Objective To compare the dose, clinical efficacy and acute adverse reactions of intensity modulated radiotherapy(IMRT)and three-dimensional conformal radiotherapy(3D-CRT)combined with three-dimensional brachytherapy (3D-BT) in the treatment of concurrent radiotherapy and chemotherapy for advanced stage cervical cancer patients. Methods Data collection was performed from January 2011 to November 2015 in Chinese PLA General Hospital and Inner Mongolia Cancer Hospital.All 89 patients with advanced stage (Ⅱb-Ⅲb) cervical cancer were treated by pelvic radiotherapy and concurrent chemotherapy, 46 cases of them received IMRT and 3D-BT(IMRT group), 43 cases received 3D-CRT and 3D-BT(3D-CRT group),along with cisplatin chemotherapy.The dose accumulation of external beam radiotherapy and 3D-BT was calculated by deformable image registration to analyze clinical efficacy, acute adverse reactions and prognosis of the two groups.Results (1)Dose of radiotherapy:planning target volume(PTV)coverage of IMRT group and 3D-CRT group were respectively(95.4±4.7)% and(95.1±5.1)%, without significant differences (t=0.289, P=0.773). Compared with the patients treated with 3D-CRT, the volumn receiving at least 30 Gy (V30), V50of rectum, colon, bladder and small intestine and V20of bone marrow in the IMRT group were significantly decreased (P<0.05). Regarding the combined dose, the maximum dose (Dmax) and the minimum dose received by the most exposed 2 cm3volume of the analyzed organ(D2CC)of rectum,colon,bladder and small intestine of IMRT group were significantly lower than those of 3D-CRT group (P<0.05). (2) Short-term efficacy: the effective rate of IMRT and 3D-CRT group were respectively 93% (43/46) and 91% (39/43), with no significant differences (χ2=0.237,P=0.626). (3) Acute adverse reactions: compared with 3D-CRT, IMRT could significantly reduce grade 1-2 acute toxicity in gastrointestinal [63%(29/46) vs 84%(36/43)], genitourinary [17%(8/46) vs 37%(16/43)] and hematologic [57%(26/46) vs 79%(34/43)] system (all P<0.05). There were no significant differences of grade 3 acute adverse reactions of gastrointestinal,genitourinary and hematologic system between two groups(all P>0.05). No grade 4 acute adverse reactions were observed. (4) Prognosis: the overall survival rate at 1, 2-year of IMRT and 3D-CRT group were respectively 95.6%,89.1% and 93.1%,86.1%.The progression-free survival rateat 1, 2-year of IMRT and 3D-CRT group were 91.1%, 89.1% and 88.4%, 86.1%, respectively. There were no significant differences in overall survival rate and progression-free survival rate between two groups (P>0.05). Conclusions Compared with 3D-CRT, IMRT combined with 3D-BT has dosimetry advantages based on dose accumulation algorithms by deformable image registration. IMRT could ensure clinical efficacy and significantly reduce the incidence rate of acute toxicities.