Background Trans-corneally subretinal injection in rodent model is a useful method for genetic therapy,stem cell transplantation and the study on the ophthalmic research.Standarized operation process is critical for the successful treatment.However,there is no literature to report the detailed procedure and the influence of this technique on morphology and function of retina.Objective This sudy was to introduce a method of trans-corneally subretinal injection and evaluate its influence on the morphology and function of retina.Methods Trans-corneallly subretinal injection was performed on the left eyes of 2-month-old SPF C57BL/6J mice after dilation of pupils.A 301/2G disposable needle was used to puncture the cornea within the pupil area near limbus and avoid touching the lens and irises under eye surgery microscope.Then,a 33G blunt needle was used to insert into the vitreous and toward subretinal space via corneal puncture.Normal saline with 0.1％ fluorescein sodium of 1 μl was slowly injected into the space,and 2.5％ hydroxypropyl methylcellulose was dropped on ocular surface for the observation of the fundus clearly.According to the percentage of the retina filled with subretinally injected solution,the experimental eyes were divided into 80％-100％ area group,50％-70％ area group after injection,and the mice in the pseudo-injected group,in which injection procedure stopped just before the solution was pushed in to the subretinal space did not inject any solution after punctured.The right uninjected eyes of the mice served as normal control group.Four eyes were selected for each group.The structural changes were evaluated by optical coherance tomograpby (OCT) 1 day,2 days,3 days and 5 weeks after injection,and retinal function was assessed by the recored of electroretinography (ERG) 5 weeks after injection.The retinal sepcimens were prepared to examin the morphological changes by hematoxylin and esosin staning.The use of care followd the Regulations for the Administration of Affair Concerning Experimental Animals of Zhejiang Province.Results About 70％ of the injected eyes showed that retinal blebs filled with injected green fluorescein solution occupied 50％ or more retinal area with minimal damages.The focal detachment between neurosensory retinal layer and retinal pigment epithelium (RPE) was exhibited 1 day postinjection,and almost all the retinas retached 2 days after injection.In the fifth week after injection,the amplitudes of ERG b wave were (386.25±37.88),(357.50±41.03),(324.25±53.45) and (410.50±14.88) μV in the sham operation group,50％-70％ area group,80％-100％ area group and normal control group,respectively,showing a significant difference among the 4 groups (F=3.574,P=0.047),and the amplitudes of b wave in the normal control group were higher than those in the 80％-100％ area group (all at P ＜ 0.05).The detachment between retinal neuroepithelium layer and RPE layer,cell proliferation and transposition in the outer nuclear layer were dispalyed under the light microscope in the sham operation group,50％-70％ area group and 80％-100％ area group,and the disordered outer segment of photoreceptors at the injecting area was seen in the 50％-70％ area and 80％-100％ area groups at five weeks after injection.However,retinal sructure and morphology were normal at the non-injection area.Conclusions Trans-corneally subretinal injection is an effective and safe way for subretinal injection.

Stargardt disease (STGD) is an inherited disorder of retinal pigment epithelium.Three genes have been found to be implicated in STGD including Abca4 (adenosine triphosphate-binding cassette,sub-family A,member 4),Elovl4 (elongation of very long chain fatty acids protein 4) and Prom1 (prominin-1).Target genes can be delivered to the retina by various methods such as lentivirus (LV) vectors,adeno-associated virus (AAV) vectors and non-viral nano-particles.The Abca4-/-,Elovl4-/-and Prom1-/ mice model are used to study the pathogenesis mechanism and treatment of STGD.Retinal function improved significantly upon gene therapy in these models.Based on these works using animal model,phase Ⅰ /Ⅱ a clinical trial of Abca4-associated STGD gene therapy are underway.As a LV vector,equine infectious anemia virus (EIAV) is used to carry the Abca4 gene.These studies will evaluate three dose levels of the EIAV vector for safety,tolerability and biological activity.Moreover,some preclinical attempts to deliver Abca4 via AAV have been made using a modified AAV vectors because of the large size of the ABCA4 cDNA.The good responses in animal models render STGD a very attractive object for human gene therapy after the successful of the phase Ⅰ /Ⅱ clinical trials of Leber's congenital amaurosis.

Objective To explore the relationship between schizotypal personality proneness and intelligence .Methods 1905 subjects were tested by Chinese Soldier Personality Questionnaire(CSPQ).According to the results of CSPQ,the subjects were divided into high-risk subjects and normal ones.The Words Reasoning Test, the Arithmetic Reasoning Test and the Assembling Test were used to evaluate the intelligence,and the scores of these tests were compared between the two groups.Correlation coefficient between schizotypal personality proneness and intelligence was calculated.Results The scores of the Words Reasoning Test,the Arithmetic Reasoning Test and the Assembling Test in high-risk subjects were significantly lower than those in normal ones((87.83±18.42)VS(101.37±13.48),P<0.01;(91.74±14.26)vs(101.65±14.62),P<0.01;(87.70±18.82)VS (101.73±14.26),P<0.01).Schizotypal personality proneness was significantly correlated with the scores of intelligence(r=-0.39,P<0.01).Conclusion Lower intelligence level is found in the group with schizotypal personality proneness.Intelligence is correlated with schizotypal personality proneness.

Objective To evaluate the relationship and bias of the Stago-CT and CA1500 automatic coagulation analyzer.Meth-ods The relationship and bias of PT,APTT,INR,FIB,TT,DD examined by the Stago-CT and CA1500 automatic coagulation ana-lyzer by using NCCLS EP9-A2.Results For the six items(PT,APTT,INR,FIB,TT,DD)the r2 were 0.996 9,0.969 1,0.967 7, 0.955 8,0.972 6,0.949 6,respectively,and the bias were 2.9,0.88,5.22,1.16,3.48,20.3.Conclusion The five items (PT, APTT,INR,FIB,TT)at a good relationship(r2 >0.95)by the Stago-CT and CA1500 automatic coagulation analyzer except for the DD(r2 =0.949 6);The bias of the five items(PT,APTT,INR,FIB,TT)were within in the United States of demanding that a third of the clinical laboratory of CLIA 88′bias,except for the DD.

Objective To investigate the efficacy and safety ofQingre Huoxue Mixture in the treatment of blood heat type psoriasis vulgaris.Methods The patients were divided into treatment group and control group according to random number table method, with 52 cases in treatment group and 50 cases in the control group, 10 cases failed to complete the trial. Treatment group was givenQingre Huoxue Mixture, and the control group received oral administration of compoundQingdaiCapsule. The treatment lasted for 8 weeks. PASI score, TCM syndrome integrals, DLQI score, serum IFN-γ and IL-10 were detected, and hematuria routine and liver and kidney function were under safety testing before and after treatment.Results After treatment, PASI score, DLQI score and TCM syndrome integrals in the treatment group decreased significantly (P<0.001); the expression of serum IFN-γ decreased significantly (P<0.001); the expression of serum IL-10 increased significantly (P<0.001), with the total effective rate of 100% (52/52). After the treatment, TCM syndrome integrals in the both groups decreased significantly (P<0.001). After the treatment, the rash burning and itching scores in the treatment group were lower than the control group, with statistical significance (P<0.001). In terms of safety, the treatment group had no abnormal indicators.ConclusionQingre Huoxue Mixture has obvious curative effect, which can reduce the patients with psoriasis, improve the life quality of patients, and improve the clinical symptoms of rash burning and itching, reduce serum IFN-γ, and increase serum IL-10.

BACKGROUND: Recent studies have shown that induced pluripotent stem cells (iPSCs) could be differentiated into mesenchymal stem cells (MSCs) with notable advantages over iPSCs per se. In order to promote the application of iPSC-MSCs for osteoregenerative medicine, the present study aimed to assess the ability of murine iPSC-MSCs to differentiate into osteoblast phenotype. METHODS: Osteogenic differentiation medium, blending mouse osteoblast-conditioned medium (CM) with basic medium (BM) at ratio 3:7, 5:5 and 7:3, were administered to iPSC-MSCs, respectively. After 14 days, differentiation was evaluated by lineage-specific morphology, histological stain, quantitative reverse transcription-polymerase chain reaction and immunostaining. RESULTS: The osteogenesis-related genes, alp, runx2, col1 and ocn expressions suggest that culture medium consisting of CM:BM at the ratio of 3:7 enhanced the osteogenic differentiation more than other concentrations that were tested. In addition, the alkaline phosphatase activity and osteogenic marker Runx2 expression demonstrate that the combination of CM and BM significantly enhanced the osteogenic differentiation of iPSC-MSCs. CONCLUSION: In summary, this study has shown that osteoblast-derived CM can dramatically enhance osteogenic differentiation of iPSC-MSCs toward osteoblasts. Results from this work will contribute to optimize the osteogenic induction conditions of iPSC-MSCs and will assist in the potential application of iPSC-MSCs for bone tissue engineering.
Alkaline Phosphatase ; Animals ; Bone and Bones ; Culture Media, Conditioned ; Induced Pluripotent Stem Cells ; Mesenchymal Stromal Cells ; Mice ; Osteoblasts ; Phenotype

We report the induced labor of conjoined twins in the second trimester in a woman with a history of two previous cesarean sections, the last one of which was performed in 2017. This 25-year-old patient was found to have thoracolumbar conjoined fetuses with one heart and polyhydramnios through the routine ultrasound examination at 19 +5 gestational weeks and was admitted at 20 +1 gestational weeks. After a full assessment of the fetal and maternal condition through multidisciplinary consultation, it was determined to attempt a vaginal delivery as no absolute contraindication for induction of labor. The patient was given 300 mg mifepristone orally plus an amniotic cavity injection of 100 mg ethacridine lactate. Regular contractions occurred 28 hours after medication. The patient delivered a pair of dead female conjoined twins at 20 +6 gestational weeks following successful induction of labor, with an assisted vaginal breech delivery. There was no soft tissue damage in the birth canal, and the estimated blood loss was 150 ml. Pathological examination and autopsy showed thoracolumbar conjoined deformity twins with a common heart and liver. Adequate prenatal evaluation, a detailed understanding of the indications for induction of labor and vaginal delivery, closed monitoring during labor, and preparation for emergency cesarean section, are essential safety measures for induced labor of conjoined twins in women with a scarred uterus in the second trimester.

We report the diagnosis and treatment of a rare case of epidural analgesia failure followed by postpartum subdural hematoma. The patient underwent vaginal delivery under epidural analgesia at 32 +6 gestational weeks due to threatened premature labor, during which an unexpected dural rupture occurred. She gave no history of headache and there was no obvious abnormality during the pregnancy. However, on postpartum day 4, the patient complained of headache that could not be relieved when supine, but without any other neurological symptoms. A prompt cranial CT examination showed a left frontotemporal subdural hematoma. After conservative management with intravenous drip of mannitol, re-examination of cranial CT showed that the left frontotemporal subdural hematoma was mostly absorbed and the patient was discharged on postpartum day 18. The patient was healthy during follow up. Intracranial subdural hematoma after dural puncture is a rare and serious complication that requires early recognition and treatment.

Objective:To examine the preliminary clinical efficacy of Chinese magnetic sphincter augmentation (MSA) in the treatment of gastroesophageal reflux disease (GERD).Methods:According to the enrollment criteria for the MSA developed by ShengJieKang Co. and Shanghai Chest Hospital (SS-MSA) clinical trial, a total of 19 GERD patients were treated with SS-MSA from August 2018 to January 2020 at Department of Thoracic Surgery, Shanghai Chest Hospital, Shanghai Jiao Tong University. The majority of registered cases were male patients with age of (32.2±7.3) years (range: 22 to 50 years), height of (170.7±6.2) cm (range: 160 to 179 cm) and weight of (65.2±10.3) kg (range: 47.5 to 90.0 kg). SS-MSA was implanted via laparoscopy. The major evaluation indexs of postoperative efficacy were the total time of acid exposure within 24 hours and the total number of reflux. Secondary efficacy indicators included: (1) evaluation of the average daily dose of proton pump inhibitor medications; (2) the score of GERD health related quality of life questionnaire (GERD-Q) before and after MSA implantation. Paired design t-test was used to evaluate the efficacy of the SS-MSA. Results:A total of 19 patients underwent SS-MSA surgery successfully. The history of the GERD were 19 (54) months ( M( QR)). The operation time was 63 (22) minutes and the in-hospital stay was 3 (2) days. No obvious surgical complications occurred. Postoperative adverse events included 14 cases with mild to moderate dysphagia exited after surgery, gradually eased within 1 to 3 months, 1 case with the removal of the device after 1 month of severe swallowing difficulties, 1 case of diarrhea. No corrosion, perforation, displacement occurred. The GERD-Q score (11.0(4.5) vs. 6(1.0), t=4.274, P=0.013), 24-hour acid exposure time (6.2(4.8)% vs. 0.1(0.9)%, t=5.814, P=0.004), and Demeester score (23.72(16.20) vs. 0.96(3.10), t=6.678, P=0.003) were significantly decreased 1 year after surgery( n=5). Proton pump inhibitor reuse rates were 6/18, 5/15, 3/10, and 1/5 in 1, 3, 6 and 12 months after the operation, respectively. Conclusions:SS-MSA implantation is feasible and safe with short hospital stay and rare perioperative complications. The preliminary results is good after 1 year follow-up. It could be expected to be an ideal substitutive for future GERD treatment.

Objectives:To examine the prognosis factors of recurrence of esophageal carcinoma within 6 months after neoadjuvant therapy followd by surgery.Methods:The clinical data of 187 patients with esophageal squamous cell carcinoma who underwent neoadjuvant therapy followed by curative esophagectomy between January 2018 and April 2020 at Department of Thoracic Surgery, Shanghai Chest Hospital were analyzed retrospectively. There were 160 males and 27 females, aging (63.0±7.1) years (range:43 to 76 years). The t test, χ 2 test and rank-sum test were used for univariate analysis of the prognosis factors for recurrence within 6 months postoperative, while the Logistic regression was used for multivariate analysis. Results:There were 30 patients (16.0%) developed recurrence within 6 months after operation, including local recurrence in 1 case, regional recurrence in 11 cases, hematogenous recurrence in 13 cases, and combined recurrence in 5 cases. Univariate analysis suggested that there was a significant difference in T staging of tumor before neoadjuvant therapy (cT), tumor regression grade, circumferential resection margin, pathological T stage (ypT) and pathological N stage (ypN) between the recurrence patients and non-recurrence patients (all P<0.05). Logistic regression analysis suggested that the cT3-4 ( OR=2.701, 95% CI: 1.161 to 6.329, P=0.021) and ypN(+)( OR=1.654, 95% CI: 1.045 to 2.591, P=0.032) were the independent prognosis factors for recurrence within 6 months. Conclusion:The combination of neoadjuvant therapy and surgery is not effective in reducing early postoperative recurrence in patients who have invaded the epineurium before treatment, and still have positive lymph nodes after neoadjuvant therapy.