Preparation of a Single-Cell Suspension from Tumor Biopsy Samples for Single-Cell RNA Sequencing.

An essential requirement for single-cell RNA sequencing in cancer is the preparation of high-quality single-cell suspensions from the tumor tissue. In this work, various methods of dissociation of tumor biopsy specimens were analyzed and developed to obtain a cell suspension with at least 80% viability. It was found that the optimal conditions for sample preparation are mechanical dissociation followed by incubation with a collagenase/hyaluronidase mixture with addition of DNAase I for 60 min. Thus, we optimize the approach for preparing single-cell suspensions from the tumor biopsy tissue for single-cell RNA sequencing.

[High-tone therapy in complex rehabilitation of patients with post-sternotomy syndrome after coronary artery bypass surgery. (Open-label, prospective, randomized controlled trial)]. / Vysokotonovaya terapiya v kompleksnoi reabilitatsii bol'nykh s poststernotomicheskim sindromom posle aortokoronarnogo shuntirovaniya. (Otkrytoe prospektivnoe randomizirovannoe kontroliruemoe issledovanie).

INTRODUCTION: At present it remains relevant to develop new rehabilitation technologies for patients with circulatory system diseases who have undergone a cardiac surgery to restore the functions of the cardiorespiratory system more quickly, improve physical and mental health, and prevent the development of the atherosclerotic process. AIM: To study the effectiveness and safety of the new rehabilitation technology for the inpatient stage of medical rehabilitation of patients with post-sternotomy syndrome after coronary artery bypass surgery (CABS) using high-tone therapy. MATERIAL AND METHODS: The study included 85 men (the average age was 56.8±2.46 years old) with post-sternotomy syndrome after CABS. They were divided into two groups by simple randomization: the first/main (42 patients) and the second/control (43 patients). The control group of the patients had a standard rehabilitation complex; the main group was additionally prescribed a high-tone therapy according to a local method. The immediate results of the treatment were assessed by the dynamics of the clinical picture, the six-minute walk test, respiratory function, echocardiography, the level of cytokines, C-reactive protein and natriuretic peptide (NT-proBNP); distant - by QOL endpoints (questionnaire MOS SF-36). RESULTS: The groups of the patients were comparable in all baseline parameters. After the course of the procedures in the main group of patients there were positive reliable (p<0.05-0.001) shifts in clinical (pain, shortness of breath, general weakness), functional (forced expiratory vital capacity, forced expiratory volume1, effusion separation) and laboratory parameters (leukocytes, interleukin-2 and 10, NT-proBNP). The intergroup analysis of long-term results registered significant (p<0.05) differences in the QOL of patients in the main group by subscales: the role of somatic problems, vitality and mental health. Compliance to the III stage of medical rehabilitation (outpatient/home) was noted with 95.2% of the patients in the first group and 93.0% in the second. CONCLUSION: The additional appointment of a high-tone therapy to the rehabilitation standard for the patients with post-sternotomy syndrome after CABS significantly improves the immediate and long-term results of the treatment (QOL) contributing to a more pronounced reverse development of inflammatory and edematous syndromes, an increase in physical activity and psychosomatic health. The absence of adverse reactions with all the patients indicates the safety of rehabilitation complexes.

[The effect of internal course intake of selenium-modified mineral water on bone mineral density after long-term diet-induced metabolic disorders in the experiment. (Randomized controlled trial)]. / Vliyanie vnutrennego kursovogo priema modifitsirovannykh selenom mineral'nykh vod na mineral'nuyu plotnost' kostnoi tkani posle dlitel'nogo diet-indutsirovannogo narusheniya metabolizma v eksperimente. (Randomizirovannoe kontroliruemoe issledovanie).

INTRODUCTION: The phenomenon of therapeutic and prophylactic effect of natural mineral waters (MW) used to restore tissue tolerance to glucose and normalize basal insulin secretion in metabolic syndrome is well known. At the same time, the use of selenium-enriched MW is promising due to its multitarget action including participation in the regulation of the protein-synthesizing function of cells. AIM: To study the effect of internal course intake of selenium-modified mineral waters of different composition on bone mineral density after long-term diet-induced metabolic disorders in the experiment. MATERIAL AND METHODS: The study was carried out on outbred male white rats that were distributed into 3 control groups (CG) and 4 main groups (MG). In CG1 14 animals received laboratory food and drinking water; in CG2 14 animals were modeled for diet-induced metabolic disorders, and they were withdrawn from the experiment the day after the cancellation of the hypercaloric diet (180 days); after the cancellation of the hypercaloric diet 180 days later 14 animals with diet-induced metabolic disorders in CG3 had a standard diet (laboratory food and drinking water) for 30 days. In the experimental groups (EG), the animals similarly to CG3 were modeled diet-induced metabolic disorders, after the cancellation of the diet they had a standard diet and watering with MW. In MG1 18 animals were given MW1 (Essentuki-4); in MG2 19 animals had a course of MW1Se (Essentuki-4 enriched with selenium); in MG3 14 animals had a course of MW2 (Slavyanovskaya); in MG4 18 animals had a course of MB2Se (Slavyanovskaya enriched with selenium). The weight (body weight) of the animals was monitored monthly. Biochemical studies, densitometry and computed microtomography were used to determine the effectiveness of MW. RESULTS: The characteristics of spongy bone tissue of femoral metaphysis of the rats of the control groups revealed disorders of varying degrees, but, despite this, the bone tissue structure of the examined vertebral bodies in animals without a hyper caloric diet (CG1) was preserved. The analysis of the study results demonstrated a significant effectiveness of selenium-enriched MW in regulating metabolic processes (p<0.05-p<0.01), bone mineral density (p<0.05), reduced bone resorption (p<0.05), that is, the characteristics of spongy bone tissue of femoral metaphysis and vertebral bodies of animals showed a clear tendency to restore bone structure. CONCLUSION: The results of this study make it possible to predict an increase in the effectiveness of medical rehabilitation of patients with somatic diseases when selenium-enriched MW is included in the programmes.

[Nonspecific inflammatory reaction after implantation of palpebral implants]. / Nespetsificheskaya vospalitel'naya reaktsiya posle implantatsii pal'pebral'nogo implantata.

PURPOSE: To identify the possible cause of an inflammatory reaction to a Russian-manufactured palpebral implant made of gold in the long term after surgery, and to determine the clinical and morphological changes in the tissues of the upper eyelid when the presence of the implant caused the inflammatory reaction. MATERIAL AND METHODS: The results of 150 operations with placement of a palpebral implant were analyzed. In 12 cases, a nonspecific inflammatory reaction was revealed within 2 to 4 weeks after the operation, in 7 cases it necessitated explantation, in 5 cases the local long-term use of an ointment with a corticosteroid made it possible to avoid extrusion and explantation. Chemical microanalysis of the palpebral implant and fragments of the removed capsule was performed using scanning electron microscopy (SEM), as well as immunohistochemical (IHC), macro- and microscopic examination of the fragments of connective tissue capsule was carried out after removal of the palpebral implant. RESULTS: The obtained data confirm the chemical purity of the implant, the absence of abnormal accumulation of metals in the tissues of the eyelid. IHC, macro- and microscopic examination of the presented fragments of the connective tissue capsule revealed signs characteristic of an inflammatory reaction to a foreign body. CONCLUSION: Further research is needed to establish the factors and predictors for the development of inflammatory reactions to a foreign body and, in particular, to gold.

[Granulomatous inflammation of the orbit as a complication of stem cells injection (case study)]. / Granulematoznoe vospalenie orbity kak oslozhnenie intraorbital'noi in"ektsii stvolovykh kletok (klinicheskoe nablyudenie).

The article present a case report of orbital granulomatous inflammation after a retrobulbar injection of allogenous stem cells. Experimental treatment resulted in an orbital tumor that required surgical excision. Lymphogranulomatous inflammation with a secondary abscess was verified by morphological and immunohistochemical analyses. This case demonstrates the possible dangerous complications of the "off-label" therapy amid the rising popularity of stem cells treatment.

[Experimental investigation of the efficacy of pirfenidone in prevention of ostium cicatricial closure after dacryocystorhinostomy]. / Eksperimental'noe obosnovanie effektivnosti pirfenidona v predotvrashchenii rubtsovogo zarashcheniya iskusstvennogo soust'ya posle dakriotsistorinostomii.

One of the main reasons of failure of dacryocystorhinostomy (DCR) is cicatricial closure of the ostium. Finding a way to prevent this outcome remains one of the leading aims of research in dacryology. The effectiveness of the most widespread methods is often considered contradictory by various researchers. Pirfenidone is a small-molecule agent that demonstrated good antifibrotic effect and low toxicity in previous in vitro research. There haven't been any in vivo studies of its intraoperative use in DCR.Purpose - to determine the in vivo efficacy of pirfenidone in prevention of ostium cicatricial closure following dacryocystorhinostomy in an animal experiment. MATERIAL AND METHODS: The study was conducted on 18 Chinchilla rabbits. They were divided into 3 groups and each animal underwent modified dacryocystorhinostomy. On the final stage of surgery rabbits of group 1 were injected 1 ml of 0.15 mg/ml pirfenidone solution, rabbits of group 2 - 0.3 mg/ml pirfenidone solution. No injections were performed in group 3. Animals were terminated on days 7 (6 rabbits), 14 (6 rabbits) and 28 (6 rabbits) following surgery. Lacrimal stoma patency was evaluated in vivo by irrigation, and morphologically postmortem. Tissue samples obtained from the stoma area were examined histologically for signs of fibrosis. RESULTS: Failure of dacryocystorhinostomy was observed in 4 out of 18 cases: all rabbits of group 3 terminated on days 14 and 28. The most pronounced morphological signs of fibrosis were also noted in group 3. No topical or systemic adverse effects of the medication were observed in groups 1 and 2. CONCLUSION: Pirfenidone demonstrated high antifibrotic efficacy and low toxicity in experimental dacrycystorhinostomy in rabbits. These results provide grounds for further research into the use of pirfenidone in dacrycystorhinostomy.

[Experimental investigation of the safety of terahertz radiation in corneal hydration assessment]. / Eksperimental'noe obosnovanie bezopasnosti primeneniya teragertsovogo izlucheniya dlya analiza izmenenii gidratatsii rogovitsy.

Application of terahertz (THz) radiation in novel non-invasive biomedical technologies has recently received considerable attention. However, experimental data about the safety of exposure to THz radiation for biological objects (including eye structures in vivo) are limited. To our knowledge, the safety of THz reflectometry (frequency range of 0.30-0.40 THz) has not been closely examined in an animal model with subsequent morphological assessment of corneal tissues. PURPOSE: To assess the safety of pulsed THz radiation with various parameters (time, power, and frequency) for the cornea in a rabbit model. MATERIAL AND METHODS: The sample for the current study consisted of 18 Chinchilla rabbits (18 eyes). Corneal imaging and epithelial cell density before and after the exposure were evaluated using confocal laser scanning microscopy (CLSM). The histological study for objective assessment of the cornea state (day 1 and day 14) was performed after experiment termination. RESULTS: Single and multiple exposures of laser radiation at a frequency below 0.1 THz and power density below 30 nW/cm2 do not cause visible structural changes in any layers of the rabbit cornea. The results obtained in the long-term period showed insignificant reversible morphological changes only within the epithelium. CONCLUSION: The described parameters of terahertz and subterahertz radiation can be considered safe for assessing changes in corneal epithelium hydration level using non-invasive methods based on THz reflectometry.

[Uterine adenosarcoma. Report of 5 cases and review of literature]. / Adenosarkoma matki. Opisanie pyati nablyudenii i obzor literatury.

Uterine adenosarcoma is an uncommon biphasic tumor with benign epithelial and malignant mesenchymal components, often presenting difficulties for morphological diagnosis. We describe 5 cases of adenosarcoma of the uterine corpus and cervix, and vaginal stump in patients aged 46-76 years. Clinical data, ultrasound results, morphological data, including immunohistochemical studies with antibodies to CD10, estrogen and progesterone receptors, desmin, smooth muscle actin, and Ki-67 are presented. Large polypoid mass of the epithelial-mesenchymal structure within the uterine cavity in women of peri - and postmenopausal age require the exclusion of malignancy of the mesenchymal component with searchig for diagnostic criteria - periglandular cuffing of the stromal cells and mitoses.

[Histological verification of autofluorescence borders of periorbital skin tumors]. / Gistologicheskaya verifikatsiya autofluorestsentnykh granits novoobrazovanii kozhi periorbital'noi oblasti.

Tumor borders are one of the most significant characteristics of any tumor, including that of the skin. PURPOSE: To compare histological borders of periorbital skin tumors with their autofluorescence borders built from the analysis of non-induced protoporphyrin IX autofluorescence. MATERIAL AND METHODS: The study group included 8 patients with skin tumors of the eyelids, periorbital region, eyebrow and zygomatic regions aged 54-88 years. The tumors varied in size from 2 to 8 mm and all displayed signs of basal cell carcinoma (BCC). At admission, all the patients underwent non-induced autofluorescence diagnosis. The images were processed with the «CancerPlot¼ program. During radio excision, the autofluorescent border of each neoplasm was marked with a surgical incision of about 5 mm long and 2 mm deep. RESULTS: Upon pathomorphological examination, solid BCC was identified in 7 cases. The remaining case was senile keratosis. All reference incisions were located in healthy tissues not farther than 1 mm from the tumor (or keratosis locus, correspondingly). CONCLUSION: By the example of facial BCC, an evident correlation was established between histological borders of the tumor and its native (non-induced) protoporphyrin IX autofluorescence.

[Modified meibography in malignant eyelid tumors of epithelial origin]. / Modifitsirovannaia meibografiia pri zlokachestvennykh novoobrazovaniiakh vek épitelial'nogo proiskhozhdeniia.

INTRODUCTION: Eyelid tumors can be diagnosed using various diagnostic techniques. However, capability of existing methods for determining the type and margins of the tumor at the preoperative stage remains insufficiently studied. PURPOSE: To improve the noncontact infrared meibography technique and determine its diagnostic capabilities for studying structural changes in meibomian glands in malignant eyelid tumors of epithelial origin. MATERIAL AND METHODS: The study included 49 people (66 eyelids), among which 21 (21 eyelids) with malignant eyelid tumors, 11 - with benign eyelid tumors (11 eyelids), and 17 (34 eyelids) comprised the control group. The mean age of patients was 64.8±1.56 years. Meibomian glands of the lower eyelids were photographed in infrared light. Receiver-operating characteristic (ROC) curves were constructed to estimate the sensitivity and specificity of the method. RESULTS: Modified meibography showed 100% sensitivity and 90% specificity in the differential diagnosis of malignant and benign eyelid tumors in 32 patients. Unlike other methods, the modified meibography technique can clarify the margins of tumor growth, which is considered when planning the resection volume. CONCLUSION: Modified meibography can indicate the margins of tumor invasion; it can be successfully used in the differential diagnostics of malignant eyelid tumors of epithelial origin and benign eyelid tumors.

[Inflammatory granuloma after intracanalicular punctal plug migration (a clinical case)]. / Vospalitel'naia granulema pri vnutrikanal'tsevoi migratsii okkliudera sleznoi tochki (sluchai iz praktiki).

At present, installation of punctal plugs (tear duct occluders) draws attention of ophthalmologists, but this method of treating dry eye syndrome (DES) is not without complications. Considering the rise of DES occurrence - the tendency anticipated to continue - as well as expansion of indications for installation of tear duct occluders, their usage can be expected to rise. The article describes a relatively rare clinical case that involved intracanalicular migration of silicone punctal plug. A female patient of 36 years old sought medical help in Research Institute of Eye Diseases (Moscow) to treat a lump in the area of lower lacrimal punctum in the left eye that was growing in size; the lump had appeared around 2 months prior to the visit. Patient's medical history read that around 2 years ago she had a silicone occluder installed in the lower lacrimal punctum of the left eye. On examination, in the area of lower lacrimal punctum, a body with a nutrient vascular pedicle deriving from lower lacrimal duct could be found. The occluder was absent in the opening of the lacrimal punctum. A revision of lower tear duct cavity was performed to remove its contents. The body filling tear duct opening was removed with forceps. Substance was then sent for histological examination. Tear duct was scraped out, the silicone occluder removed and sent to laboratory for scanning electron microscopy. The patient had no complaints 6 months after the procedure. CONCLUSION: The study showed that the forming body was granuloma resulting from aseptic inflammation. Surface of the silicone occluder in retention of lacrimal pathways remained unchanged. Described surgical tactic is suitable for treating patients with intracanalicular punctal plug migration.

[The nasal mucosa and outcomes of dacryocystorhinostomy]. / Sostoianie slizistoi obolochki polosti nosa i rezul'taty dakriotsistorinostomii.

OBJECTIVE: To investigate the impact of the nasal mucosa on the outcomes of dacryocystorhinostomy on the basis of morphologic findings. MATERIAL AND METHODS: The investigation enrolled 73 patients who had undergone endonasal endoscopic dacryocystorhinostomy. Nasal mucosal biopsies were intraoperatively taken from all the patients. The obtained samples were subjected to standard histological examination, as well as to immunohistochemical study using an anti-alpha-smooth muscle actin antibody. To determine the intensity of inflammation in the tissue sample, a chronic inflammation score was calculated. The cell elements positively stained with α-smooth muscle actin were estimated using a semi-automatic method. The patients were divided into groups in accordance with the outcome of surgical treatment after 6 months. RESULTS: An unfavorable outcome of dacryocystorhinostomy was observed in 10 (13.7%) patients. The samples obtained from the patients showed a higher chronic inflammation score (8.33%) and a larger number of the cell elements positively stained with α-smooth muscle actin (6026.38±1944.29). The correlation between the outcome of surgical intervention and the quantitative characteristics of myofibroblasts was statistically significant (p<0.05). CONCLUSION: These studies suggest that there is a direct correlation of the efficiency of endonasal endoscopic dacryocystorhinostomy with the presence and degree of chronic nasal mucosal inflammation at baseline.

[The substantiation of the application of the high-intensity pulsed magnetic field for the combined treatment of biliary sludge]. / Obosnovanie primeneniia vysokointensivnogo impul'snogo magnitnogo polia v kompleksnoi terapii biliarnogo sladzha.

Despite the considerable achievements in the field of gastroenterology, there is still high incidence of diseases of the organs of the hepatobiliary system which necessitates the development of new therapeutic techniques for their management. Nowadays, high-intensity pulsed magnetic therapy is considered to be a highly efficacious method characterized by well-pronounced and many-sided action on the processes proceeding in the organism as a new preformed factor producing neurostimulatory, vasodilatory, trophic, and hypoalgesic effects. It appears to be effective with respect to biliary sludge (BS) since it can promote depletion of stagnant contents of the gall bladder and also intensify its contractile function. AIM: The objective of the present study was to evaluate the effectiveness of the application of the high-intensity pulsed magnetic field in the combination with the intake of mineral water (MW) and rational pharmacotherapy as the components of the combined treatment of biliary sludge. METHODS: The study included 117 patients presenting with biliary sludge (BS) who were randomly allocated to three groups. Group 1 was comprised of 38 patients and served as the control group. These patients received medicamentous therapy (hymecromone - 200 mg 3 times daily during two weeks and ursodeoxycholic acid at a dose of 10-15 mg/kg a day during a period from 3 to 6 months). The second group consisted of 40 patients and served as the group of comparison. The patients of this group received medicamentous therapy in the combination with the intake of 'Sernovodskaya' hydrocarbonate-chloride-sulphate sodium mineral water (from a spring in the Chechen republic). The third group was composed of 39 patients and constituted the main study group. These patients completed a course of therapy with the use of the high-intensity pulsed magnetic field applied to the gall bladder region. Each patient underwent 10 sessions of magnetic therapy after the completion of the treatment with hymecromone and 'Sernovodskaya' mineral water. The treatment was carried out with simultaneous monitoring dynamics of the clinical symptoms of biliary sludge, the ultrasound examination of the abdominal organs with the evaluation of the contractile function of the gall bladder, the level of bilirubin in the cystic bile, bile acids, cholesterol, and cholate-cholesteric coefficient. RESULTS: The combined treatment of the patients suffering from BS including the application of the high-intensity pulsed magnetic field alleviated the clinical symptoms of the disease much faster than medicamentous therapy combined with the intake of 'Sernovodskaya' mineral water. Moreover, it promoted the restoration of the motor activity of the gall bladder and accelerated the evacuation of the hyperechoic particles. CONCLUSION: The course of the combined treatment of the patients presenting with biliary sludge including the application of the high-intensity pulsed magnetic field in the combination with the intake of mineral water (MW) and rational pharmacotherapy significantly relieved abdominal pains, alleviated the symptoms of biliary dyspepsia, and improved the motor function of the biliary tract. These therapeutic effects persisted as long as 3 months. The results of the present study give evidence of the necessity of prescription of an optimum dose of ursodeoxycholic acid to the patients suffering from biliary sludge.

[The role of the combined balneotherapeutic treatment as the 'add-back therapy' against the background of the anti-hormonal effects of the agonists of gonadotropin-releasing hormone in the women suffering from endometriosis genitalis externa]. / Kompleksnoe bal'neolechenie v kachestve terapii prikrytiia na fone antigormonal'nogo vozdeistviia agonistami gonadotropin-rilizing gormona u zhenshchin s naruzhnym éndometriozom.

BACKGROUND: The use of agonists of gonadotropin-releasing hormone (GnRH) for the rehabilitation treatment of the patients presenting with endometriosis genitalis externa is known to significantly enhance the risk of development of hypoestrogenism. The so-called 'add-back-therapy' is carried out as a preventive measure to eliminate hypoestrogenism caused by the intake of GnRH agonists without detriment to the effectiveness of anti-hormonal therapy. AIM: The objective of the present study was to estimate the effectiveness of the new method ('add-back hormone replacement therapy') in comparison with anti-hormonal therapy with the use of GRH agonists at the stage of the early postoperative medical rehabilitation based on the drinking of mineral waters (MW) and radon therapy (RT) in the patients who had undergone the laparoscopic interventions for the treatment of endometrioid heterotopies of the ovaries (1-3 months post-operatively). MATERIAL AND METHODS: To evaluate the effectiveness of the spa and health resort-based rehabilitation measures, the long-term effects of GnRH agonists have been studied in 2 groups of the patients formed by the of simple randomization method. The main group (MG) was comprised of 45 women received medical rehabilitation under conditions of a spa and health resort setting with the application of medium-salt water from the Pyatigorsk spring and radon therapy in the combination with triptorelin. The group of comparison (CG) was composed of 44 women given the treatment with the use of triptorelin under the out-patient conditions. The assessment of the results of the study included: the estimation of the intensity of pelvic pain syndrome, indicators of the hormonal status and the quality of life (QL), ultrasonic examination of the pelvic organs, and the occurrence of adverse effects of triptorelin. The duration of the study was 3 years (2014-2016). RESULTS: The analysis of the long-term results of the rehabilitative treatment has demonstrated that the rehabilitation activities under the spa and health resort conditions within 1-3 months after the surgical intervention including the application of MW and RT in the combination with an agonist of gonadotropin-releasing hormone (triptorelin) greatly contributed to the stability of the results of the treatment: specifically, only 2.5% of the patients suffered the recurrence of cystic ovarian endometriosis whereas 24.5% (p<0.001) reported the restoration of the reproductive function. The patients comprising the control group experienced the recurrence of the pathologic process in 7.5% of the cases and the restoration of the reproductive function in 15.8% of the cases (p<0.001). Simultaneously, the quality of life in the patients in the main group improved 3.4 times on the average in comparison with the initial level (p<0.001). The 2.2-fold improvement of the quality of life was documented in the control patients (p<0.001). At the same time, the frequency of adverse reactions to the treatment with triptorelin in the control patients was higher than in the main group including pain in the epigastric area, headache, irritability, hyperhidrosis, and instability of arterial blood pressure (ABP) that significantly deteriorated the life quality in these women. CONCLUSION: The comparative analysis of the results of the follow-up observations has demonstrated that the application of drinking mineral water and radon therapy for the treatment of the women suffering from endometriosis genitalis externa can be recommended as 'add-back therapy' against the background of anti-hormonal effects of agonists of gonadotropin-releasing hormone.

[Surgical treatment of purulent corneal ulcers in eyes with neurotrophic keratitis and paralytic lagophthalmos]. / Lechebnaia keratoplastika pri gnoinykh iazvakh rogovitsy, razvivshikhsia na fone neirotroficheskogo keratita i paraliticheskogo lagoftal'ma.

AIM: to develop a surgical approach to the treatment of purulent corneal ulcer in eyes with neurotrophic keratitis with lagophthalmos after auditory neuroma removal or stroke. MATERIAL AND METHODS: A total of 12 patients (13 eyes) have been managed. All received combined surgery that included simultaneous keratoplasty, autoconjunctivoplasty, and permanent partial tarsorrhaphy. RESULTS: Ocular integrity was preserved in all cases, and also vision - in half of them. In 29% of cases, visual acuity increased from 0.09±0.05 up tо 0.21±0.13. Lagophthalmos decreased from 5.86±1.35 mm down to 3.01±0.75 mm. CONCLUSION: Patients with intracranial lesions complicated by neurotrophic keratitis and lagophthalmos often develop progressive purulent ulcers of the cornea. One-step keratoplasty, autoconjunctivoplasty, and permanent partial tarsorrhaphy is an effective treatment that can help preserve visual function in such eyes.

[Combined YAG-laser capsulotomy in pseudophakic patients with anterior capsular contraction syndrome]. / Tekhnologiia kombinirovannoi lazernoi kapsulotomii pri perednekapsuliarnom kontraktsionnom sindrome v artifakichnykh glazakh.

Progressive metaplastic fibrosis of the anterior capsulorhexis opening is a frequent complication of the postoperative period. There is a widely practiced method of anterior capsular contraction syndrome (ACCS) correction through radial anterior laser capsulotomy. Despite many advantages, it can be complicated by unpredictable anterior capsule tearing and intraocular lens (IOL) dislocation into the vitreous Body, which justifies the search for new technical solutions. AIM: to develop a safe technique of combined laser anterior capsulotomy (LAC) in patients with ACCS. MATERIAL AND METHODS: The study included 19 patients (20 eyes) with ACCS. The suggested LAC technique involved two differently directed YAG-laser cuts that could be regarded as a combination of radial anterior capsulotomy and anterior peripheral linear capsulotomy. With the cuts located perpendicularly to each other, the distal end of each radial cut was connected to the middle of each longitudinal cut. RESULTS: The suggested technique allows an increase in the anterior capsulorhexis diameter up to more than twice the pre-laser size. The difference between the average pre- and post-laser capsulorhexis diameters was statistically significant (р<0.0001). Neither case developed an unpredictable anterior capsule tear. CONCLUSION: The safety of the new technique is ensured by preliminary longitudinal cuts that create a barrier against unpredictable radial tears in the capsular bag during radial capsulotomy.

[Gold weight implants for lagophthalmos correction in chronic facial nerve paralysis (late results)].

UNLABELLED: Upper eyelid weigh gold implant is the widely accepted standard for the treatment of paralytic lagophthalmos (PL). AIM: To evaluate late outcomes of PL correction with chain gold implants. MATERIAL AND METHODS: Chain gold implants were inserted in the upper eyelids of 70 patients with lagophthalmos due to chronic facial paralysis. A comprehensive ophthalmic examination was performed prior to surgery and then at months 1, 3, 6, 12, 24, and 36. RESULTS: The results obtained prove the method highly effective. None of the patients developed any severe complications. Cases of implant removal were few. Some of the studied biometric parameters decreased significantly after surgery, while the upper eyelid excursion increased. The implant had no effect on the inner surface of the cornea and its peripheral thickness. CONCLUSION: Efficacy of the proposed eyelid implant has been convincingly demonstrated; late complications have been analyzed.

[Morphological assessment of lens capsule after different techniques of cataract extraction]. / Morfologicheskaya otsenka izmenenii kapsuly khrustalika posle razlichnykh sposobov ekstraktsii katarakty.

AIM: to evaluate morphological changes in the posterior lens capsule (LC) after different techniques of cataract extraction. MATERIAL AND METHODS: Eight pseudophakic human autopsy eyes were divided into two groups by the technique used for cataract extraction. Group 1 included 4 eyes following extracapsular cataract extraction (ECCE) and rigid IOL implantation. Group 2 included 4 eyes after minimally invasive cataract surgery (MICS) that involves the use of ultrasound energy. Semithin LC sections were polychromatically stained, visually examined, and imaged for further analysis. RESULTS: There were certain common features in clinical and morphological appearances of lens capsules from either group, including fibrous metaplasia of epithelial cells and/or pseudoregenerative Elschnig-Adamuk pearls on their inner surfaces, more significant in the ECCE group. MICS group, however, demonstrated many distinct and previously undescribed changes. In particular, post-MICS LCs looked swollen, flabby, and even amorphous, their architectonics were disturbed and fibers separated forming micro slit-like spaces, which could be a result of acoustic cavitation caused by ultrasound exposure. We have named this morphological type of secondary cataract «hyaloid-like¼. CONCLUSION: Clinical and morphological type of postoperative LC opacification as well as its severity depends, to a certain extent, on the technique used for cataract extraction. Hyaloid-like opacifications are typical of minimally invasive surgery involving the use of ultrasound and often show no 'classic' changes (fibroproliferative and pseudoregenerative). The latter is explained by the fact that the posterior LC, which has lost its biomechanical properties, can no longer be a substrate for proliferating cells.

[SEM visualization of corneal epithelium through lanthanoid staining based on Ca/Nd isomorphous substitution in Ca-dependent molecular systems]. / Vizualizatsiya struktury epiteliya rogovitsy metodom skaniruyushchei elektronnoi mikroskopii s lantanoidnym kontrastirovaniem na osnove Ca/Nd izomorfnogo zameshcheniya v Sa-zavisimykh molekulyarnykh sistemakh.

Cumulative biomicroscopic evidence is usually sufficient for the diagnosis of recurrent corneal erosion or bullous keratopathy, however, exploration of the disease pathogenesis requires subcellular-level visualization of corneal structure. In the current study, lanthanoid staining and scanning electron microscopy were employed to visualize quite a number of structures responsible for epithelium organization. In particular, the study proves possible the use of Ca/Nd isomorphous substitution at Ca2+ sites of cytoadherence proteins for visualization of corresponding cellular structures. AIM: To assess the value of information provided by scanning electron microscopy of corneal epithelium that involves lanthanoid staining based on the Ca/Nd isomorphous substitution in Ca-dependent molecular systems. MATERIAL AND METHODS: Anterior corneal epithelial scrapes were obtained from patients with recurrent corneal erosion or bullous keratopathy and cadaver eyes with no signs of any ophthalmic disease. Samples were then studied under a scanning electron microscope (Zeiss EVO LS10, BSE, EP - 79 Pa, 20-28 kV, Ln-staining with the BioREE assay kit). RESULTS: In all cases, lanthanoid staining of biopsy material provided high-contrast SEM images with well-recognizable structural and ultrastructural elements associated with Ca2+ sites of cytoadherence proteins. CONCLUSION: Lanthanoid staining of biopsy material and subsequent SEM enabled detailed visualization of structural features of the corneal epithelium in various pathologies. Due to the Ca/Nd isomorphism we were able to evaluate structural position of the majority of protein molecules engaged in Ca-dependant processes and, consequently, in cytoadherence. Basing on the neodymium distribution within the basal membrane, we have described local effects of different substances on the lamina densa in the projection of basal layer cell borders that occur after unidirectional ultrafiltration. The results confirm the failure of the junctional adhesion complex in recurrent corneal erosion.

[Morphological analysis of reparative process at osteotomy site after endoscopic endonasal dacryocystorhinostomy]. / Morfologicheskii analiz protsessovreparatsii v oblasti dakriostomy posle endoskopicheskoi endonazal'noi dakriotsistorinostomii.

Ostium closure due to local reparative processes is the most common cause of recurrence after endoscopic endonasal dacryocystorhinostomy (DCR), but as yet poorly studied. AIM: To perform morphological study of reparative processes at osteotomy site after endoscopic endonasal DCR. MATERIAL AND METHODS: The study included 18 patients (18 women) aged 62.59±10.07 years with nasolacrimal duct obstruction, who underwent endoscopic endonasal DCR (P.J. Wormald modification). Osteotomy site biopsies were taken on days 2, 5, 7, 10, 14, 21, 28, and 60 after the surgery. All the samples were stained with haematoxylin and eosin. For further details on cellular composition of inflammatory infiltrate, 54 samples were also processed into semi-thin sections. The slides were then viewed under Leica DM-2500 (Leica, Germany) photomicroscope. Leica DFC320 digital camera and ImageScope Color software were used for image acquisition and analysis. RESULTS: Morphological examination revealed the predominance of inflammatory cells in biopsy material on day 1 after DCR. On day 14, the number of cells would usually be reduced indicating the completion of the exudative phase of inflammation. Since then, activated fibroblasts prevailed over the rest of cellular elements. Twenty eight days after DCR, proliferating fibroblasts and collagen fibers were present. Sixty days after the surgery, fibrillar component prevailed over cells and showed compaction. CONCLUSION: Our results demonstrate that the reparation process nears completion before day 60 after the surgery. A series of morphological examinations of biopsy material from osteotomy sites has revealed individual features of reparation in nasal and lacrimal sac mucosa after endoscopic endonasal DCR.